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Park K, Shin M, Natasha A, Kim J, Noh J, Kim SG, Kim B, Park J, Seo YR, Cho HK, Byun KS, Kim JH, Lee YS, Shim JO, Kim WK, Song JW. Novel human coronavirus in an infant patient with pneumonia, Republic of Korea. Emerg Microbes Infect 2025; 14:2466705. [PMID: 39945663 PMCID: PMC11849027 DOI: 10.1080/22221751.2025.2466705] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2024] [Revised: 02/06/2025] [Accepted: 02/09/2025] [Indexed: 02/22/2025]
Abstract
Coronaviruses (CoVs) pose a significant threat to public health, causing a wide spectrum of clinical manifestations and outcomes. Beyond precipitating global outbreaks, Human CoVs (HCoVs) are frequently found among patients with respiratory infections. To date, limited attention has been directed towards alphacoronaviruses due to their low prevalence and fatality rates. Nasal swab and serum samples were collected from a paediatric patient, and an epidemiological survey was conducted. Retrospective surveillance investigated the molecular prevalence of CoV in 880 rodents collected in the Republic of Korea (ROK) from 2018 to 2022. Next-generation sequencing (NGS) and phylogenetic analyses characterized the novel HCoV and closely related CoVs harboured by Apodemus spp. On 15 December 2022, a 103-day-old infant was admitted with fever, cough, sputum production, and rhinorrhea, diagnosed with human parainfluenza virus 1 (HPIV-1) and rhinovirus co-infection. Elevated AST/ALT levels indicated transient liver dysfunction on the fourth day of hospitalization. Metagenomic NGS (mNGS) identified a novel HCoV in nasal swab and serum samples. Retrospective rodent surveillance and phylogenetic analyses showed the novel HCoV was closely related to alphacoronaviruses carried by Apodemus spp. in the ROK and China. This case highlights the potential of mNGS to identify emerging pathogens and raises awareness of possible extra-respiratory manifestations, such as transient liver dysfunction, associated with novel HCoVs. While the liver injury in this case may be attributable to the novel HCoV, further research is necessary to elucidate its clinical significance, epidemiological prevalence, and zoonotic origins.
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Affiliation(s)
- Kyungmin Park
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
- Institute for Viral Diseases, Korea University College of Medicine, Seoul, Republic of Korea
| | - Minsoo Shin
- Department of Paediatrics, Korea University Ansan Hospital, Ansan, Republic of Korea
| | - Augustine Natasha
- Department of Microbiology, College of Medicine, Hallym University, Chuncheon, Republic of Korea
| | - Jongwoo Kim
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
- BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul, Republic of Korea
| | - Juyoung Noh
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
| | - Seong-Gyu Kim
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
- BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul, Republic of Korea
| | - Bohyeon Kim
- Department of Microbiology, College of Medicine, Hallym University, Chuncheon, Republic of Korea
| | - Jieun Park
- Department of Microbiology, College of Medicine, Hallym University, Chuncheon, Republic of Korea
| | - Ye-rin Seo
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
| | - Hee-Kyung Cho
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
- BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul, Republic of Korea
| | - Kwan Soo Byun
- Department of Internal Medicine, Division of Gastroenterology and Hepatology, Korea University Medical Center, Seoul, Republic of Korea
| | - Ji Hoon Kim
- Department of Internal Medicine, Division of Gastroenterology and Hepatology, Korea University Medical Center, Seoul, Republic of Korea
| | - Young-Sun Lee
- Department of Internal Medicine, Division of Gastroenterology and Hepatology, Korea University Medical Center, Seoul, Republic of Korea
| | - Jung Ok Shim
- Department of Paediatrics, Korea University College of Medicine, Seoul, Republic of Korea
| | - Won-Keun Kim
- Department of Microbiology, College of Medicine, Hallym University, Chuncheon, Republic of Korea
- Institute of Medical Research, College of Medicine, Hallym University, Chuncheon, Republic of Korea
| | - Jin-Won Song
- Department of Microbiology, Korea University College of Medicine, Seoul, Republic of Korea
- BK21 Graduate Program, Department of Biomedical Sciences, Korea University College of Medicine, Seoul, Republic of Korea
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Pini L, Giordani J, Levi G, Guerini M, Piva S, Peli E, Violini M, Piras S, El Masri Y, Pini A, Visca D, Assanelli D, Muiesan ML, Latronico N, Tantucci C, on behalf of the LOTO Investigators Working Group. Long-term alveolar-capillary diffusion impairments after severe SARS-CoV-2 pneumonia. Ann Med 2025; 57:2483383. [PMID: 40152750 PMCID: PMC11956098 DOI: 10.1080/07853890.2025.2483383] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/29/2024] [Revised: 03/06/2025] [Accepted: 03/10/2025] [Indexed: 03/29/2025] Open
Abstract
BACKGROUND Persistent respiratory symptoms and impaired gas exchange are common in patients recovering from COVID-19 pneumonia. The Lung Diffusing Capacity for Carbon Monoxide (DLCO) and Carbon Monoxide Transfer Coefficient (KCO) do not adequately distinguish alveolar membrane dysfunction from vascular abnormalities. This study aimed to characterize persistent diffusion impairment in post-ICU patients with prior SARS-CoV-2 pneumonia and reduced DLCO. METHODS After hospital discharge, patients underwent spirometry, DLCO measurement, and a 6-minute walking test every six months. If DLCO remained impaired at 18-24 months, a combined Lung Diffusing Capacity for Nitric Oxide (DLNO) and DLCO assessment was performed to differentiate alveolar-capillary membrane (DmCO) and pulmonary capillary blood volume (Vc) alterations. RESULTS Among 20 patients with persistent DLCO reduction, 3 had an obstructive ventilatory pattern, 6 had restriction, and 12 had low KCO. In restrictive cases, KCO was reduced but remained within normal limits without compensation. The DLNO/DLCO ratio exceeded 113.5% predicted in all patients. DmCO was impaired in 7 patients, while Vc was reduced in 16. CONCLUSION Both DLCO determinants were affected, with vascular impairment predominating. Vc reduction was present in most patients, with mean values below the lower limit of normality, whereas DmCO was less affected and often normal. The elevated DLNO/DLCO ratio suggests that persistent DLCO reduction is primarily driven by prolonged pulmonary capillary circulation dysfunction rather than alveolar membrane alterations, highlighting the vascular component as the primary site of long-term impairment.
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Affiliation(s)
- Laura Pini
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
- Respiratory Physiopathology Unit, ASST – Spedali Civili di Brescia, Brescia, Italy
| | - Jordan Giordani
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
| | - Guido Levi
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
- Pulmonology Department, ASST – Spedali Civili di Brescia, Brescia, Italy
| | - Michele Guerini
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
| | - Simone Piva
- Department of Anesthesia, Critical Care and Emergency, ASST Spedali Civili University Hospital, Brescia, Italy
- Department of Medical and Surgical Specialties, Radiological Sciences and Public Health, University of Brescia, Brescia, Italy
| | - Elena Peli
- Department of Anesthesia, Critical Care and Emergency, ASST Spedali Civili University Hospital, Brescia, Italy
| | - Manuela Violini
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
| | - Stefano Piras
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
| | - Yehia El Masri
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
| | - Alessandro Pini
- Department of Emergency, Anaesthesiological and Resuscitation Sciences, University Cattolica Sacro Cuore, Rome, Italy
| | - Dina Visca
- Department of Medicine and Surgery, University of Insubria, Varese, Italy
- Department of Medicine and Cardiopulmonary Rehabilitation, Istituti Clinici Scientifici Maugeri IRCCS, Tradate, Italy
| | - Deodato Assanelli
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
- Internal Medicine Unit, ASST Spedali Civili di Brescia, Brescia, Italy
| | - Maria Lorenza Muiesan
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
- Internal Medicine Unit, ASST Spedali Civili di Brescia, Brescia, Italy
| | - Nicola Latronico
- Department of Anesthesia, Critical Care and Emergency, ASST Spedali Civili University Hospital, Brescia, Italy
- Department of Medical and Surgical Specialties, Radiological Sciences and Public Health, University of Brescia, Brescia, Italy
| | - Claudio Tantucci
- Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy
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Cao L, Shi S, Zhang C, Zhao C. A phycobiliprotein-based reporter assay for the evaluation of SARS-CoV-2 main protease activity. Virology 2025; 608:110540. [PMID: 40233445 DOI: 10.1016/j.virol.2025.110540] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/12/2025] [Revised: 04/08/2025] [Accepted: 04/11/2025] [Indexed: 04/17/2025]
Abstract
SARS-CoV-2 Mpro is crucial for viral replication and transcription and is highly conserved. Therefore, it is an ideal target for developing broad-spectrum antiviral drugs. To address resistance to existing drugs caused by mutations, a simple and sensitive method for detecting the activity of Mpro is needed. Considering the excellent fluorescence properties of phycobiliproteins, this study developed a phycobiliprotein-based reporter assay to evaluate Mpro activity. An engineered lyase was generated by inserting the Mpro recognition sequence between the phycobiliprotein lyases CpcF and CpcE. To ensure that the binding of CpcE and CpcF depended on the linker, a series of truncated forms were constructed. Among them, the activity of CpcE/F-10 was significantly reduced in the presence of Mpro; however, both genetic and chemical inhibition of Mpro activity reversed these results. These data indicated that the fluorescence of phycobiliproteins was negatively correlated with Mpro activity. The reporter assay developed here will contribute to determining the impact of Mpro mutations and screening for new inhibitors.
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Affiliation(s)
- Le Cao
- College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China; College of Artificial Intelligence, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China
| | - Shuyuan Shi
- College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China; College of Artificial Intelligence, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China
| | - Chaofeng Zhang
- College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China.
| | - Cheng Zhao
- College of Chemistry and Chemical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China.
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Yasmin S, Ansari MY. A detailed examination of coronavirus disease 2019 (COVID-19): Covering past and future perspectives. Microb Pathog 2025; 203:107398. [PMID: 39986548 DOI: 10.1016/j.micpath.2025.107398] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2024] [Revised: 01/07/2025] [Accepted: 02/18/2025] [Indexed: 02/24/2025]
Abstract
The COVID-19 disease has spread rapidly across the world within just six months, affecting 169 million people and causing 3.5 million deaths globally (2021). The most affected countries include the USA, Brazil, India, and several European countries such as the UK and Russia. Healthcare professionals face new challenges in finding better ways to manage patients and save lives. In this regard, more comprehensive research is needed, including genomic and proteomic studies, personalized medicines and the design of suitable treatments. However, finding novel molecular entities (NME) using a standard or de novo strategy to drug development is a time-consuming and costly process. Another alternate strategy is discovering new therapeutic uses for old/existing/available medications, known as drug repurposing. There are a variety of computational repurposing methodologies, and some of them have been used to counter the coronavirus disease pandemic of 2019 (COVID-19). This review article compiles recently published data on the origin, transmission, pathogenesis, diagnosis, and management of the coronavirus by drug repurposing and vaccine development approach. We have attempted to screen probable drugs in clinical trials by using literature survey. This systematic review aims to create priorities for future research of drugs repurposed and vaccine development for COVID-19.
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Affiliation(s)
- Sabina Yasmin
- Department of Pharmaceutical Chemistry, College of Pharmacy, King Khalid University, Abha, Saudi Arabia.
| | - Mohammad Yousuf Ansari
- MM College of Pharmacy, Maharishi Markandeshwar (Deemed to be University), Mullana, Ambala, Haryana, 133207, India; Ibne Seena College of Pharmacy, Azmi Vidya Nagri Anjhi Shahabad, Hardoi, Uttar Pradesh (U.P.) 241124, India.
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Lee CM, Kim AS, Kim M, Jeong JW, Jo S, Hwang N, Fang S. Dynamics of T Cell-Mediated Immune Signaling Network During Pathogenesis of Chronic Obstructive Pulmonary Disease. Yonsei Med J 2025; 66:354-365. [PMID: 40414827 DOI: 10.3349/ymj.2024.0227] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/02/2024] [Revised: 11/05/2024] [Accepted: 11/06/2024] [Indexed: 05/27/2025] Open
Abstract
PURPOSE Chronic obstructive pulmonary disease (COPD) is characterized by alveolar destruction and increased inflammation, leading to respiratory symptoms. This study aimed to identify the traits for COPD progression from mild to severe stages. Additionally, we explored the correlation between coronavirus disease-2019 (COVID-19) and COPD to uncover overlapping respiratory patterns. MATERIALS AND METHODS Bulk RNA sequencing was conducted on data from 43 healthy individuals and 39 COPD patients across one dataset (GSE239897) to distinguish COPD characteristics. Single-cell RNA analysis was then performed on samples from seven mild patients, seven moderate patients, and three severe patients from three datasets (GSE167295, GSE173896, and GSE227691) to analyze disease progression. Finally, single-nuclei RNA analysis was applied to data from seven healthy individuals and 20 COVID-19 patients from one dataset (GSE171524) to compare the two conditions. RESULTS Bulk RNA sequencing revealed enhanced inflammatory pathways in COPD patients, indicating increased inflammation. Single-cell RNA sequencing showed a stronger inflammatory response from mild to moderate COPD with a decrease from moderate to severe stages. COVID-19 displayed similar biological patterns to moderate COPD, suggesting that stage-specific COPD analysis could enhance COVID-19 management. CONCLUSION The analysis found that immune responses increased from mild to moderate stages but declined in severe cases, marked by reduced pulmonary T cell activation. The overlap between moderate COPD and COVID-19 suggests shared therapeutic strategies, warranting further investigation.
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Affiliation(s)
- Chae Min Lee
- Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Korea
- Department of Biomedical Sciences, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
| | - Andrew Sehoon Kim
- Department of Neuroscience, Vanderbilt University College of Arts and Science, Vanderbilt University, Nashville, USA
| | - Minki Kim
- Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Korea
- Department of Biomedical Sciences, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
| | - Jae Woong Jeong
- Department of Medicine, Yonsei University College of Medicine, Seoul, Korea
| | - Sugyeong Jo
- Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Korea
- Department of Biomedical Sciences, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
| | - Nahee Hwang
- Department of Biomedical Sciences, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
| | - Sungsoon Fang
- Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Korea
- Department of Biomedical Sciences, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
- Chronic Intractable Disease for Systems Medicine Research Center, Yonsei University College of Medicine, Seoul, Korea.
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Apoorva, Kumar A, Chakrabarti SS, Singh SK. SARS-CoV-2 accessory proteins ORF3a and ORF6 alter the miRNome of human lung epithelial cells. Mol Biol Rep 2025; 52:494. [PMID: 40402297 DOI: 10.1007/s11033-025-10596-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/13/2025] [Accepted: 05/11/2025] [Indexed: 05/23/2025]
Abstract
BACKGROUND Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of COVID-19. The accessory proteins of SARS-CoV-2 have been reported to attune host immune responses and viral pathogenicity. We have studied the effect of SARS-CoV-2 accessory proteins ORF3a and ORF6 on the expression pattern of miRNAs and their impact on cell signaling pathways in human lung epithelial cells. METHODS AND RESULTS The miRNA expression profiling of human lung epithelial cells revealed a subset of 14 and 19 differentially expressed miRNAs (DEMs) in response to SARS-CoV-2 ORF3a and ORF6, respectively. Target prediction tools and subsequent bioinformatic analysis revealed the involvement of DEMs in key signaling pathways like PI3K/AKT, TNF, MAPK, TGF-β, and NF-κB, as a bystander effect of SARS-CoV-2 ORF3a and ORF6. The target genes were validated using real-time PCR and immunoblotting techniques. The results demonstrate that SARS-CoV-2 ORF3a and ORF6 exploit host cellular miRNAs such as hsa-miR-101-3p, hsa-miR-4455, hsa-miR-10b-5p, hsa-miR-940, and hsa-miR-4483, etc. to modulate the key cellular signaling pathways like NF-κB, TGF-β, Ras, IL-17, MAPK, and TNF signaling pathways. CONCLUSIONS The present study demonstrates that SARS-CoV-2 ORF3a and ORF6 modulate the miRNA expression pattern in human lung epithelial cells. ORF3a exploits miRNAs to trigger a pro-inflammatory response, while ORF6 antagonizes IFN signaling via miRNA dysregulations to help SARS-CoV-2 in evading the host's immune response.
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Affiliation(s)
- Apoorva
- Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India
| | - Atul Kumar
- Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India
| | - Sankha Shubhra Chakrabarti
- Department of Geriatric Medicine, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India
| | - Sunit Kumar Singh
- Molecular Biology Unit, Faculty of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India.
- Dr. B R Ambedkar Center for Biomedical Research (ACBR), University of Delhi (North Campus), Delhi, 110007, India.
- Delhi School of Public Health (DSPH), University of Delhi (North Campus), Delhi, 110007, India.
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Lin HF, Jiang RD, Qin RX, Yao B, Zeng WT, Gao Y, Shi AM, Li JM, Liu MQ. Characterization of a SARS-CoV-2 Infection Model in Golden Hamsters with Diabetes Mellitus. Virol Sin 2025:S1995-820X(25)00059-8. [PMID: 40389095 DOI: 10.1016/j.virs.2025.05.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2024] [Accepted: 05/12/2025] [Indexed: 05/21/2025] Open
Abstract
Being widespread across the globe, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) keeps evolving and generating new variants and continuously poses threat to public health, especially to the population with chronic comorbidities. Diabetes mellitus is one of high-risk factors for severe outcome of coronavirus disease 2019 (COVID-19). Establishment of animal models that parallel the clinical and pathological features of COVID-19 complicated with diabetes is thus highly essential. Here, in this study, we constructed leptin receptor gene knockout hamsters with the phenotype of diabetes mellitus (db/db), and revealed that the diabetic hamsters were more susceptible to SARS-CoV-2 and its variants than wild-type hamsters. SARS-CoV-2 and its variants induced a stronger immune cytokine response in the lungs of diabetic hamsters than in wild-type hamsters. Comparative histopathology analyses also showed that infection of SARS-CoV-2 and the variants caused more severe lung tissue injury in diabetic hamsters, and may induce serious complications such as diabetic kidney disease and cardiac lesions. Our findings demonstrated that despite the decreased respiratory pathogenicity, the SARS-CoV-2 variants were still capable of impairing other organs such as kidney and heart in diabetic hamsters, suggesting that the risk of evolving SARS-CoV-2 variants to diabetic patients should never be neglected. This hamster model may help better understand the pathogenesis mechanism of severe COVID-19 in patients with diabetes. It will also aid in development and testing of effective therapeutics and prophylactic treatments against SARS-CoV-2 variants among these high-risk populations.
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Affiliation(s)
- Hao-Feng Lin
- The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Laboratory Clinical Base, State Key Laboratory of Respiratory Disease, Guangzhou Medical University, Guangzhou 510120, China
| | - Ren-Di Jiang
- State Key Laboratory of Genetic Engineering, Greater Bay Area Institute of Precision Medicine (Guangzhou), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200433, China
| | - Rui-Xin Qin
- State Key Laboratory of Reproductive Medicine and Offspring Health, Jiangsu Laboratory Animal Center, Jiangsu Animal Experimental Center of Medicine and Pharmacy, Department of Cell Biology, Animal Core facility, Key Laboratory of Model Animal, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, National Vaccine Innovation Platform, Nanjing Medical University, Nanjing 211166, China
| | - Bing Yao
- Jinling Hospital Department Reproductive Medical Center, Nanjing Medical University, Nanjing 211166, China
| | - Wen-Tao Zeng
- State Key Laboratory of Reproductive Medicine and Offspring Health, Jiangsu Laboratory Animal Center, Jiangsu Animal Experimental Center of Medicine and Pharmacy, Department of Cell Biology, Animal Core facility, Key Laboratory of Model Animal, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, National Vaccine Innovation Platform, Nanjing Medical University, Nanjing 211166, China
| | - Yun Gao
- State Key Laboratory of Reproductive Medicine and Offspring Health, Jiangsu Laboratory Animal Center, Jiangsu Animal Experimental Center of Medicine and Pharmacy, Department of Cell Biology, Animal Core facility, Key Laboratory of Model Animal, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, National Vaccine Innovation Platform, Nanjing Medical University, Nanjing 211166, China.
| | - Ai-Min Shi
- State Key Laboratory of Reproductive Medicine and Offspring Health, Jiangsu Laboratory Animal Center, Jiangsu Animal Experimental Center of Medicine and Pharmacy, Department of Cell Biology, Animal Core facility, Key Laboratory of Model Animal, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, National Vaccine Innovation Platform, Nanjing Medical University, Nanjing 211166, China.
| | - Jian-Min Li
- State Key Laboratory of Reproductive Medicine and Offspring Health, Jiangsu Laboratory Animal Center, Jiangsu Animal Experimental Center of Medicine and Pharmacy, Department of Cell Biology, Animal Core facility, Key Laboratory of Model Animal, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, National Vaccine Innovation Platform, Nanjing Medical University, Nanjing 211166, China.
| | - Mei-Qin Liu
- The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Laboratory Clinical Base, State Key Laboratory of Respiratory Disease, Guangzhou Medical University, Guangzhou 510120, China.
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8
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Gen R, Addetia A, Asarnow D, Park YJ, Quispe J, Chan MC, Brown JT, Lee J, Campbell MG, Lapointe CP, Veesler D. SARS-CoV-2 nsp1 mediates broad inhibition of translation in mammals. Cell Rep 2025; 44:115696. [PMID: 40359110 DOI: 10.1016/j.celrep.2025.115696] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2025] [Revised: 03/13/2025] [Accepted: 04/23/2025] [Indexed: 05/15/2025] Open
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) non-structural protein 1 (nsp1) promotes innate immune evasion by inhibiting host translation in human cells. However, the role of nsp1 in other host species remains elusive, especially in bats-natural reservoirs of sarbecoviruses with a markedly different innate immune system than humans. We reveal that nsp1 potently inhibits translation in Rhinolophus lepidus bat cells, which belong to the same genus as known sarbecovirus reservoir hosts. We determined a cryoelectron microscopy structure of nsp1 bound to the R. lepidus 40S ribosomal subunit, showing that it blocks the mRNA entry channel by targeting a highly conserved site among mammals. Accordingly, we found that nsp1 blocked protein translation in mammalian cells from several species, underscoring its broadly inhibitory activity and conserved role in numerous SARS-CoV-2 hosts. Our findings illuminate the arms race between coronaviruses and mammalian host immunity, providing a foundation for understanding the determinants of viral maintenance in bat hosts and spillover.
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Affiliation(s)
- Risako Gen
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Amin Addetia
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Daniel Asarnow
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Young-Jun Park
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Joel Quispe
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Matthew C Chan
- Division of Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA, USA
| | - Jack T Brown
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Jimin Lee
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA
| | - Melody G Campbell
- Division of Basic Sciences, Fred Hutchinson Cancer Center, Seattle, WA, USA
| | | | - David Veesler
- Department of Biochemistry, University of Washington, Seattle, WA 98195, USA; Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
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9
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Fernández-Vilas E, Coca JR, Labora González JJ, Iglesias Carrera M. The Sociology of Suicide After COVID-19: Assessment of the Spanish Case. Behav Sci (Basel) 2025; 15:606. [PMID: 40426384 DOI: 10.3390/bs15050606] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2025] [Revised: 04/22/2025] [Accepted: 04/24/2025] [Indexed: 05/29/2025] Open
Abstract
The phenomenon of suicide has become a significant global concern, claiming over 800,000 lives annually and resulting in millions of suicide attempts worldwide. In the aftermath of the COVID-19 pandemic, these troubling statistics have worsened, with notable increases in suicidal behavior, especially among vulnerable populations such as the youth, the elderly, and those in socioeconomically disadvantaged groups. This paper aims to explore the impact of the COVID-19 pandemic on suicide rates in Spain, using a theoretical ex post facto analysis. Spain has witnessed an alarming rise in suicide rates, particularly among young people, and a disturbing trend of increased suicidal ideation and self-harm behaviors. While some studies report no significant change in suicide rates during the pandemic, others point to the exacerbating effects of social isolation, economic instability, and public health measures. This study provides an in-depth examination of the psychosocial consequences of the pandemic on mental health in Spain, emphasizing the urgency of the need to address pre-existing inequalities and implement effective suicide prevention measures. Furthermore, it highlights the importance of gender-sensitive strategies and the need for systemic reforms to ensure better mental healthcare access for all segments of society. To achieve this goal, this paper uses a narrative literature review combined with a theoretical ex post facto analysis to assess the influence of the COVID-19 pandemic on suicide patterns in Spain.
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Affiliation(s)
- Enrique Fernández-Vilas
- Department of Sociology and Social Work, University of Valladolid, José Tudela 12D, 42004 Soria, Spain
| | - Juan R Coca
- Department of Sociology and Social Work, University of Valladolid, José Tudela 12D, 42004 Soria, Spain
| | - Juan José Labora González
- Department of Political Science and Sociology, University of Santiago de Compostela, Ángel Jorge Echeverri, 15782 Santiago de Compostela, Spain
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de Araujo MC, Souza DDC, Dos Santos Rosa A, Dos Santos Ferreira VN, Mazzarino L, Veleirinho B, Miranda MD, Maraschin M. Green antiseptic for hand hygiene with high activity against SARS-CoV-2: Iota-carrageenan, quercetin, and Melaleuca alternifolia essential oil based nanoemulsion. Int J Biol Macromol 2025; 305:140920. [PMID: 39947550 DOI: 10.1016/j.ijbiomac.2025.140920] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2024] [Revised: 01/30/2025] [Accepted: 02/09/2025] [Indexed: 02/22/2025]
Abstract
The World Health Organization (WHO) has determined a series of guidelines to contain the advance and spread of COVID-19 and other influenza viruses. Among them, frequent hand hygiene has been widely recommended, resulting in an increased consumption of alcohol-based antiseptic products or synthetic molecules. However, when used in excess, these products might cause adverse consequences for human health, such as dermatitis, and for the environment, i.e., the selection of resistant bacterial genotypes. One of the alternatives to overcome this problem is the replacement of common antiseptics by formulations based on natural bioactive compounds with antimicrobial/antiviral activity. In addition, by nanostructuring formulations, it is possible to increase the bioavailability, stability, solubility, and absorption of bioactives in biological systems. In this sense, this study aimed to develop an antiseptic nanoemulsion based on natural bioactive compounds with virucidal activity against SARS-CoV-2. For that, oil-in-water (O/W) nanoemulsions were prepared, being the oil phase composed by Melaleuca alternifolia essential oil, quercetin, PEG400, and surfactants, while the aqueous phase presented carrageenan and purified water. Physicochemical characterization and stability studies were developed to evaluate the viability of the formulations over time. In addition, bactericidal activities against Staphylococcus aureus and antiviral activity against SARS-CoV-2 were determined by in vitro assays. As a result, the average size of the nanoparticles was recorded at 150 nm, with a Polydispersity Index (PdI) of 0.2 and a zeta potential around -10.0 mV. The stability of nanoformulations indicated the occurrence of quercetin-dependent creaming and sedimentation. In addition, the products presented a minimum shelf-life of 3 months. Regarding the bactericidal activity, a minimum inhibition concentration of 1.25 % for S. aureus was found. The cytotoxicity and antiviral assays revealed that the nano-based products showed 100 % of viral replication inhibition and proved to be safe for epithelial cells. In conclusion, two antiseptic nanoformulations with high anti-SARS-CoV-2 activity and great industrial and pharmacological potential were developed.
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Affiliation(s)
- Milena Conci de Araujo
- Plant Morphogenesis and Biochemistry Laboratory, Federal University of Santa Catarina, Florianópolis, Brazil
| | - Daniel Dias Coutinho Souza
- Laboratory of Morphology and Viral Morphogenesis, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil; Postgraduate Program in Cellular and Molecular Biology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | - Alice Dos Santos Rosa
- Laboratory of Morphology and Viral Morphogenesis, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil; Postgraduate Program in Cellular and Molecular Biology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil
| | | | - Letícia Mazzarino
- NanoScoping Solutions in Nanotechnology, Alpha Tech. Park, Florianópolis, Brazil
| | - Beatriz Veleirinho
- NanoScoping Solutions in Nanotechnology, Alpha Tech. Park, Florianópolis, Brazil
| | - Milene Dias Miranda
- Laboratory of Morphology and Viral Morphogenesis, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil; Postgraduate Program in Cellular and Molecular Biology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil.
| | - Marcelo Maraschin
- Plant Morphogenesis and Biochemistry Laboratory, Federal University of Santa Catarina, Florianópolis, Brazil.
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11
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Sui Q, Zhu C, Shi S, Xu J, Zhang J, Wang A, Chen P, Liang G, Zhang Y. Ganoderic acid A: an in-depth review of pharmacological effects and molecular docking analysis. JOURNAL OF ETHNOPHARMACOLOGY 2025; 349:119868. [PMID: 40316150 DOI: 10.1016/j.jep.2025.119868] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/20/2024] [Revised: 03/24/2025] [Accepted: 04/22/2025] [Indexed: 05/04/2025]
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE Ganoderic acid A (GAA, C30H44O7) is one of the most abundant and active components of Ganoderic acids (GAs). GAs are highly oxidized tetracyclic triterpenoid compounds mainly derived from Ganoderma lucidum (Curtis) P. Karst (Chinese: ). GAA is primarily isolated from the fruiting body of Ganoderma lucidum. Modern pharmacological investigations have established the broad pharmacological effects of GAA, highlighting its notable influence on managing various conditions, including inflammatory diseases, neurodegenerative diseases, and cancer. This review provides a comprehensive summary of GAA's pharmacological activities. MATERIAL AND METHODS The literature in this review were searched in PubMed and China National Knowledge Infrastructure (CNKI) using the keywords "Ganoderic acid A″, "Pharmacology" and "Pharmacokinetics". The literature cited in this review dates from 2000 to 2024. RESULTS According to the data, GAA exerts anti-inflammatory, antioxidant, antitumor, neuropsychopharmacological, hepatoprotective, cardiovascular, renoprotective, and lung protective effects by regulating a variety of signal transduction pathways, such as nuclear factor kappa-B (NF-κB), Janus kinase/signal transducer and activator of transcription (JAK/STAT), Toll-like receptor 4 (TLR4), nuclear factor erythroid 2-related factor-2 (Nrf2), phosphoinositide-3-kinase (PI3K)/AKT, mammalian target of rapamycin (mTOR), mitogen-activated protein kinase (MAPK), and Notch. Given its promising pharmacological activity, GAA holds excellent potential for treating human diseases. The pharmacokinetic properties of GAA have also been reviewed, revealing low bioavailability but high absorption and elimination rates. In addition, network pharmacology and molecular docking analyses verified that GAA plays a role in multiple diseases through MAPK3, tumor necrosis factor (TNF), caspase-3 (CASP3), peroxisome proliferator-activated receptor gamma (PPARG), and β-catenin (CTNNB1) signaling pathways. CONCLUSION GAA plays a pivotal role in various pathological and physiological processes, boasting broad application prospects. Furthermore, the network pharmacological results reveal the mechanisms of GAA in the treatment of multiple diseases. In the future, it is necessary to conduct further experiments to elucidate its specific mechanism of action, thus laying the foundation for the scientific utilization of GAA.
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Affiliation(s)
- Qi Sui
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China
| | - Chengkai Zhu
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China
| | - Sha Shi
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China
| | - Jiaqi Xu
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China
| | - Jingnan Zhang
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China
| | - Ao Wang
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Department of Pharmacy, School of Medicine, Hangzhou City University, 50 Huzhou Rd, Hangzhou, Zhejiang, 310015, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China
| | - Peng Chen
- Department of Pharmacy, School of Medicine, Hangzhou City University, 50 Huzhou Rd, Hangzhou, Zhejiang, 310015, China.
| | - Guang Liang
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, 325035, China.
| | - Yi Zhang
- School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China; Zhejiang TCM Key Laboratory of Pharmacology and Translational Research of Natural Products, Hangzhou Medical College, Hangzhou, Zhejiang, 310012, China.
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12
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Concha-Toloza M, Collado González L, Herrera Estrella A, Fernández Do Porto D, Campos Sánchez R, Molina-Mora JA. Genomic, socio-environmental, and sequencing capability patterns in the surveillance of SARS-CoV-2 in Latin America and the Caribbean up to 2023. Sci Rep 2025; 15:14607. [PMID: 40287493 PMCID: PMC12033298 DOI: 10.1038/s41598-025-98829-9] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2024] [Accepted: 04/15/2025] [Indexed: 04/29/2025] Open
Abstract
Latin America and the Caribbean (LAC) have been one of the most strongly impacted regions by the Coronavirus disease 2019 (COVID-19) emergency, with more than 83 million reported infections and 1.7 million deaths until December 2023, intensifying economic and social inequalities. This study aimed to provide information to the regional SARS-CoV-2 surveillance programs by determining genomic, socio-environmental, and sequencing capacity patterns associated with the circulation of the virus in Latin America up to 2023. Data from 24 countries in LAC were analyzed using public databases up to December 2023. A sampling of SARS-CoV-2 sequences from cases in the region enabled a phylogenomic study to elucidate the temporal distribution of various variants of concern (VOCs), mutations, and recombinants. Also, we identified differences in sequencing capabilities in LAC. Additionally, correlation and generalized linear model (GLM) analyses were conducted to explore potential associations between 89 socio-environmental variables and five COVID-19 indicators at the country level. The phylogenomic analyses revealed a diversity of variants with the predominance of some during specific periods, mainly VOCs and some recombinant cases, and a mutation rate of 8.39 × 10-4 substitutions per site per year, which are in line with other regions of the world. Besides, a low sequencing rate in LAC (on average 0.7% of cases) and incomplete databases in several countries were identified. In the analysis of indicators, correlations between 9 socio-environmental indicators and four COVID-19 variables associated with cases, deaths, and diagnostic tests related to the virus in the region, although not for sequencing percentages. This study provides information about the development of COVID-19 disease in LAC in terms of the viral genome, sequencing capabilities, and the region's complex socio-environmental conditions. Therefore, emphasis must be placed on implementing an integrated epidemiological surveillance approach to strengthen public health infrastructure and improve cooperation and preparedness for future infections affecting this region.
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Affiliation(s)
- Macarena Concha-Toloza
- Instituto de Bioquímica y Microbiología, Facultad de Ciencias, Universidad Austral de Chile, 5110566, Valdivia, Chile
| | - Luis Collado González
- Instituto de Bioquímica y Microbiología, Facultad de Ciencias, Universidad Austral de Chile, 5110566, Valdivia, Chile
| | | | - Darío Fernández Do Porto
- Instituto de Cálculo, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires CONICET, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
| | - Rebeca Campos Sánchez
- Centro de Investigación en Biología Celular y Molecular (CIBCM), Universidad de Costa Rica, San José, Costa Rica
| | - Jose Arturo Molina-Mora
- Facultad de Microbiología, Centro de Investigación en Enfermedades Tropicales (CIET) and Centro de investigación en Hematología y Trastornos Afines (CIHATA), Universidad de Costa Rica, San José, Costa Rica.
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13
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Bamrung P, Toviwek B, Samsudin F, Chairatana P, Bond PJ, Pongprayoon P. The Binding of Brazilin from C. sappan to the Full-Length SARS-CoV-2 Spike Proteins. Int J Mol Sci 2025; 26:4100. [PMID: 40362339 PMCID: PMC12072004 DOI: 10.3390/ijms26094100] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2025] [Revised: 04/15/2025] [Accepted: 04/23/2025] [Indexed: 05/15/2025] Open
Abstract
The emergence of coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become a global issue since 2019. The prominent characteristic of SARS-CoV-2 is the presence of the spike (S) protein protruding from the virus particle envelope. The S protein is a major drug and vaccine target because it initiates the key step in infection. Medicinal herbs are a potential treatment option to enhance immunity to fight viral infections. Caesalpinia sappan L. has been reported to display promising anti-viral activities. Specifically, brazilin (BRA), a major bioactive compound in C. sappan, was reported to play a role in inhibiting viral infection. Thus, the ability of BRA as a COVID-19 treatment was tested. The S protein was used as the BRA target of this work. Understanding the binding mechanism of BRA to the S protein is crucial for future utilisation of C. sappan as a COVID-19 treatment or other coronavirus-caused pandemics. Here, we performed molecular docking of BRA onto the S protein receptor binding domain (RBD) and multimerisation (MM) pockets. Molecular dynamics (MD) simulations were conducted to study the stability of binding to glycosylated and non-glycosylated S protein constructs. BRA can bind to the Receptor-binding motif (RBM) on an RBD surface stably; however, it is too large to fit into the MM pocket, resulting in dissociation. Nonetheless, BRA is bound by residues near the S1/S2 interface. We found that glycosylation has no effect on BRA binding, as the proposed binding site is far from any glycans. Our results thus indicate that C. sappan may act as a promising preventive and therapeutic alternative for COVID-19 treatment.
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Affiliation(s)
- Phonphiphat Bamrung
- Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand; (P.B.); (B.T.)
| | - Borvornwat Toviwek
- Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand; (P.B.); (B.T.)
| | - Firdaus Samsudin
- Bioinformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), 30 Biopolis Street, #07-01 Matrix, Singapore 138671, Singapore;
| | - Phoom Chairatana
- Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand;
| | - Peter John Bond
- Bioinformatics Institute (BII), Agency for Science, Technology and Research (A*STAR), 30 Biopolis Street, #07-01 Matrix, Singapore 138671, Singapore;
- Department of Biological Sciences, National University of Singapore, Singapore 117543, Singapore
| | - Prapasiri Pongprayoon
- Department of Chemistry, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand; (P.B.); (B.T.)
- Center for Advanced Studies in Nanotechnology for Chemical, Food and Agricultural Industries, KU Institute for Advanced Studies, Kasetsart University, Bangkok 10900, Thailand
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14
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Erdmann M, Wing PAC, Webb I, Kavanagh Williamson M, Jearanaiwitayakul T, Sullivan E, Bazire J, Shytaj IL, McKeating JA, Matthews DA, Davidson AD. A Novel Toolkit of SARS-CoV-2 Sub-Genomic Replicons for Efficient Antiviral Screening. Viruses 2025; 17:597. [PMID: 40431609 DOI: 10.3390/v17050597] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/11/2025] [Revised: 04/02/2025] [Accepted: 04/14/2025] [Indexed: 05/29/2025] Open
Abstract
SARS-CoV-2 is classified as a containment level 3 (CL3) pathogen, limiting research access and antiviral testing. To address this, we developed a non-infectious viral surrogate system using reverse genetics to generate sub-genomic replicons. These replicons contained the nsp1 mutations K164A and H165A and had the spike, membrane, ORF6, and ORF7a coding sequences replaced with various reporter and selectable marker genes. Replicons based on the ancestral Wuhan Hu-1 strain and the Delta variant of concern were replication-competent in multiple cell lines, as assessed by Renilla luciferase activity, fluorescence, immunofluorescence staining, and single-molecule fluorescent in situ hybridization. Antiviral assays using transient replicon expression showed that remdesivir effectively inhibited both replicon and viral replication. Ritonavir and cobicistat inhibited Delta variant replicons similarly to wild-type virus but did not inhibit Wuhan Hu-1 replicon replication. To further investigate the impact of nsp1 mutations, we generated a recombinant SARS-CoV-2 virus carrying the K164A and H165A mutations. The virus exhibited attenuated replication across a range of mammalian cell lines, was restricted by the type I interferon response, and showed reduced cytopathic effects. These findings highlight the utility of sub-genomic replicons as reliable CL2-compatible surrogates for studying SARS-CoV-2 replication and drug activity mechanisms.
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Affiliation(s)
- Maximilian Erdmann
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | - Peter A C Wing
- Chinese Academy of Medical Sciences Oxford Institute, University of Oxford, Oxford OX3 7BN, UK
- Nuffield Department of Medicine, University of Oxford, Oxford OX3 7BN, UK
| | - Isobel Webb
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | | | - Tuksin Jearanaiwitayakul
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
- Faculty of Science, Mahidol University, Bangkok 10400, Thailand
| | - Edward Sullivan
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | - James Bazire
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | - Iart Luca Shytaj
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | - Jane A McKeating
- Chinese Academy of Medical Sciences Oxford Institute, University of Oxford, Oxford OX3 7BN, UK
- Nuffield Department of Medicine, University of Oxford, Oxford OX3 7BN, UK
| | - David A Matthews
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
| | - Andrew D Davidson
- School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK
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15
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Kim CW, Ku KB, Hwang I, Jung HE, Kim KD, Lee HK. Differential responses of lung and intestinal microbiota to SARS-CoV-2 infection: a comparative study of the Wuhan and Omicron strains in K18-hACE2 Tg mice. Lab Anim Res 2025; 41:11. [PMID: 40270072 PMCID: PMC12016229 DOI: 10.1186/s42826-025-00241-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2024] [Revised: 03/07/2025] [Accepted: 04/06/2025] [Indexed: 04/25/2025] Open
Abstract
BACKGROUND The COVID-19 pandemic, caused by SARS-CoV-2, has led to the emergence of viral variants with distinct characteristics. Understanding the differential impacts of SARS-CoV-2 variants is crucial for effective public health response and treatment development. We investigated the differential effects of the original Wuhan strain and the emergent Omicron variant of SARS-CoV-2 using a K18-hACE2 transgenic mouse model. We compared the mortality rates, viral loads, and histopathological changes in lung and tracheal tissues, as well as alterations in the lung and intestinal microbiota following infection. RESULTS Our findings revealed significant differences between the variants, with the Wuhan strain causing higher mortality rates, severe lung pathology, and elevated viral loads compared to the Omicron variant. Microbiome analyses uncovered novel and distinct shifts in the lung and intestinal microbiota associated with each variant, providing evidence for variant-specific microbiome alterations. These changes suggest microbiome-related mechanisms that might modulate disease severity and host responses to SARS-CoV-2 infection. CONCLUSIONS This study highlights critical differences between the Wuhan strain and Omicron variant in terms of mortality, lung pathology, and microbiota changes, emphasizing the role of the microbiome in influencing disease outcomes. Novel findings include the identification of variant-specific microbiota shifts, which underscore potential microbiome-related mechanisms underlying differences in disease severity. These insights pave the way for future research exploring microbiome-targeted interventions to mitigate the impacts of SARS-CoV-2 and other viral infections.
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Affiliation(s)
- Chae Won Kim
- Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Republic of Korea
- Life Science Institute, KAIST, Daejeon, 34141, Republic of Korea
| | - Keun Bon Ku
- Center for Infectious Disease Vaccine and Diagnosis Innovation, Korea Research Institute of Chemical Technology, Daejeon, 34114, Republic of Korea
| | - Insu Hwang
- Center for Infectious Disease Vaccine and Diagnosis Innovation, Korea Research Institute of Chemical Technology, Daejeon, 34114, Republic of Korea
| | - Hi Eun Jung
- Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Republic of Korea
- Life Science Institute, KAIST, Daejeon, 34141, Republic of Korea
| | - Kyun-Do Kim
- Center for Infectious Disease Vaccine and Diagnosis Innovation, Korea Research Institute of Chemical Technology, Daejeon, 34114, Republic of Korea.
| | - Heung Kyu Lee
- Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Republic of Korea.
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16
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Groves D, Cunnison R, McMahon A, Fan H, Sharps J, Deng A, Keown JR, Fodor E, Robb NC. Mechanistic insights into the activity of SARS-CoV-2 RNA polymerase inhibitors using single-molecule FRET. Nucleic Acids Res 2025; 53:gkaf351. [PMID: 40298114 PMCID: PMC12038395 DOI: 10.1093/nar/gkaf351] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2024] [Revised: 04/08/2025] [Accepted: 04/16/2025] [Indexed: 04/30/2025] Open
Abstract
The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has resulted in significant global mortality, with over 7 million cases reported. Despite extensive research and high vaccination rates, highly mutated forms of the virus continue to circulate. It is therefore important to understand the viral lifecycle and the precise molecular mechanisms underlying SARS-CoV-2 replication. To address this, we developed a single-molecule Förster resonance energy transfer (smFRET) assay to directly visualize and analyse in vitro RNA synthesis by the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp). We purified the minimal replication complex, comprising nsp12, nsp7, and nsp8, and combined it with fluorescently labelled RNA substrates, enabling real-time monitoring of RNA primer elongation at the single-molecule level. This platform allowed us to investigate the mechanisms of action of key inhibitors of SARS-CoV-2 replication. In particular, our data provides evidence for remdesivir's mechanism of action, which involves polymerase stalling and subsequent chain termination dependent on the concentration of competing nucleotide triphosphates. Our study demonstrates the power of smFRET to provide dynamic insights into SARS-CoV-2 replication, offering a valuable tool for antiviral screening and mechanistic studies of viral RdRp activity.
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Affiliation(s)
- Danielle Groves
- Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom
| | - Rory Cunnison
- Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom
| | - Andrew McMahon
- Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom
| | - Haitian Fan
- Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3PU, United Kingdom
| | - Jane Sharps
- Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3PU, United Kingdom
| | - Adrian Deng
- School of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom
| | - Jeremy R Keown
- School of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom
| | - Ervin Fodor
- Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3PU, United Kingdom
| | - Nicole C Robb
- Warwick Medical School, University of Warwick, Coventry, CV4 7AL, United Kingdom
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17
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Sun R, Xia L, She G, Li J, Wang Y, Chen Y, Yang Q, Zhang S, Liu F, Chen Y, Zhang L, Zhang C, Lv W, Huang E, Zhang L. Repeated-dose toxicity and immunogenicity evaluation of a recombinant subunit COVID-19 vaccine (ZF2001) in rats. Front Cell Infect Microbiol 2025; 15:1548787. [PMID: 40330020 PMCID: PMC12053236 DOI: 10.3389/fcimb.2025.1548787] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/20/2024] [Accepted: 03/24/2025] [Indexed: 05/08/2025] Open
Abstract
Coronavirus disease 19 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), had given rise to a massive epidemic. Owing to the high morbidity and mortality of COVID-19 and the lack of effective therapies, safe and effective vaccination is the optimum choice for controlling this epidemic and preventing infection. The protein subunit vaccine ZF2001, which targets the receptor-binding domain (RBD) protein of SARS-CoV-2, has a significant protective effect against COVID-19. At the beginning of the COVID-19 epidemic, to promote the early approval of ZF2001 for clinical trials by the National Medical Products Administration of China (NMPA), a comprehensive evaluation of its toxicity in vivo was warranted. In the present study, a major part of the above series of studies, we evaluated the safety, immunogenicity and efficacy of the ZF2001 vaccine for the first time in adult Sprague Dawley (SD) rats. The male and female rats were administered three doses of the ZF2001 vaccine (25 μg or 50 μg NCP-RBD protein/dose, containing the aluminum-based adjuvant). The safety profile of ZF2001 was assessed by observing the general health status, local toxicity at the site of administration, immunotoxicity, immunogenicity, blood chemistry and hematology parameters in SD rats. In general, our results indicated that the ZF2001 vaccine did not induce significant systemic toxicity in rats, with a no-observed adverse effect level (NOAEL) of 50 μg NCP-RBD protein/rat. Moreover, the ZF2001 vaccine showed good immunogenicity by inducing the production of specific IgG antibodies in rats after three consecutive immunizations. In addition, histological examination revealed recoverable inflammatory changes in quadricep muscles and adjacent lymph nodes at the vaccine injection site. In summary, our systematic toxicology study proves the safety, tolerability and immunogenicity of the ZF2001 vaccine, which further supports the results of clinical trials of ZF2001.
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MESH Headings
- Animals
- COVID-19 Vaccines/immunology
- COVID-19 Vaccines/administration & dosage
- COVID-19 Vaccines/toxicity
- COVID-19 Vaccines/adverse effects
- Male
- Rats
- Female
- Rats, Sprague-Dawley
- SARS-CoV-2/immunology
- COVID-19/prevention & control
- COVID-19/immunology
- Vaccines, Subunit/immunology
- Vaccines, Subunit/administration & dosage
- Vaccines, Subunit/toxicity
- Vaccines, Subunit/adverse effects
- Antibodies, Viral/blood
- Vaccines, Synthetic/immunology
- Vaccines, Synthetic/administration & dosage
- Immunogenicity, Vaccine
- Antibodies, Neutralizing/blood
- Spike Glycoprotein, Coronavirus/immunology
- Immunoglobulin G/blood
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Affiliation(s)
- Ruimin Sun
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China
| | - Lijuan Xia
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China
| | - Guangbiao She
- Anhui Zhifei Longcom Biopharmaceutical Co., Ltd., HeFei, China
- Recombinant Vaccine Research and Development Joint Laboratory of Anhui Province, HeFei, China
| | - Jinrong Li
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Yiru Wang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Faculty of Chinese Medicine, Macau University of Science and Technology, Macao, Macao SAR, China
| | - Yunxiang Chen
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China
| | - Qian Yang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Siming Zhang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Fang Liu
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Ying Chen
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Liyan Zhang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Chengda Zhang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
| | - Wanqiang Lv
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China
| | - Enqi Huang
- Anhui Zhifei Longcom Biopharmaceutical Co., Ltd., HeFei, China
- Recombinant Vaccine Research and Development Joint Laboratory of Anhui Province, HeFei, China
| | - Lijiang Zhang
- Key Laboratory of Drug Safety Evaluation and Research of Zhejiang Province, Center of Safety Evaluation and Research, Hangzhou Medical College, Hangzhou, China
- Engineering Research Center of Novel Vaccine of Zhejiang Province, Hangzhou Medical College, Hangzhou, China
- Qingshan Lake Science and Technology Innovation Center, Hangzhou Medical College, Hangzhou, China
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Bertola B, Cotolí-Crespo A, San Onofre N, Soriano JM. The Mystery of Certain Lactobacillus acidophilus Strains in the Treatment of Gastrointestinal Symptoms of COVID-19: A Review. Microorganisms 2025; 13:944. [PMID: 40284780 PMCID: PMC12029412 DOI: 10.3390/microorganisms13040944] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/18/2025] [Revised: 04/08/2025] [Accepted: 04/17/2025] [Indexed: 04/29/2025] Open
Abstract
COVID-19 presents a wide range of symptoms, including gastrointestinal manifestations such as diarrhea, nausea, and abdominal pain. Lactobacillus acidophilus has been proposed as a potential adjunct therapy to alleviate these symptoms due to its probiotic properties, which help restore gut microbiota balance and modulate immune responses. This review systematically analyzed studies assessing the effects of L. acidophilus in COVID-19 patients with gastrointestinal symptoms. The literature search was conducted through PubMed and the WHO COVID-19 database using keywords such as "Lactobacillus acidophilus", "COVID-19", "gastrointestinal symptoms", and "inflammation markers". The search covered studies published until February 2025. Inclusion criteria: observational and clinical trials with L. acidophilus for symptom relief. Exclusion: animal studies and non-ethical approvals. The findings suggest that L. acidophilus supplementation may contribute to faster resolution of diarrhea, improved gut microbiota balance, and reduced inflammatory markers. However, some studies have found no significant impact on hospitalization rates or disease progression. The probiotic's mechanisms of action appear to involve microbiota modulation, intestinal barrier reinforcement, and anti-inflammatory effects rather than direct viral inhibition in COVID-19 after progression. Some L. acidophilus strains show promise, and clinical validation should follow careful preclinical studies (in vitro, cell lines, and animal models), especially in vulnerable populations such as immunocompromised individuals. Understanding the gut-lung axis and its role in immune response regulation, together with the need for a thorough characterization of the specific strains, including biochemical, genomic, and functional properties, before testing in humans, may provide deeper insights into the therapeutic potential of probiotics in viral infections.
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Affiliation(s)
- Belén Bertola
- Food & Health Lab, Institute of Materials Science, University of Valencia, 46980 Paterna, Spain;
| | - Amparo Cotolí-Crespo
- Faculty of Psychology and Speech Therapy, University of Valencia, 46010 Valencia, Spain;
| | - Nadia San Onofre
- NUTRALiSS Research Group, Faculty of Health Sciences, Universitat Oberta de Catalunya, Rambla del Poblenou 156, 08018 Barcelona, Spain;
| | - Jose M. Soriano
- Food & Health Lab, Institute of Materials Science, University of Valencia, 46980 Paterna, Spain;
- Joint Research Unit on Endocrinology, Nutrition and Clinical Dietetics, University of Valencia-Health Research Institute La Fe, 46026 Valencia, Spain
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19
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dos Reis GG, Silvestre RT, Alves G, Delmonico L, Chantre-Justino M, Moreira ADS, Müller BDLA, do Nascimento CR, da Silva DLP, dos Santos LS, de Mattos-Guaraldi AL, Ornellas MH. Leukocyte telomere length and telomerase activity in Long COVID patients from Rio de Janeiro, Brazil. Mem Inst Oswaldo Cruz 2025; 120:e240129. [PMID: 40243865 PMCID: PMC11984962 DOI: 10.1590/0074-02760240129] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/20/2024] [Accepted: 12/03/2024] [Indexed: 04/18/2025] Open
Abstract
BACKGROUND Coronavirus disease 2019 (COVID-19) is caused by the new coronavirus 2 (severe acute respiratory syndrome coronavirus 2 - SARS-CoV-2). Long COVID is a new condition associated with persistent COVID-19 symptoms and/or new emerging symptoms. Telomeres are specialised structures for genome protection at the end of chromosomes and telomerase is the enzyme that synthesises telomere DNA. OBJECTIVES Patients with Long COVID symptoms were recruited at the Pedro Ernesto University Hospital (HUPE) in Rio de Janeiro, Brazil, with the main purpose of investigating the association between telomere length and Long COVID. METHODS Leukocyte telomere length (LTL) was determined by quantitative real-time polymerase chain reaction (qPCR) in 34 Long COVID patients compared to a control group (n = 122). Telomerase activity was determined by qPCR assays using the commercial kit from ScienCell. A questionnaire on symptoms, vaccine doses and blood count was completed. FINDINGS The Long COVID patients were found to have an increase in LTL. Telomerase activity was also examined in a smaller number of patients and found to be reactivated in the blood. MAIN CONCLUSIONS It will be necessary to conduct further studies and monitor Long COVID patients to determine if future health issues could be linked to telomerase activity and elongated telomeres.
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Affiliation(s)
- Giane Gonçalves dos Reis
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Pós-Graduação em Ciências Médicas, Rio de Janeiro, RJ, Brasil
| | - Rafaele Tavares Silvestre
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Pós-Graduação em Ciências Médicas, Rio de Janeiro, RJ, Brasil
| | - Gilda Alves
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Departamento de Microbiologia, Imunologia e Parasitologia, Rio de Janeiro, RJ, Brasil
| | - Lucas Delmonico
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
| | - Mariana Chantre-Justino
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
- Instituto Nacional de Traumatologia e Ortopedia, Rio de Janeiro, RJ, Brasil
| | - Aline dos Santos Moreira
- Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Genômica Aplicada e Bioinovações, Rio de Janeiro, RJ, Brasil
- Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Rede de Plataformas Tecnológicas Fiocruz, Plataforma de Sequenciamento de Nova Geração, Rio de Janeiro, RJ, Brasil
| | - Beatriz de Lima Alessio Müller
- Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Genômica Aplicada e Bioinovações, Rio de Janeiro, RJ, Brasil
- Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Rede de Plataformas Tecnológicas Fiocruz, Plataforma de Sequenciamento de Nova Geração, Rio de Janeiro, RJ, Brasil
| | - Carolina Ramos do Nascimento
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
| | - Denzel Luis Pereira da Silva
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
| | - Louisy Sanches dos Santos
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Departamento de Microbiologia, Imunologia e Parasitologia, Rio de Janeiro, RJ, Brasil
| | - Ana Luíza de Mattos-Guaraldi
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Departamento de Microbiologia, Imunologia e Parasitologia, Rio de Janeiro, RJ, Brasil
| | - Maria Helena Ornellas
- Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas, Laboratório de Marcadores Circulantes, Rio de Janeiro, RJ, Brasil
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Zhao JY, Chu KK, Han PY, Yang Z, Tang Y, Kong W, Long Y, Zong LD, Ge XY, Zhang YZ. Detection of Coronaviruses and Genomic Characterization of Gammacoronaviruses from Overwintering Black-Headed Gulls ( Chroicocephalus ridibundus) in Yunnan Province, China. Microorganisms 2025; 13:874. [PMID: 40284710 PMCID: PMC12029519 DOI: 10.3390/microorganisms13040874] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2025] [Revised: 04/01/2025] [Accepted: 04/05/2025] [Indexed: 04/29/2025] Open
Abstract
Black-headed gulls have been confirmed as the natural hosts of Deltacoronavirus (δ-CoV) and Gammacoronavirus (γ-CoV). A total of 59 CoV-PCR-positive fecal samples were identified among 509 fecal samples collected from overwintering black-headed gulls in Yunnan Province, China. The prevalence of black-headed gull deltacoronavirus (BHG-DCoV) was 3.54% (18/509), while that of black-headed gull gammacoronavirus (BHG-GCoV) was 8.06% (41/509). The prevalence of BHG-GCoV was significantly higher than that of BHG-DCoV (χ2 = 9.518, p < 0.01). Two complete genome sequences of BHG-GCoVs were obtained, with lengths of 27,358 bp and 27,355 bp, respectively, from the fecal samples of black-headed gulls. The nucleotide similarity between the two complete genomes is 98.75%. Phylogenetic analysis based on the whole genome has confirmed that the two strains of BHG-GCoVs clustered into the species Gammacoronavirus anatis. Although BHG-GCoVs belong to the species Gammacoronavirus anatis, they are distantly related to the representative strain Duck_CoV 2714 and exhibit a closer genetic relationship with GCoVs from Xenus cinereus (AvXc-GCoV) and Numenius phaeopus (AvNp-GCoV). Similarity analysis of the five conserved domains revealed a high amino acid similarity not only with AvXc-GCoV and AvNp-GCoV but also with GCoVs from common gulls detected in Poland and those from ruddy turnstones identified in Australia. Additionally, we found that, except for the common gull, the amino acid sequences of the S protein of BHG-GCoVs showed a 88.69% to 96.44% similarity with those of GCoVs carried by Charadriiformes, while the similarity with GCoVs carried by Anseriformes ranged from 31.15% to 54.81%. Furthermore, recombination events were detected in BHG-GCoVs, suggesting that these strains are likely recombinant strains of common gull GCoV and the GCoV of Arenaria interpres (AvAi-GCoV), indicating that recombination events may occur frequently among GCoVs.
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Affiliation(s)
- Jun-Ying Zhao
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Kan-Kan Chu
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Pei-Yu Han
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Ze Yang
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Yi Tang
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Wei Kong
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Yun Long
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Li-Dong Zong
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
| | - Xing-Yi Ge
- Hunan Provincial Key Laboratory of Medical Virology, Institute of Pathogen Biology and Immunology, College of Biology, Hunan University, Changsha 410012, China
| | - Yun-Zhi Zhang
- Yunnan Key Laboratory of Screening and Research on Anti-Pathogenic Plant Resources from Western Yunnan, Key Laboratory for Cross-Border Control and Quarantine of Zoonoses in Universities of Yunnan Province, Institute of Preventive Medicine, School of Public Health, Dali University, Dali 671000, China; (J.-Y.Z.); (K.-K.C.); (P.-Y.H.); (Z.Y.); (Y.T.); (W.K.); (Y.L.); (L.-D.Z.)
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21
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Liu Y, Li C, Wu Z, Zhao Y, Yin T, Liu X, Hui J, Wang Q, Pan Y, Shan Y, Qu X. Self-assembled epitope-based nanoparticles targeting the SARS-CoV-2 spike protein enhanced the immune response and induced potential broad neutralizing activity. Front Cell Infect Microbiol 2025; 15:1560330. [PMID: 40270771 PMCID: PMC12014594 DOI: 10.3389/fcimb.2025.1560330] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/14/2025] [Accepted: 03/20/2025] [Indexed: 04/25/2025] Open
Abstract
Introduction The ongoing COVID-19 has caused a global pandemic, resulting in millions of infections and deaths. While current vaccines target the SARS-CoV-2 spike (S) protein, its high mutation rate significantly compromises vaccine efficacy. We aimed to evaluate the potential of epitope-based nanoparticles (NPs) to induce broad cross-protection and durable immune responses against SARS-CoV-2. Methods Four conserved epitopes derived from the receptor-binding domain (RBD) and S2 subunit of the spike protein were integrated into Helicobacter pylori ferritin to create epitope-based NPs named S18-F, RBM-F, UH-F, and HR2-F. The immunogenicity of the epitope-based NPs was evaluated through animal experiments to measure epitope-specific antibody titers and assess neutralizing activity against SARS-CoV-2 pseudovirus. To characterize cellular immune responses, splenic lymphocyte proliferation following epitope stimulation was measured, and cytokine secretion profiles including IFN-γ, IL-2, IL-4, and IL-10 were analyzed to determine Th1/Th2 immune polarization. Antibody-dependent cellular cytotoxicity (ADCC) assays were performed to evaluate NP-enhanced recognition and elimination of infected target cells. Results These NPs induced high titers of epitope-specific antibodies lasting three months post-immunization. Sera from the RBM-F, UH-F, and HR2-F groups exhibited neutralizing activity against the SARS-CoV-2 pseudovirus WH-1 in vitro. Splenic lymphocytes from the S18-F, RBM-F, and UH-F groups showed significantly increased proliferation. Lymphocytes from the RBM-F group demonstrated increased secretion of IFN-γ, IL-2, IL-4, and IL-10 cytokines, indicating a balanced Th1 and Th2 immune response. Immune sera from the S18-F and mixed-immunized groups exhibited antibody-dependent cellular cytotoxicity. Discussion The results indicate that these NPs induce robust humoral and cellular immune responses, potentially offering a promising strategy for effective vaccine development against SARS-CoV-2.
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Affiliation(s)
- Yue Liu
- Department of Echocardiography, The First Hospital of Jilin University, Changchun, China
- Institute of Virology and AIDS Research, The First Hospital of Jilin University, Changchun, China
| | - Chenxi Li
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Zirui Wu
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Yu Zhao
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Tieyan Yin
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Xiaopan Liu
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Jiaru Hui
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Qingyu Wang
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Yi Pan
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
| | - Yaming Shan
- National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, China
- Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun, China
| | - Xinglong Qu
- Department of Respiratory, The First Hospital of Jilin University, Changchun, China
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22
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Ferrarezi AA, de Souza JVP, Maigret B, Kioshima ÉS, Moura S, de Oliveira AJB, Rosa FA, Gonçalves RAC. Rational design and synthesis of pyrazole derivatives as potential SARS-CoV-2 M pro inhibitors: An integrated approach merging combinatorial chemistry, molecular docking, and deep learning. Bioorg Med Chem 2025; 120:118095. [PMID: 39929031 DOI: 10.1016/j.bmc.2025.118095] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/03/2024] [Revised: 01/17/2025] [Accepted: 02/02/2025] [Indexed: 02/12/2025]
Abstract
The global impact of SARS-CoV-2 has highlighted the urgent need for novel antiviral therapies. This study integrates combinatorial chemistry, molecular docking, and deep learning to design, evaluate and synthesize new pyrazole derivatives as potential inhibitors of the SARS-CoV-2 main protease (Mpro). A library of over 60,000 pyrazole-based structures was generated through scaffold decoration to enhance chemical diversity. Virtual screening employed molecular docking (ChemPLP scoring) and deep learning (DeepPurpose), with consensus ranking to identify top candidates. Binding free energy calculations refined the selection, revealing critical structural features such as tryptamine and N-phenyl fragments for Mpro binding. High-temperature solvent-free amidation allowed the synthesis of a selected derivative. Final compounds demonstrated favorable drug-likeness properties based on Lipinski's and Veber's rules. This work highlights the integration of computational and synthetic strategies to accelerate the discovery of Mpro inhibitors and provides a framework for future antiviral development.
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Affiliation(s)
- Arthur Antunes Ferrarezi
- Programa de Pós-graduação em Ciências Farmacêuticas, Departamento de Farmácia, Universidade Estadual de Maringá, Maringá, PR, Brazil
| | - João Vítor Perez de Souza
- Division of Translational Health Sciences, Department of Emergency Medicine, Duke University School of Medicine, United States of America
| | - Bernard Maigret
- Université de Lorraine, Centre Nationale de la Recherche Scientifique, Nancy, France
| | - Érika Seki Kioshima
- Programa de Pós-Graduação em Biociências e Fisiopatologia, Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá, Maringá, PR, Brazil
| | - Sidnei Moura
- Instituto de Biotecnologia, Universidade de Caxias do Sul (UCS), 95070-560 Caxias do Sul, RS, Brazil
| | - Arildo José Braz de Oliveira
- Programa de Pós-graduação em Ciências Farmacêuticas, Departamento de Farmácia, Universidade Estadual de Maringá, Maringá, PR, Brazil
| | - Fernanda Andreia Rosa
- Programa de Pós-Graduação em Química, Departamento de Química, Universidade Estadual de Maringá, Maringá, PR, Brazil.
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23
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Çelik G, Karaoğlu ŞA, Suyabatmaz Ş, Bozdeveci A, Yılmaz GT, Yaylı N, Akpınar R, Çiçek AÇ. Synthesis, biological evaluation and molecular docking studies of flavonol-3-O-β-D-glycoside as a potential inhibitor of SARS-CoV-2 main protease (3CLpro) in drug development for COVID-19. Int J Biol Macromol 2025; 298:139621. [PMID: 39818399 DOI: 10.1016/j.ijbiomac.2025.139621] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/07/2024] [Revised: 01/06/2025] [Accepted: 01/06/2025] [Indexed: 01/18/2025]
Abstract
The COVID-19 pandemic began in March 2020 and has affected many countries and infected over a million people. It has had a serious impact on people's physical and mental health, daily life and the global economy. Today, many drugs show limited efficacy in the treatment of COVID-19 and studies to develop effective drugs continue. Here, we aim to the synthesise and characterise of the flavonol-3-O-glycoside derivatives, the following and evaluated molecular docking studies with antimicrobial activity, inhibition of SARS-CoV-2 main protease enzyme (3CLpro) and nuclease activity. Molecular docking simulations of the synthesized flavonol-3-O-glycoside derivatives, especially compounds 5a, 5d, 5h, 5i and 5m, showed a stronger interaction with SARS-CoV-2 3CLpro in the active site. Two compounds from the target compounds, 5h and 5m, were found to be specifically effective against M. smegmatis and yeasts. In particular, compounds 5a, 5d, 5h, 5i and 5m, which exhibited high activity against the SARS-CoV-2 main protease enzyme, were found to be effective at low concentrations. We determined the IC50 values for the compounds that showed an inhibitory effect as well as their nuclease activities, which further emphasising the potential of our results. Among these, compound 5d showed a significant competitive inhibitor of 3CLpro. Furthermore, nuclease activity studies identified compound 5d as the most potent. The above results suggest that the flavonol-3-O-glycoside derivatives could be promising new antiviral agents for the development of 3CLpro inhibitors to combat COVID-19.
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Affiliation(s)
- Gonca Çelik
- Department of Chemistry, Faculty of Science, Karadeniz Technical University, Trabzon 61080, Türkiye.
| | - Şengül Alpay Karaoğlu
- Department of Biology, Faculty of Science, Recep Tayyip Erdoğan University, Rize 53100, Türkiye
| | - Şeyma Suyabatmaz
- Department of Biology, Faculty of Science, Recep Tayyip Erdoğan University, Rize 53100, Türkiye
| | - Arif Bozdeveci
- Department of Biology, Faculty of Science, Recep Tayyip Erdoğan University, Rize 53100, Türkiye
| | - Gizem Tatar Yılmaz
- Department of Biostatistics and Medical Informatics, Faculty of Medicine, Karadeniz Technical University, Trabzon 61080, Türkiye; Karadeniz Technical University, Institute of Health Sciences, Department of Bioinformatics, 61080 Trabzon, Türkiye; Yılmaz Bilişim R&D Consulting Software Engineering and Services Trade Limited Company, 61081 Trabzon, Türkiye
| | - Nurettin Yaylı
- Department of Pharmacognosy, Faculty of Pharmacy, Karadeniz Technical University, Trabzon 61080, Türkiye
| | - Rahşan Akpınar
- Laboratory of Bee Diseases, Samsun Veterinary Control Institute, Samsun 55200, Türkiye
| | - Ayşegül Çopur Çiçek
- Department of Basic Medical Sciences, Faculty of Medicine, Istanbul Medipol University, Istanbul 34815, Türkiye
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24
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Ferraz MV, Adan WCS, Lima TE, Santos AJ, de Paula SO, Dhalia R, Wallau GL, Wade RC, Viana IF, Lins RD. Design of nanobody targeting SARS-CoV-2 spike glycoprotein using CDR-grafting assisted by molecular simulation and machine learning. PLoS Comput Biol 2025; 21:e1012921. [PMID: 40257976 PMCID: PMC12068729 DOI: 10.1371/journal.pcbi.1012921] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2024] [Revised: 05/12/2025] [Accepted: 02/26/2025] [Indexed: 04/23/2025] Open
Abstract
The design of proteins capable effectively binding to specific protein targets is crucial for developing therapies, diagnostics, and vaccine candidates for viral infections. Here, we introduce a complementarity-determining region (CDR) grafting approach for designing nanobodies (Nbs) that target specific epitopes, with the aid of computer simulation and machine learning. As a proof-of-concept, we designed, evaluated, and characterized a high-affinity Nb against the spike protein of SARS-CoV-2, the causative agent of the COVID-19 pandemic. The designed Nb, referred to as Nb Ab.2, was synthesized and displayed high-affinity for both the purified receptor-binding domain protein and to the virus-like particle, demonstrating affinities of 9 nM and 60 nM, respectively, as measured with microscale thermophoresis. Circular dichroism showed the designed protein's structural integrity and its proper folding, whereas molecular dynamics simulations provided insights into the internal dynamics of Nb Ab.2. This study shows that our computational pipeline can be used to efficiently design high-affinity Nbs with diagnostic and prophylactic potential, which can be tailored to tackle different viral targets.
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Affiliation(s)
- Matheus V.F. Ferraz
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
- Department of fundamental chemistry, Federal University of Pernambuco, Recife, Brazil
- Molecular and Cellular Modeling group, Heidelberg Institute for Theoretical Studies, Heidelberg, Germany
| | - W. Camilla S. Adan
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
- Department of fundamental chemistry, Federal University of Pernambuco, Recife, Brazil
| | - Tayná E. Lima
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
| | | | - Sérgio O. de Paula
- Department of General Biology, Federal University of Viçosa, Viçosa, Brazil
| | - Rafael Dhalia
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
| | - Gabriel L. Wallau
- Department of Entomology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
- Fiocruz Genomic Network, Oswaldo Cruz Foundation, Recife, Brazil
- Department of Arbovirology, Bernhard Nocht Institute for Tropical Medicine, WHO Collaborating Center for Arbovirus and Hemorrhagic Fever Reference and Research. National Reference Center for Tropical Infectious Diseases, Hamburg, Germany
| | - Rebecca C. Wade
- Molecular and Cellular Modeling group, Heidelberg Institute for Theoretical Studies, Heidelberg, Germany
- Center for Molecular Biology (ZMBH), DKFZ-ZMBH Alliance, Heidelberg University, Heidelberg, Germany
- Interdisciplinary Center for Scientific Computing (IWR), Heidelberg University, Heidelberg, Germany
| | - Isabelle F.T. Viana
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
- Fiocruz Genomic Network, Oswaldo Cruz Foundation, Recife, Brazil
| | - Roberto D. Lins
- Department of virology, Aggeu Magalhães Institute, Oswaldo Cruz Foundation, Recife, Brazil
- Fiocruz Genomic Network, Oswaldo Cruz Foundation, Recife, Brazil
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25
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Marinho LDSS, Andrade MCR, Lopes CADA, Coelho da Silva KVG, Gama E Souza KDM, Machado-Santos C. Immunohistochemical identification of ACE-2 (SARS-COV II entry mechanism) in the gastrointestinal tract, kidney and lung of rhesus monkeys (Macaca mulatta) and squirrel monkeys (Saimiri sciureus). Tissue Cell 2025; 93:102711. [PMID: 39787940 DOI: 10.1016/j.tice.2024.102711] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2024] [Revised: 11/15/2024] [Accepted: 12/27/2024] [Indexed: 01/12/2025]
Abstract
SARS-Cov-2 is a corona virus that causes COVID-19 disease, a viral infection responsible for the pandemic decreed by the World Health Organization in March 2020. Angiotensin-converting enzyme 2 (ACE-2) functions as the main receptor for SARS-Cov-2. The study aimed to detect the expression of ACE-2 in the gastrointestinal tract, kidney, and lung in the rhesus monkeys and squirrel monkeys. The sections from 18 rhesus monkey and 17 squirrel monkeys were incubated with rabbit polyclonal antibody to ACE2 (ab65863). In the lung of the rhesus monkeys, the presence of ACE-2 was noted in the bronchial mucosa of the respiratory epithelium. In the kidney, there was irregular in the proximal convoluted tubules. In the pyloric stomach, duodenum and in the large intestine it was observed on the surface of the lining epithelium. In the lung of the squirrel monkeys, this marking was present in both the ciliated cylindrical and goblet cell sof the bronchi. In the kidney light marking was observed along the surfasse of the cubic epithelium of the proximal convoluted tubules and in the renal glomerulus. No markings were observed throughout the stomach and intense staining was observed along the surfasse of the intestinal epithelium of the duodenum, jejunum and ileum, as well as in the intestinal glands. In our study, we can observe not able differences in the distribution of ACE2 between the two species of primates analysed. These differences must be considered in experimental studies on this disease, which continues to be a topic of notable importance for Public Health.
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Affiliation(s)
- Larissa Dos Santos Sebould Marinho
- Laboratory of Teaching and Research in Histology and Comparative Embryology (LEPHEC), Biomedical Institute, Fluminense Federal University, Niterói, RJ CEP 24210-130, Brazil
| | | | | | - Kassia Valéria Gomes Coelho da Silva
- Department of Pathology and Veterinary Clinic, Faculty of Veterinary, Fluminense Federal University, Vital Brazil/Santa Rosa, Niterói, RJ 24230-340, Brazil
| | - Kauet de Matos Gama E Souza
- Laboratory of Teaching and Research in Histology and Comparative Embryology (LEPHEC), Biomedical Institute, Fluminense Federal University, Niterói, RJ CEP 24210-130, Brazil
| | - Clarice Machado-Santos
- Laboratory of Teaching and Research in Histology and Comparative Embryology (LEPHEC), Biomedical Institute, Fluminense Federal University, Niterói, RJ CEP 24210-130, Brazil.
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26
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E Santo A, Reis A, Pinheiro AA, da Costa PI, Feliciano GT. Design of Mimetic Antibodies Targeting the SARS-CoV-2 Spike Glycoprotein Based on the GB1 Domain: A Molecular Simulation and Experimental Study. Biochemistry 2025; 64:1541-1549. [PMID: 40096593 PMCID: PMC11966750 DOI: 10.1021/acs.biochem.4c00671] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2024] [Revised: 02/18/2025] [Accepted: 03/12/2025] [Indexed: 03/19/2025]
Abstract
In the context of fast and significant technological transformations, it is natural for innovative artificial intelligence (AI) methods to emerge for the design of bioactive molecules. In this study, we demonstrated that the design of mimetic antibodies (MA) can be achieved using a combination of software and algorithms traditionally employed in molecular simulation. This combination, organized as a genetic algorithm (GA), has the potential to address one of the main challenges in the design of bioactive molecules: GA convergence occurs rapidly due to the careful selection of initial populations based on intermolecular interactions at antigenic surfaces. Experimental immunoenzymatic tests prove that the GA successfully optimized the molecular recognition capacity of one of the MA. One of the significant results of this study is the discovery of new structural motifs, which can be designed in an original and innovative way based on the MA structure itself, eliminating the need for preexisting databases. Through the GA developed in this study, we demonstrated the application of a new protocol capable of guiding experimental methods in the development of new bioactive molecules.
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Affiliation(s)
- Anderson
A. E Santo
- Institute
of Chemistry, São Paulo State University, Araraquara, SP 14800-900, Brazil
| | - Aline Reis
- School
of Pharmaceutical Sciences, São Paulo
State University, Araraquara, SP 14801-360, Brazil
| | - Anderson A. Pinheiro
- School
of Pharmaceutical Sciences, São Paulo
State University, Araraquara, SP 14801-360, Brazil
| | - Paulo I. da Costa
- School
of Pharmaceutical Sciences, São Paulo
State University, Araraquara, SP 14801-360, Brazil
| | - Gustavo T. Feliciano
- Institute
of Chemistry, São Paulo State University, Araraquara, SP 14800-900, Brazil
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27
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Sorotzky M, Raphael A, Breuer A, Odeh M, Gillis R, Gillis M, Shibli R, Fiszlinski J, Algur N, Magen S, Megged O, Schlesinger Y, Mendelovich J, Weiser G, Berliner E, Barak-Corren Y, Heiman E. Jerusalem's CoVID-19 Experience-The Effect of Ethnicity on Disease Prevalence and Adherence to Testing. J Racial Ethn Health Disparities 2025; 12:1315-1322. [PMID: 38457104 DOI: 10.1007/s40615-024-01965-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2023] [Revised: 02/22/2024] [Accepted: 03/01/2024] [Indexed: 03/09/2024]
Abstract
BACKGROUND The management of the SARS-CoV-2 pandemic depends amongst other factors on disease prevalence in the general population. The gap between the true rate of infection and the detected rate of infection may vary, especially between sub-groups of the population. Identifying subpopulations with high rates of undetected infection can guide authorities to direct resource distribution in order to improve health equity. METHODS A cross-sectional epidemiological survey was conducted between April and July 2021 in the Pediatric Emergency Department of the Shaare Zedek Medical Center, Jerusalem, Israel. We compared three categories: unconfirmed disease (UD), positive serology test result with no history of positive PCR; confirmed disease (CD), history of a positive PCR test result, regardless of serology test result; and no disease (ND), negative serology and no history of PCR. These categories were applied to local prevailing subpopulations: ultra-orthodox Jews (UO), National Religious Jews (NRJ), secular Jews (SJ), and Muslim Arabs (MA). RESULTS Comparing the different subpopulations groups, MAs and UOs had the greatest rate of confirmed or unconfirmed disease. MA had the highest rate of UD and UO had the highest rate of CD. UD significantly correlated with ethnicity, with a low prevalence in NRJ and SJ. UD was also associated with larger family size and housing density defined as family size per number of rooms. CONCLUSION This study highlights the effect of ethnicity on disease burden. These findings should serve to heighten awareness to disease burden in weaker populations and direct a suitable prevention program to each subpopulation's needs. Early awareness and possible intervention may lower morbidity and mortality.
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Affiliation(s)
- Michael Sorotzky
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel.
| | - Allon Raphael
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Adin Breuer
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Ma'aran Odeh
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Roni Gillis
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Michal Gillis
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Roaia Shibli
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Judith Fiszlinski
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
| | - Nurit Algur
- Clinical Endocrinology Laboratory, Shaare Zedek Medical Center, Jerusalem, Israel
| | - Sophie Magen
- Clinical Endocrinology Laboratory, Shaare Zedek Medical Center, Jerusalem, Israel
| | - Orli Megged
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Pediatric Infectious Diseases Unit, Shaare Zedek Medical Center, Jerusalem, Israel
- Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel
| | - Yechiel Schlesinger
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Pediatric Infectious Diseases Unit, Shaare Zedek Medical Center, Jerusalem, Israel
- Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel
| | - Joseph Mendelovich
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Pediatric Emergency Department, Shaare Zedek Medical Center, Jerusalem, Israel
| | - Giora Weiser
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel
- Pediatric Emergency Department, Shaare Zedek Medical Center, Jerusalem, Israel
| | - Elihay Berliner
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Pediatric Emergency Department, Shaare Zedek Medical Center, Jerusalem, Israel
| | - Yuval Barak-Corren
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Predictive Medicine Group, Boston Children's Hospital, Boston, USA
| | - Eyal Heiman
- Department of Pediatrics, Shaare Zedek Medical Center, 12 Shmuel Bait St, PO Box 3235, 9103102, Jerusalem, Israel
- Pediatric Emergency Department, Shaare Zedek Medical Center, Jerusalem, Israel
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28
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Nikbakht F, Heidarian Miri H, Mosafarkhani E, Sharifjafari F, Taghipour A. Role of COVID-19 infection status on the prediction of future infection: Immunity or susceptibility. PLoS One 2025; 20:e0317959. [PMID: 40138353 PMCID: PMC11940750 DOI: 10.1371/journal.pone.0317959] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/12/2024] [Accepted: 01/07/2025] [Indexed: 03/29/2025] Open
Abstract
BACKGROUND COVID-19 has rapidly spread around the world, and the duration of protective immunity against the virus remains unknown. Evidence suggests that patients with a confirmed COVID-19 infection may experience reinfection. The aim of this study is to determine the relationship between COVID-19 reinfection and previous infection history in the population covered by Mashhad University of Medical Sciences. METHODS This population-based, historical cohort study included all individuals with health records at the health service centers of Mashhad University of Medical Sciences who underwent PCR testing during the study period (April 1, 2020, up to February 19, 2022). The data were analyzed by calculating the infection rate in both PCR-positive and negative individuals, and estimating the adjusted rate ratio using Poisson regression. RESULTS The results of this study in the entire population showed that the incidence rate in people with a history of primary COVID-19 infection was 13% higher than that in people who had no history of this disease. However, in the group that received the vaccine prior to the first PCR test, the incidence rate was lower among individuals with a positive first test result (IRR = 0.71) compared to those with a negative first test result. CONCLUSION The study reveals that prior COVID-19 infection does not ensure immunity and may increase the risk of reinfection, particularly among men and younger individuals. Vaccination appears to complicate this dynamic, as those with multiple vaccine doses showed higher reinfection rates compared to those with fewer doses. These findings highlight the need for ongoing research and tailored public health strategies to address the complexities of COVID-19 immunity and reinfection.
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Affiliation(s)
- Fateme Nikbakht
- Department of Epidemiology, School of Health, Mashhad University of Medical Sciences, Mashhad, Iran
| | | | - Ehsan Mosafarkhani
- Department of Epidemiology, School of Health, Mashhad University of Medical Sciences, Mashhad, Iran
- Management and Social Determinants of Health Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
| | | | - Ali Taghipour
- Social Determinants of Health Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
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29
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Garcia-Carretero R, Ordoñez-Garcia M, Rodriguez-Gonzalez M, Barquero-Perez O, Gil-Prieto R, Gil-de-Miguel A. Nationwide study of COVID-19 outcomes in hematologic patients following bone marrow transplantation. Sci Rep 2025; 15:10506. [PMID: 40140426 PMCID: PMC11947223 DOI: 10.1038/s41598-025-95246-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/28/2024] [Accepted: 03/19/2025] [Indexed: 03/28/2025] Open
Abstract
Background Patients with hematological malignancies (HMs), particularly those who have undergone bone marrow or hematopoietic stem cell transplantation (HSCT), are at greater risk for morbidity and mortality due to immunosuppression. The COVID-19 pandemic exacerbated these vulnerabilities in HM patients, although comprehensive data specifically on HSCT recipients are limited. Objective This study investigated the clinical and demographic profiles of HSCT recipients hospitalized with COVID-19 in Spain. We also identified factors associated with in-hospital mortality in HSCT patients. Methods We conducted a nationwide, retrospective analysis using data from the Spanish National Health System. We included hospitalized patients with HMs and COVID-19 infection from 2020 to 2022. We used descriptive statistics, multivariate logistic regression, and survival analyses to assess predictors of mortality. Results In total, 35,648 patients with HMs were included, of whom 2,324 (6.5%) had undergone HSCT. The in-hospital mortality rate for HSCT recipients was 13%, lower than the 20% observed in non-HSCT patients. Older age, dementia, acute leukemia, and solid tumors were independently associated with increased mortality. In spite of their immunosuppressed state, HSCT recipients experienced relatively favorable outcomes, suggesting partial immune recovery following transplantation. Conclusions HSCT recipients with COVID-19 present different clinical characteristics and mortality risks than non-recipients. These findings indicate the need for specific management strategies for this vulnerable population. Further research is needed to explore immunological recovery and the transplant-specific factors that may influence COVID-19 outcomes.
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Affiliation(s)
- Rafael Garcia-Carretero
- Department of Internal Medicine, Mostoles University Hospital, Rey Juan Carlos University, Madrid, Spain.
| | - Maria Ordoñez-Garcia
- Department of Hematology, Mostoles University Hospital, Rey Juan Carlos University, Madrid, Spain
| | - Maria Rodriguez-Gonzalez
- Department of Internal Medicine, Mostoles University Hospital, Rey Juan Carlos University, Madrid, Spain
| | - Oscar Barquero-Perez
- Department of Signal Theory and Communications and Telematics Systems and Computing, Rey Juan Carlos University, Madrid, Spain
| | - Ruth Gil-Prieto
- Department of Preventive Medicine and Public Health, Rey Juan Carlos University, Madrid, Spain
| | - Angel Gil-de-Miguel
- Department of Preventive Medicine and Public Health, Rey Juan Carlos University, Madrid, Spain
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30
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Wei C, Sing CW, Wan EYF, Cheung CL, Wong ICK, Lai FTT. Multimorbidity incidence following hospitalization for SARS-CoV-1 infection or influenza over two decades: a territory-wide retrospective cohort study. NPJ Prim Care Respir Med 2025; 35:18. [PMID: 40133400 PMCID: PMC11937326 DOI: 10.1038/s41533-025-00424-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2025] [Accepted: 03/13/2025] [Indexed: 03/27/2025] Open
Abstract
An infection of SARS-CoV-1, the causative agent of Severe Acute Respiratory Syndrome (SARS), may be followed by long-term clinical sequala. We hypothesized a greater 20-year multimorbidity incidence in people hospitalized for SARS-CoV-1 infection than those for influenza during similar periods. We conducted a retrospective cohort study using a territory-wide public healthcare database in Hong Kong. All patients aged ≥15 hospitalized for SARS in 2003 or influenza in 2002 or 2004 with no more than one of 30 listed chronic disease were included. Demographics, clinical history, and medication use were adjusted for in the inverse-probability-of-treatment-weighted Poisson regression analyses. We identified 1255 hospitalizations for SARS-CoV-1 infection and 687 hospitalizations for influenza. Overall crude multimorbidity incident rates were 1.5 per 100 person-years among SARS patients and 5.6 among influenza patients. Adjusted multimorbidity incidence rate ratio (IRR) was estimated at 0.78 [95% confidence interval (CI), 0.70-0.86) for SARS patients compared with influenza patients. Analysis by follow-up period shows a potentially greater risk among SARS patients in the first year of follow-up (IRR 1.33, 95% CI 0.97-1.84), with the risk in influenza patients increasing in subsequent years. Subgroup analyses by age and sex showed consistent results with the main analysis that SARS-CoV-1 infection was not followed by a higher incidence of multimorbidity than influenza. Notable differences in the patterns of multimorbidity were identified between the two arms. To conclude, we found no evidence of a higher multimorbidity incidence after hospitalization for SARS than for influenza over the long-term.
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Affiliation(s)
- Cuiling Wei
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
| | - Chor Wing Sing
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
| | - Eric Yuk Fai Wan
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
- Laboratory of Data Discovery for Health (D24H), Hong Kong Science Park, Sha Tin, Hong Kong SAR, China
- Department of Family Medicine and Primary Care, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
- Advanced Data Analytics for Medical Science (ADAMS) Limited, Hong Kong SAR, China
| | - Ching Lung Cheung
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
- Laboratory of Data Discovery for Health (D24H), Hong Kong Science Park, Sha Tin, Hong Kong SAR, China
- Advanced Data Analytics for Medical Science (ADAMS) Limited, Hong Kong SAR, China
| | - Ian Chi Kei Wong
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China
- Advanced Data Analytics for Medical Science (ADAMS) Limited, Hong Kong SAR, China
- Aston Pharmacy School, Aston University, Birmingham, England, UK
| | - Francisco Tsz Tsun Lai
- Centre for Safe Medication Practice and Research, Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China.
- Laboratory of Data Discovery for Health (D24H), Hong Kong Science Park, Sha Tin, Hong Kong SAR, China.
- Department of Family Medicine and Primary Care, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pok Fu Lam, Hong Kong SAR, China.
- Advanced Data Analytics for Medical Science (ADAMS) Limited, Hong Kong SAR, China.
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31
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Khatun O, Kaur S, Tripathi S. Anti-interferon armamentarium of human coronaviruses. Cell Mol Life Sci 2025; 82:116. [PMID: 40074984 PMCID: PMC11904029 DOI: 10.1007/s00018-025-05605-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/01/2024] [Revised: 12/15/2024] [Accepted: 01/23/2025] [Indexed: 03/14/2025]
Abstract
Cellular innate immune pathways are formidable barriers against viral invasion, creating an environment unfavorable for virus replication. Interferons (IFNs) play a crucial role in driving and regulating these cell-intrinsic innate antiviral mechanisms through the action of interferon-stimulated genes (ISGs). The host IFN response obstructs viral replication at every stage, prompting viruses to evolve various strategies to counteract or evade this response. Understanding the interplay between viral proteins and cell-intrinsic IFN-mediated immune mechanisms is essential for developing antiviral and anti-inflammatory strategies. Human coronaviruses (HCoVs), including SARS-CoV-2, MERS-CoV, SARS-CoV, and seasonal coronaviruses, encode a range of proteins that, through shared and distinct mechanisms, inhibit IFN-mediated innate immune responses. Compounding the issue, a dysregulated early IFN response can lead to a hyper-inflammatory immune reaction later in the infection, resulting in severe disease. This review provides a brief overview of HCoV replication and a detailed account of its interaction with host cellular innate immune pathways regulated by IFN.
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Affiliation(s)
- Oyahida Khatun
- Emerging Viral Pathogens Laboratory, Centre for Infectious Disease Research, Indian Institute of Science, Bengaluru, India
- Microbiology & Cell Biology Department, Biological Sciences Division, Indian Institute of Science, Bengaluru, India
| | - Sumandeep Kaur
- Emerging Viral Pathogens Laboratory, Centre for Infectious Disease Research, Indian Institute of Science, Bengaluru, India
- Microbiology & Cell Biology Department, Biological Sciences Division, Indian Institute of Science, Bengaluru, India
| | - Shashank Tripathi
- Emerging Viral Pathogens Laboratory, Centre for Infectious Disease Research, Indian Institute of Science, Bengaluru, India.
- Microbiology & Cell Biology Department, Biological Sciences Division, Indian Institute of Science, Bengaluru, India.
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32
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Lacasse É, Dubuc I, Gudimard L, Andrade ACDSP, Gravel A, Greffard K, Chamberland A, Oger C, Galano JM, Durand T, Philipe É, Blanchet MR, Bilodeau JF, Flamand L. Delayed viral clearance and altered inflammatory responses affect severity of SARS-CoV-2 infection in aged mice. Immun Ageing 2025; 22:11. [PMID: 40075368 PMCID: PMC11899864 DOI: 10.1186/s12979-025-00503-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2024] [Accepted: 02/17/2025] [Indexed: 03/14/2025]
Abstract
Epidemiological investigations consistently demonstrate an overrepresentation of the elderly in COVID-19 hospitalizations and fatalities, making the advanced age as a major predictor of disease severity. Despite this, a comprehensive understanding of the cellular and molecular mechanisms explaining how old age represents a major risk factor remain elusive. To investigate this, we compared SARS-CoV-2 infection outcomes in young adults (2 months) and geriatric (15-22 months) mice. Both groups of K18-ACE2 mice were intranasally infected with 500 TCID50 of SARS-CoV-2 Delta variant with analyses performed on days 3, 5, and 7 post-infection (DPI). Analyses included pulmonary cytokines, lung RNA-seq, viral loads, lipidomic profiles, and histological assessments, with a concurrent evaluation of the percentage of mice reaching humane endpoints. The findings unveiled notable differences, with aged mice exhibiting impaired viral clearance, reduced survival, and failure to recover weight loss due to infection. RNA-seq data suggested greater lung damage and reduced respiratory function in infected aged mice. Additionally, elderly-infected mice exhibited a deficient antiviral response characterized by reduced Th1-associated mediators (IFNγ, CCL2, CCL3, CXCL9) and diminished number of macrophages, NK cells, and T cells. Furthermore, mass-spectrometry analysis of the lung lipidome indicated altered expression of several lipids with immunomodulatory and pro-resolution effects in aged mice such as Resolvin, HOTrEs, and NeuroP, but also DiHOMEs-related ARDS. These findings indicate that aging affects antiviral immunity, leading to prolonged infection, greater lung damage, and poorer clinical outcomes. This underscores the potential efficacy of immunomodulatory treatments for elderly subjects experiencing symptoms of severe COVID-19.
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Affiliation(s)
- Émile Lacasse
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
- Département de Microbiologie, Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada
| | - Isabelle Dubuc
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
| | - Leslie Gudimard
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
| | - Ana Claudia Dos S P Andrade
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
| | - Annie Gravel
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
| | - Karine Greffard
- Axe Endocrinologie et Néphrologie, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
| | | | - Camille Oger
- Institut Des Biomolécules Max Mousseron, UMR 5247, Pôle Chimie Balard Recherche, Université de Montpellier, CNRS, ENSCM, Montpellier, France
| | - Jean-Marie Galano
- Institut Des Biomolécules Max Mousseron, UMR 5247, Pôle Chimie Balard Recherche, Université de Montpellier, CNRS, ENSCM, Montpellier, France
| | - Thierry Durand
- Institut Des Biomolécules Max Mousseron, UMR 5247, Pôle Chimie Balard Recherche, Université de Montpellier, CNRS, ENSCM, Montpellier, France
| | - Éric Philipe
- Département de Chirurgie, Faculté de Médecine, Université, Québec, QC, Canada
| | - Marie-Renée Blanchet
- Département de Médecine, Faculté de Médecine, Université, Québec, QC, Canada
- Centre de Recherche de L'Institut de Cardiologie de Québec, Université, Québec, QC, Canada
| | - Jean-François Bilodeau
- Axe Endocrinologie et Néphrologie, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada
- Département de Médecine, Faculté de Médecine, Université, Québec, QC, Canada
| | - Louis Flamand
- Axe Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier, Universitaire de Québec- Université Laval, Québec, QC, Canada.
- Département de Microbiologie, Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada.
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Aranda J, Oriol I, Vázquez N, Ramos K, Suárez RC, Feria L, Peñafiel J, Coloma A, Borjabad B, Clivillé R, Vacas M, Carratalà J. Long COVID in ARDS Survivors: Insights from a Two-Year-Follow-Up Study After the First Wave of the Pandemic. J Clin Med 2025; 14:1852. [PMID: 40142660 PMCID: PMC11942911 DOI: 10.3390/jcm14061852] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/17/2025] [Revised: 03/06/2025] [Accepted: 03/08/2025] [Indexed: 03/28/2025] Open
Abstract
Objectives: To compare the health status, exercise capacity, and health-related quality of life (HRQoL) in survivors of COVID-19-associated acute respiratory distress syndrome (ARDS) at 8, 12, and 24 months post-diagnosis. Methods: We conducted a prospective, single-center follow-up study embedded within a larger multicenter cohort of adults with COVID-19 who required hospital admission. Eligible participants underwent clinical interviews, physical examinations, chest radiography, and the 6-min walk test (6MWT). Standardized scales were used to assess post-traumatic stress disorder (PTSD), anxiety, depression, and HRQoL. Results: Out of 1295 patients with COVID-19, 365 developed ARDS, of whom 166 survived. After excluding deaths and loss to follow-up, 95 patients were monitored for 24 months. Over 60% of patients had persistent symptoms, though significant improvements were recorded in quality of life and physical recovery. More than 70% recovered their previous physical capacity, but 15% did not return to their usual lifestyle habits. Symptoms such as arthralgia and fatigue decreased, but cognitive issues, such as memory loss and insomnia, persisted. Radiological improvements were noted, although pulmonary function remained impaired. The prevalence of PTSD and anxiety decreased, while depression remained stable at around 30%. Conclusions: Long COVID continues to impose significant physical, mental, and social challenges. Symptoms like fatigue and anxiety have a profound impact on daily life. Strategies are urgently needed to help patients regain health and resume their normal lives.
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Affiliation(s)
- Judit Aranda
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
- Department of Internal Medicine, Consorci Sanitari Alt Penedès-Garraf, 08720 Vilafranca del Penedès, Spain
| | - Isabel Oriol
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
- Infectious Disease Department, Hospital Universitari de Bellvitge, 08907 L’Hospitalet de Llobregat, Spain
- Department of Research, Bellvitge Biomedical Research Institute (IDIBELL), 08907 L’Hospitalet de Llobregat, Spain
- Center for Biomedical Research in Infectious Diseases Network (CIBERINFEC), Instituto de Salud Carlos III, 28029 Madrid, Spain
- Clinical Science Department, Faculty of Medicine, University of Barcelona, 08007 Barcelona, Spain
| | - Núria Vázquez
- Department of Research, Bellvitge Biomedical Research Institute (IDIBELL), 08907 L’Hospitalet de Llobregat, Spain
- Center for Biomedical Research in Infectious Diseases Network (CIBERINFEC), Instituto de Salud Carlos III, 28029 Madrid, Spain
| | - Karim Ramos
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Romina Concepción Suárez
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Lucía Feria
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Judith Peñafiel
- Statistics Advisory Service, Bellvitge Biomedical Research Institute (IDIBELL), 08907 L’Hospitalet de Llobregat, Spain
| | - Ana Coloma
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Beatriz Borjabad
- Department of Internal Medicine, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Raquel Clivillé
- Department of Microbiology, CLILAB Diagnòstics, 08720 Barcelona, Spain
| | - Montserrat Vacas
- Department of Psychiatry and Psychology, Complex Hospitalari Moisès Broggi, 08970 Sant Joan Despí, Spain
| | - Jordi Carratalà
- Infectious Disease Department, Hospital Universitari de Bellvitge, 08907 L’Hospitalet de Llobregat, Spain
- Department of Research, Bellvitge Biomedical Research Institute (IDIBELL), 08907 L’Hospitalet de Llobregat, Spain
- Center for Biomedical Research in Infectious Diseases Network (CIBERINFEC), Instituto de Salud Carlos III, 28029 Madrid, Spain
- Clinical Science Department, Faculty of Medicine, University of Barcelona, 08007 Barcelona, Spain
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de Souza AMLB, de Araújo EF, Junior NC, Raimundo ACS, Pereira AC, de Castro Meneghim M. Association between SARS-CoV-2 and stroke: perspectives from a metaumbrella-review. BMC Neurol 2025; 25:97. [PMID: 40055630 PMCID: PMC11887298 DOI: 10.1186/s12883-025-04041-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2024] [Accepted: 01/15/2025] [Indexed: 05/13/2025] Open
Abstract
In the face of the global COVID-19 pandemic, the need arose to investigate potential complications associated with SARS-CoV-2, including the risk of stroke.ObjectiveThis study aimed to verify the association between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the risk of stroke on the basis of systematic reviews and meta-analyses to assess the inclusion of the virus as a new risk factor for cerebrovascular diseases.MethodsA metaumbrella study was conducted, which included 34 systematic reviews, of which 4 were selected for the final analysis on the basis of methodological quality and consistency. The analysis aggregated the results of 70 primary studies, considering different stroke subtypes and outcomes associated with COVID-19. Study heterogeneity was assessed via the I2 index, and significance bias was verified via Egger's test.ResultsCOVID-19 severity was significantly associated with an increased risk of stroke (eOR = 2.48; 95% CI: 1.55-3.95), particularly ischemic stroke (eOR = 1.76; 95% CI: 1.11-2.80) and hemorrhagic stroke (eOR = 3.86; 95% CI: 1.79-8.33). Additionally, patients with cerebrovascular comorbidities had higher mortality (eOR = 2.48; 95% CI: 2.48-19.63), as did those who had previously suffered a stroke (eOR = 6.08; 95% CI: 3.73-9.91).ConclusionThe association between SARS-CoV-2 and stroke incidence was consistent and significant, suggesting that COVID-19 should be considered a new risk factor for cerebrovascular diseases. However, the high heterogeneity among the studies analyzed reinforces the need for further research to consolidate this relationship.
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Affiliation(s)
- Andreza Maria Luzia Baldo de Souza
- Faculdade de Odontologia de Piracicaba/FOP, departamento de Ciências da Saúde E Odontologia Infantil, Universidade Estadual de Campinas/UNICAMP, Avenida Limeira 901, Bairro Areião, Piracicaba-SP, CEP13414903, Brazil.
| | - Enoque Fernandes de Araújo
- Faculdade de Odontologia de Piracicaba/FOP, departamento de Ciências da Saúde E Odontologia Infantil, Universidade Estadual de Campinas/UNICAMP, Avenida Limeira 901, Bairro Areião, Piracicaba-SP, CEP13414903, Brazil
| | | | - Augusto Cesar Sousa Raimundo
- Faculdade de Odontologia de Piracicaba/FOP, departamento de Ciências da Saúde E Odontologia Infantil, Universidade Estadual de Campinas/UNICAMP, Avenida Limeira 901, Bairro Areião, Piracicaba-SP, CEP13414903, Brazil
| | - Antonio Carlos Pereira
- Faculdade de Odontologia de Piracicaba/FOP, departamento de Ciências da Saúde E Odontologia Infantil, Universidade Estadual de Campinas/UNICAMP, Avenida Limeira 901, Bairro Areião, Piracicaba-SP, CEP13414903, Brazil
| | - Marcelo de Castro Meneghim
- Faculdade de Odontologia de Piracicaba/FOP, departamento de Ciências da Saúde E Odontologia Infantil, Universidade Estadual de Campinas/UNICAMP, Avenida Limeira 901, Bairro Areião, Piracicaba-SP, CEP13414903, Brazil
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Wu M, Wang K, Wang H, Yan H, Wu S, Yang G, Li Y, Che Y, Jiang J. Mycophenolate mofetil exerts broad-spectrum antiviral activity against coronaviruses including SARS-CoV-2. Virol J 2025; 22:56. [PMID: 40038695 PMCID: PMC11877706 DOI: 10.1186/s12985-025-02673-2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2024] [Accepted: 02/17/2025] [Indexed: 03/06/2025] Open
Abstract
BACKGROUND New anti-coronavirus drugs are continuously being developed to address the serious long-term challenge posed by numerous SARS-CoV-2 variants. The clinical immunosuppressants mycophenolate mofetil (MMF) and mycophenolic acid (MPA) have been reported to have anti-coronavirus activities. However, systematic studies have not been conducted to evaluate their activities and mechanisms against pan-coronaviruses, including SARS-CoV-2. METHODS The antiviral effect of MMF and MPA was determined by qRT-PCR assay, Western blotting, and immunofluorescence assay. The IMPDH inhibition effect of MMF was determined by cellular thermal shift assay and Western blotting. RESULTS We showed that MMF and MPA had broad-spectrum inhibitory effect against coronavirus, including HCoV-229E, HCoV-OC43, and SARS-CoV-2 ancestral strain and its variants. In terms of characteristics, MMF acted in the early stages of viral infection and inhibited viral replication by blocking purine nucleotide synthesis through interaction with inosine-5'-monophosphate dehydrogenase (IMPDH). Therefore, the antiviral effect of MMF can be reversed by exogenous guanosine. Additionally, MMF in combination with molnupiravir, GC376 or E64d showed synergistic antiviral effects. CONCLUSION MMF and MPA exerted broad-spectrum anti-coronavirus effects by inhibiting IMPDH activity. MMF had a synergistic antiviral effect when combined with other drugs, showing its potential clinical antiviral applications.
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Affiliation(s)
- Mengyuan Wu
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | - Kun Wang
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | - Huiqiang Wang
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | - Haiyan Yan
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | - Shuo Wu
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
- State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China
| | - Ge Yang
- CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Technology and Application for Anti-Infective New Drugs Research and Development, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
| | - Yuhuan Li
- State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
| | - Yongsheng Che
- State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
| | - Jiandong Jiang
- State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
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Li J, Jia H, Chen C, An S, Yu J, Yuan J, Long Y, Li M. Congested heptacyclic meroterpenoids with anti-SARS-CoV-2 and anti-inflammatory activities from mangrove endophytic fungus Talaromyces amestolkiae SCNU-F0041. PHYTOCHEMISTRY 2025; 231:114344. [PMID: 39613279 DOI: 10.1016/j.phytochem.2024.114344] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/17/2024] [Revised: 11/22/2024] [Accepted: 11/26/2024] [Indexed: 12/01/2024]
Abstract
Under the guidance of LC‒MS/MS-based molecular networking, three previously undescribed berkeleyacetal type meroterpenoids, amestolknoids A-C (1-3), together with ten known analogues (4-13) were isolated and identified from the mangrove endophytic fungus Talaromyces amestolkiae SCNU-F0041. Amestolknoids A (1) and B (2) are unprecedentedly congested 6/7/6/5/5/5/5 heptacyclic scaffolds characterized by two chiral spiroketal centers. Amestolknoids A (1) and C (3) represent the first examples of chlorinated berkeleyacetal type meroterpenoids. Their structures were established by extensive spectroscopic analyses and single crystal X-ray diffraction. Bioassays revealed that amestolknoid C (3) exhibited excellent antiviral activity against SARS-CoV-2 with an EC50 value of 2.50 μM and strong inhibitory effects on nitric oxide production in lipopolysaccharide-activated RAW 264.7 cells with an IC50 value of 4.10 μM. Compounds 2, 4, 6, 9 and 11 showed moderate to good anti-inflammatory activities with IC50 values of 41.78, 3.91, 43.52, 20.79, and 33.26 μM, respectively.
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Affiliation(s)
- Jialin Li
- School of Chemistry, Guangzhou Key Laboratory of Analytical Chemistry for Biomedicine, South China Normal University, Guangzhou, 510006, China.
| | - Hao Jia
- School of Chemistry, Guangzhou Key Laboratory of Analytical Chemistry for Biomedicine, South China Normal University, Guangzhou, 510006, China
| | - Chen Chen
- School of Chemistry, Guangzhou Key Laboratory of Analytical Chemistry for Biomedicine, South China Normal University, Guangzhou, 510006, China
| | - Shu An
- Key Laboratory of Tropical Disease Control, Ministry of Education, Sun Yat-sen University, Guangzhou, China; Department of Laboratory Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
| | - Jianchen Yu
- Key Laboratory of Tropical Disease Control (Sun Yat-sen University), Ministry of Education, Guangzhou, 510080, China; Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, China.
| | - Jie Yuan
- Key Laboratory of Tropical Disease Control (Sun Yat-sen University), Ministry of Education, Guangzhou, 510080, China; Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510080, China.
| | - Yuhua Long
- School of Chemistry, Guangzhou Key Laboratory of Analytical Chemistry for Biomedicine, South China Normal University, Guangzhou, 510006, China.
| | - Mengfeng Li
- Key Laboratory of Tropical Disease Control (Sun Yat-sen University), Ministry of Education, Guangzhou, 510080, China; Cancer Institute, Southern Medical University, Guangzhou, 510515, China
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Mahmood J, Ul Haque MI, Gul M, Ayub A, Ansari FA, Ahmad W. Early Identification of Severe COVID-19 Cases and the Need for ICU Care Based on Clinical and Laboratory Risk Factors. Cureus 2025; 17:e80611. [PMID: 40230780 PMCID: PMC11995811 DOI: 10.7759/cureus.80611] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 03/12/2025] [Indexed: 04/16/2025] Open
Abstract
Background and objective Treatment in ICUs became extremely difficult due to the growing number of coronavirus disease 2019 (COVID-19) patients at the height of the pandemic. Consequently, prompt patient triage depends on the early categorization of severe cases in such scenarios. This study aimed to provide an evidence-based strategy to ensure the best use of resources by triaging patients based on objective risk factors. Methods This retrospective observational study comprised 500 inpatients (>age 18 years) who were hospitalized between March 20 and April 19, 2020, at the Khyber Teaching Hospital (KTH) and Hayatabad Medical Complex (HMC) in Peshawar, Pakistan. The clinical, laboratory, and radiological parameters were assessed. Real-time polymerase chain reaction (RT-PCR) findings were used to confirm the diagnosis of COVID-19. Results A total of 19 potential clinical and laboratory risk factors associated with ICU admissions were identified. At least one comorbidity among chronic lung disease, cardiovascular disease (CVD), and diabetes was the factor with the strongest association with ICU admission with a univariable odds ratio (OR) of over 27, followed by renal disease and other COVID-19 sequelae such as diarrhea, respiratory rate (>24 breaths/minute), and positive RT-PCR (vs. negative) with an univariable OR between 9 and 15. Furthermore, a multivariate logistic regression model was further developed with five risk factors, including comorbidity, presence of chronic lung disease, presence of diabetes, and RT-PCR (positive vs. negative), male sex (vs. female), and older age (65.0-80.5 years), suggesting a good fit of the model to the data shown by the area under the receiver operator characteristic curve (AUC) of 0.943 (95% CI: 0.917, 0.969). Additionally, a chest CT scan showed the typical COVID-19 pneumonia with pulmonary involvement of 30-40%, which was further evaluated by the COVID-19 Reporting and Data System (CO-RADS). The typical COVID-19 pneumonia was on a scale of four (15/25) or five (19/25) lung lesions. Conclusions Based on our findings, this approach could be used to screen the severe cases of COVID-19 patients and help them to be treated in ICUs on time while preventing others from unnecessarily using ICUs in the setting of limited medical resources, such as the outbreak of a pandemic.
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Affiliation(s)
- Jawad Mahmood
- Gastroenterology and Hepatology, Hayatabad Medical Complex, Peshawar, PAK
| | - Muhammad Izhar Ul Haque
- Department of Comparative Biomedical Sciences, College of Veterinary Medicine, University of Georgia, Athens, USA
| | - Maria Gul
- Gynecology, Ayub Teaching Hospital, Abbottabad, PAK
| | - Aliya Ayub
- Epidemiology and Public Health, Institute of Public Health and Social Sciences, Khyber Medical University, Peshawar, PAK
| | - Fawwad A Ansari
- Internal Medicine, Piedmont Athens Regional Medical Center, Athens, USA
| | - Wiqas Ahmad
- Gastroenterology and Hepatology, Hayatabad Medical Complex, Peshawar, PAK
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Bahojb Mahdavi SZ, Jebelli A, Aghbash PS, Baradaran B, Amini M, Oroojalian F, Pouladi N, Baghi HB, de la Guardia M, Mokhtarzadeh AA. A comprehensive overview on the crosstalk between microRNAs and viral pathogenesis and infection. Med Res Rev 2025; 45:349-425. [PMID: 39185567 PMCID: PMC11796338 DOI: 10.1002/med.22073] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2021] [Revised: 04/11/2023] [Accepted: 08/04/2024] [Indexed: 08/27/2024]
Abstract
Infections caused by viruses as the smallest infectious agents, pose a major threat to global public health. Viral infections utilize different host mechanisms to facilitate their own propagation and pathogenesis. MicroRNAs (miRNAs), as small noncoding RNA molecules, play important regulatory roles in different diseases, including viral infections. They can promote or inhibit viral infection and have a pro-viral or antiviral role. Also, viral infections can modulate the expression of host miRNAs. Furthermore, viruses from different families evade the host immune response by producing their own miRNAs called viral miRNAs (v-miRNAs). Understanding the replication cycle of viruses and their relation with host miRNAs and v-miRNAs can help to find new treatments against viral infections. In this review, we aim to outline the structure, genome, and replication cycle of various viruses including hepatitis B, hepatitis C, influenza A virus, coronavirus, human immunodeficiency virus, human papillomavirus, herpes simplex virus, Epstein-Barr virus, Dengue virus, Zika virus, and Ebola virus. We also discuss the role of different host miRNAs and v-miRNAs and their role in the pathogenesis of these viral infections.
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Affiliation(s)
- Seyedeh Zahra Bahojb Mahdavi
- Department of Biology, Faculty of Basic SciencesAzarbaijan Shahid Madani UniversityTabrizIran
- Immunology Research CenterTabriz University of Medical SciencesTabrizIran
| | - Asiyeh Jebelli
- Department of Biological Science, Faculty of Basic ScienceHigher Education Institute of Rab‐RashidTabrizIran
- Tuberculosis and Lung Diseases Research CenterTabriz University of Medical SciencesTabrizIran
| | | | - Behzad Baradaran
- Immunology Research CenterTabriz University of Medical SciencesTabrizIran
| | - Mohammad Amini
- Immunology Research CenterTabriz University of Medical SciencesTabrizIran
| | - Fatemeh Oroojalian
- Department of Advanced Sciences and Technologies in Medicine, School of MedicineNorth Khorasan University of Medical SciencesBojnurdIran
| | - Nasser Pouladi
- Department of Biology, Faculty of Basic SciencesAzarbaijan Shahid Madani UniversityTabrizIran
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Maliha ST, Fatemi R, Akter M, Zheng Q, Araf Y, Tabassum T, Munif MR, Saha S, Xue M, Wang H, Zheng C, Hossain MG. Exploring the dynamics of SARS-CoV-2 and HIV Co-infection: Mutation risks, therapeutic efficacy, and future variant prevention. Diagn Microbiol Infect Dis 2025; 111:116707. [PMID: 39854809 DOI: 10.1016/j.diagmicrobio.2025.116707] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2024] [Revised: 01/19/2025] [Accepted: 01/20/2025] [Indexed: 01/27/2025]
Abstract
High mutation rates in SARS-CoV-2, particularly among immunocompromised patients living with HIV, continue to complicate the current COVID-19 pandemic. The threshold for severe COVID-19 and a greater risk of mortality have increased in many immunocompromised individuals due to a weakened immune system. Low CD4+ T-cell counts in people living with both HIV and COVID-19 lead to prolonged disease duration and, therefore, an increased likelihood of viral infection with SARS-CoV-2 mutations in such individuals. These mutations could decrease the efficiency of ongoing vaccines and cause new outbreaks. Recently, the rise of new mutations in this patient population has created increasing concern; however, few data are currently available on the direct association of HIV infection with SARS-CoV-2 mutations. This review highlights the implications of SARS-CoV-2 and HIV co-infection, highlighting the need for extra caution and monitoring of the immune-compromised population during a pandemic. Access to HIV care and COVID-19 treatments, careful surveillance, and adapted health strategies are key to reducing risks and protecting these populations. Further research is required to elucidate the dynamics of mutations and develop intervention methods to manage COVID-19 among immunocompromised patients.
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Affiliation(s)
- Sumaiya Tasnim Maliha
- Biotechnology Program, Department of Mathematics and Natural Sciences, School of Data and Sciences, BRAC University, Dhaka, Bangladesh
| | - Rabeya Fatemi
- Department of Genetic Engineering and Biotechnology, East-West University, Dhaka 1212, Bangladesh
| | - Marjana Akter
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
| | - Qingcong Zheng
- Department of Spinal Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China
| | - Yusha Araf
- Department of Biotechnology, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
| | - Tahani Tabassum
- Biotechnology Program, Department of Mathematics and Natural Sciences, School of Data and Sciences, BRAC University, Dhaka, Bangladesh
| | - Mohammad Raguib Munif
- Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
| | - Sukumar Saha
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh
| | - Mengzhou Xue
- Department of Cerebrovascular Diseases, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, 450001, China.
| | - Huiqing Wang
- Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
| | - Chunfu Zheng
- Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB, Canada.
| | - Md Golzar Hossain
- Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh.
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Ghammaz H, Melloul M, Mbarki A, Hemlali M, Chouati T, Annaz HE, Touil N, Elouennass M, Ennibi K, Fahime EE. Genomic evolution of SARS-CoV-2 in Morocco: Insights from whole genome sequences collected from 2020 to 2024. Virus Res 2025; 353:199530. [PMID: 39864629 PMCID: PMC11841124 DOI: 10.1016/j.virusres.2025.199530] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/19/2024] [Revised: 10/08/2024] [Accepted: 01/21/2025] [Indexed: 01/28/2025]
Abstract
This study investigates the evolution and genetic diversity of SARS-CoV-2 strains circulating in Morocco to track the spread, clade distributions and mutations of the virus across various regions from February 2020 to June 2024. The genome sequences were retrieved from the GISAID database. A total of 2630 SARS-CoV-2 genome sequences were analyzed using bioinformatic tools such as Nextclade, followed by phylogenetic and statistical analyses. The study highlights the predominance of the GRA clade (Omicron variant) since November 2021, while clades such as G, GH, GR, and GRY were identified earlier. The GRA clade exhibited the highest number of non-synonymous mutations, particularly in the Spike (S) gene, suggesting strong evolutionary pressure. The correlation analysis between structural and non-structural proteins revealed key interactions between S and NSP5, providing insights into the viral replication and assembly processes. This work gives new insights to the dynamics of SARS-CoV-2 in Morocco and underscores the importance of ongoing genomic surveillance to respond to emerging variants and potential future outbreaks.
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Affiliation(s)
- Hamza Ghammaz
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Genomic Centre for Human Pathologies (GENOPATH), Neuroscience and Neurogenetics Research Team, Faculty of Medicine and Pharmacy, University Mohammed V, Rabat, Morocco
| | - Marouane Melloul
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Microbiology and Molecular Biology Team, Center of Plant and Microbial Biotechnology, Biodiversity, and Environment, Faculty of Sciences, Mohammed V University of Rabat, Rabat, Morocco
| | - Ahlam Mbarki
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Microbiology and Molecular Biology Team, Center of Plant and Microbial Biotechnology, Biodiversity, and Environment, Faculty of Sciences, Mohammed V University of Rabat, Rabat, Morocco
| | - Mouhssine Hemlali
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Genomic Centre for Human Pathologies (GENOPATH), Neuroscience and Neurogenetics Research Team, Faculty of Medicine and Pharmacy, University Mohammed V, Rabat, Morocco
| | - Taha Chouati
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Genomic Centre for Human Pathologies (GENOPATH), Neuroscience and Neurogenetics Research Team, Faculty of Medicine and Pharmacy, University Mohammed V, Rabat, Morocco
| | - Hicham El Annaz
- Cell Culture Unit, Center of Virology, Infectious and Tropical Diseases, the Mohammed V Military Training Hospital, Rabat, Morocco
| | - Nadia Touil
- Cell Culture Unit, Center of Virology, Infectious and Tropical Diseases, the Mohammed V Military Training Hospital, Rabat, Morocco
| | - Mostafa Elouennass
- Department of Bacteriology, Mohammed V Military Teaching Hospital, Rabat, Morocco
| | - Khalid Ennibi
- Cell Culture Unit, Center of Virology, Infectious and Tropical Diseases, the Mohammed V Military Training Hospital, Rabat, Morocco; Immunopathology Research Team (ERIP), Faculty of Medicine of Pharmacy, University Mohammed V, Rabat, Morocco
| | - Elmostafa El Fahime
- Molecular Biology and Functional Genomics Platform, National Centre for Scientific and Technical Research (CNRST), Rabat, Morocco; Genomic Centre for Human Pathologies (GENOPATH), Neuroscience and Neurogenetics Research Team, Faculty of Medicine and Pharmacy, University Mohammed V, Rabat, Morocco; University Mohammed VI of Science and Health (UM6SS), Casablanca, Morocco; Genomics and Molecular Biology, Mohammed Vi Center for Research and Innovation (CM6RI), Rabat, Morocco.
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Jivapetthai A, Arunmanee W, Pornputtapong N. Quality by design for transient RBD-Fc fusion protein production in Chinese hamster ovary cells. BIOTECHNOLOGY REPORTS (AMSTERDAM, NETHERLANDS) 2025; 45:e00882. [PMID: 40034964 PMCID: PMC11872631 DOI: 10.1016/j.btre.2025.e00882] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 09/08/2024] [Revised: 01/13/2025] [Accepted: 02/07/2025] [Indexed: 03/05/2025]
Abstract
Quality by design (QbD) is applied to the upstream process to maximize the RBD-Fc fusion protein production in CHO cells. The three factors (culture duration, temperature, and polyethyleneimine to plasmid DNA (PEI-Max/pDNA) ratio) were identified as critical process attributes based on risk analysis (FMEA) and further optimized by response surface to maximize the protein yields. Using a Box-Behnken design, the optimal conditions for RBD-Fc production were determined to be a culture duration of 5 days, a culture temperature of 34.4 °C, and a PEI-Max/pDNA ratio of 4.2:1 (w/w) with a predictive value of 48 mg/L (desirability of 92.8 %). The PEI-Max/pDNA ratio and its interaction with culture duration to express the highest yield (47.78 ± 2.30 mg/l). In addition, the purified CHO-produced RBD-Fc fusion protein was highly pure and strongly bound to its receptor, ACE2. Our finding demonstrated that the QBD tools can identify the critical parameters to facilitate scaling-up production.
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Affiliation(s)
- Araya Jivapetthai
- Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand
| | - Wanatchaporn Arunmanee
- Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand
- Center of Excellence in Cancer Cell and Molecular Biology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand
| | - Natapol Pornputtapong
- Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand
- Center of Excellence in Systems Biology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
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Marin L, de Miranda LS, Carvalho VHS, Voigt MEF, Martire JPL, Nunes MRT, Slhessarenko RD. Phylogeography of SARS-CoV-2 Omicron sublineages detected in asymptomatic blood donors during third epidemiological wave in Mato Grosso, Midwestern Brazil. Diagn Microbiol Infect Dis 2025; 111:116693. [PMID: 39864307 DOI: 10.1016/j.diagmicrobio.2025.116693] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/25/2024] [Revised: 01/13/2025] [Accepted: 01/15/2025] [Indexed: 01/28/2025]
Abstract
Emerging infectious disease agents represent pathogens that may evade current screening protocols while posing significant transfusion transmission risks regionally. This study investigated the prevalence of SARS-CoV-2 and other respiratory viruses among 633 blood donors at the MT-Hemocentro from November 2021 to February 2023. Nucleic acid obtained from nasopharyngeal swabs were tested by RT-qPCR for SARS-CoV-2, RSV, FLU-A, and FLU-B. Serum from positive samples was also tested for nucleic acid. The prevalence of SARS-CoV-2 was 6.48 % (41/633); 2 of the 41 blood donors had SARSCoV-2 detectable in their serum. All positive samples were collected between January 2022 and March 2023, coinciding with the third epidemic wave in Brazil; 97.6 % of these SARS-CoV-2-positive donors were vaccinated with at least two doses. SARS-CoV-2 genomes recovered from six nasopharyngeal samples were classified into BA.1.1.1, BA.1.14.1, BA.2, BA.5.1, BA.5.2.1 sublineages. Phylogeographic analysis across Brazil's five regions revealed that the Northeast acted as the main exporter of Omicron sublineages, while the South and Southeast regions were more frequently importers.
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Affiliation(s)
- Leonardo Marin
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil; MT-Hemocentro, Secretaria Estadual de Saúde, Governo do Estado de Mato Grosso, Cuiabá, MT, USA
| | - Lucas Santos de Miranda
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil
| | - Victor Hugo Silveira Carvalho
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil
| | - Maria Eduarda Fantacholi Voigt
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil
| | - João Pedro Lopes Martire
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil
| | - Márcio Roberto Teixeira Nunes
- Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, Pará, Brazil
| | - Renata Dezengrini Slhessarenko
- Laboratório de Virologia, Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso, Brazil.
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Wong KT, Hooi YT, Tan SH, Ong KC. Emerging and re-emerging viral infections of the central nervous system in Australasia and beyond. Pathology 2025; 57:230-240. [PMID: 39799084 DOI: 10.1016/j.pathol.2024.11.003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2024] [Revised: 11/24/2024] [Accepted: 11/27/2024] [Indexed: 01/15/2025]
Abstract
Viral infections of the central nervous system (CNS) have been emerging and re-emerging worldwide, and the Australasia region has not been spared. Enterovirus A71 and enterovirus D68, both human enteroviruses, are likely to replace the soon-to-be eradicated poliovirus to cause global outbreaks associated with neurological disease. Although prevalent elsewhere, the newly emergent orthoflavivirus, Japanese encephalitis virus (genotype IV), caused human infections in Australia in 2021, and almost certainly will continue to do so because of spillovers from the natural animal host-vector life cycle endemic in the country. Another orthoflavivirus, Murray Valley encephalitis virus, has re-emerged in Australia. The Hendra henipavirus together with Nipah henipavirus are listed as high-risk pathogens by the World Health Organization because both can cause lethal encephalitis. The former remains a health threat in Australasia because bats may still be able to spread the infection to unvaccinated Australian horses and other animals acting as intermediate hosts, and thence to humans. The global COVID-19 pandemic, caused by the emerging severe acute respiratory syndrome coronavirus-2, a virus transmitted from animals to humans that was first described and first arose in China, is associated with acute and long-lasting CNS pathology. Fortunately, the pathology and pathogenesis of these important neurotropic viruses are now better understood, leading to better management protocols and prevention strategies. Pathologists are in a unique position to contribute to the diagnosis and advancement in our knowledge of infectious diseases. This review summarises some of the current knowledge about a few important emerging and re-emerging CNS infections in Australasia and beyond.
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Affiliation(s)
- Kum Thong Wong
- Department of Pathology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Selangor, Malaysia.
| | - Yuan Teng Hooi
- Infection and Immunity Research Strength, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Selangor, Malaysia
| | - Soon Hao Tan
- Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
| | - Kien Chai Ong
- Department of Biomedical Science, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia
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Didkowska A, Martín-Santander V, Wojciechowska M, Olech W, Anusz K, Fernández A, Davies JE, Gómez Á, Peña-Fresneda N, Arias M, Lacasta D, Ortín A, Pérez MD, Villanueva-Saz S, Marteles D. Presence of anti-SARS-CoV-2 antibodies in European bison (Bison bonasus) in Poland, 2019-2023. BMC Vet Res 2025; 21:120. [PMID: 40022124 PMCID: PMC11869555 DOI: 10.1186/s12917-025-04593-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2024] [Accepted: 02/11/2025] [Indexed: 03/03/2025] Open
Abstract
BACKGROUND The origin of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains unknown. However, it is likely that the virus spillover occurred from an animal reservoir to humans. Identifying animal species susceptible to SARS-CoV-2 is crucial for understanding cross-species transmission to humans. This study distinguishes itself by focusing on the susceptibility of the European bison (Bison bonasus), an endangered species, to SARS-CoV-2. The objective of this study was to investigate the occurrence of SARS-CoV-2 antibodies in a substantial number (n = 238) of both free-living and captive Polish European bison using an in-house ELISA method and virus neutralization test (VNT). RESULTS The seroprevalence of SARS-CoV-2 infection was found to be 1.29% (3/232). None of the seropositive European bison tested positive in the virus neutralization test. All seropositive animals were part of captive herds. CONCLUSIONS This study represents the first report of SARS-CoV-2 seroprevalence in both free-ranging and captive European bison in Poland. Based on these findings, the European bison appears to be a less susceptible species to SARS-CoV-2. The most probable route of transmission was from humans to European bison, as all seropositive animals belonged to captive herds with contact with indirect human sources, such as tourists and keepers.
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Affiliation(s)
- Anna Didkowska
- Department of Food Hygiene and Public Health Protection, Institute of Veterinary Medicine, Warsaw University of Life Sciences (SGGW), Nowoursynowska 166, Warsaw, 02-787, Poland.
| | - Víctor Martín-Santander
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
| | - Marlena Wojciechowska
- Department of Animal Genetics and Conservation, Warsaw University of Life Sciences (SGGW), Ciszewskiego 8, Warsaw, 02-786, Poland
| | - Wanda Olech
- Department of Animal Genetics and Conservation, Warsaw University of Life Sciences (SGGW), Ciszewskiego 8, Warsaw, 02-786, Poland
| | - Krzysztof Anusz
- Department of Food Hygiene and Public Health Protection, Institute of Veterinary Medicine, Warsaw University of Life Sciences (SGGW), Nowoursynowska 166, Warsaw, 02-787, Poland
| | - Antonio Fernández
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain
| | - Janine E Davies
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
| | - Álex Gómez
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain
| | | | - Maykel Arias
- Aragon Health Research Institute (IIS Aragón), Zaragoza, 50009, Spain
- CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III, Madrid, 28029, Spain
| | - Delia Lacasta
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain
| | - Aurora Ortín
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain
| | - María Dolores Pérez
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain
- Department of Animal Production and Sciences of the Food, Veterinary Faculty, University of Zaragoza, Zaragoza, Spain
| | - Sergio Villanueva-Saz
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain.
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain.
- Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Zaragoza, 50013, Spain.
| | - Diana Marteles
- Department of Animal Pathology, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
- Clinical Immunology Laboratory, Veterinary Faculty, University of Zaragoza, Zaragoza, 50013, Spain
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Luo S, Huang X, Chen S, Li J, Wu H, He Y, Zhou L, Liu B, Feng J. The Gut Microbiota of the Greater Horseshoe Bat Confers Rapidly Corresponding Immune Cells in Mice. Animals (Basel) 2025; 15:685. [PMID: 40075967 PMCID: PMC11899282 DOI: 10.3390/ani15050685] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2025] [Revised: 02/19/2025] [Accepted: 02/24/2025] [Indexed: 03/14/2025] Open
Abstract
BACKGROUND Emerging infectious diseases threaten human and animal health, with most pathogens originating from wildlife. Bats are natural hosts for many infectious agents. Previous studies have demonstrated that changes in some specific genes in bats may contribute to resistance to viral infections, but they have mostly overlooked the immune function of the bat gut microbiota. AIMS In this study, we used fecal transplants to transfer the gut microbiota from the Greater Horseshoe Bat (Rhinolophus ferrumequinum) into mice treated with antibiotics. The gut microbiota changes in mice were detected using 16S rRNA high-throughput sequencing technology. Flow cytometry was used to detect changes in associated immune cells in the spleen and mesenteric lymph nodes of the mice. RESULTS The results showed that the gut microbiota of mice showed characteristics of some bat gut microbiota. The Greater Horseshoe Bat's gut microbiota changed some immune cells' composition in the spleen and mesenteric lymph nodes of mice and also conferred a faster and higher proportion of natural killer cell activation. CONCLUSION This result provides new evidence for the regulatory immune function of bat gut microbiota and contributes to a deeper insight into the unique immune system of bats.
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Affiliation(s)
- Shan Luo
- College of Animal Science and Technology, Jilin Agricultural Science and Technology University, Jilin 132109, China
| | - Xinlei Huang
- College of Life Science, Jilin Agricultural University, Changchun 130117, China
| | - Siyu Chen
- College of Animal Science and Technology, Jilin Agricultural Science and Technology University, Jilin 132109, China
| | - Junyi Li
- College of Veterinary Medicine, Jilin Agricultural University, Changchun 130117, China
| | - Hui Wu
- College of Life Science, Jilin Agricultural University, Changchun 130117, China
| | - Yuhua He
- College of Animal Science and Technology, Jilin Agricultural Science and Technology University, Jilin 132109, China
| | - Lei Zhou
- College of Animal Science and Technology, Jilin Agricultural Science and Technology University, Jilin 132109, China
| | - Boyu Liu
- College of Animal Science and Technology, Jilin Agricultural Science and Technology University, Jilin 132109, China
| | - Jiang Feng
- College of Life Science, Jilin Agricultural University, Changchun 130117, China
- Jilin Provincial Key Laboratory of Animal Resource Conservation and Utilization, Northeast Normal University, Changchun 130024, China
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Hao W, Hu X, Chen Q, Qin B, Tian Z, Li Z, Hou P, Zhao R, Balci H, Cui S, Diao J. Duplex Unwinding Mechanism of Coronavirus MERS-CoV nsp13 Helicase. CHEMICAL & BIOMEDICAL IMAGING 2025; 3:111-122. [PMID: 40018651 PMCID: PMC11863148 DOI: 10.1021/cbmi.4c00077] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/04/2024] [Revised: 12/06/2024] [Accepted: 12/09/2024] [Indexed: 03/01/2025]
Abstract
The COVID-19 pandemic has underscored the importance of in-depth research into the proteins encoded by coronaviruses (CoV), particularly the highly conserved nonstructural CoV proteins (nsp). Among these, the nsp13 helicase of severe pathogenic MERS-CoV, SARS-CoV-2, and SARS-CoV is one of the most preserved CoV nsp. Utilizing single-molecule FRET, we discovered that MERS-CoV nsp13 unwinds DNA in distinct steps of about 9 bp when ATP is employed. If a different nucleotide is introduced, these steps diminish to 3-4 bp. Dwell-time analysis revealed 3-4 concealed steps within each unwinding process, which suggests the hydrolysis of 3-4 dTTP. Combining our observations with previous studies, we propose an unwinding model of CoV nsp13 helicase. This model suggests that the elongated and adaptable 1B-stalk of nsp13 may enable the 1B remnants to engage with the unwound single-stranded DNA, even as the helicase core domain has advanced over 3-4 bp, thereby inducing accumulated strain on the nsp13-DNA complex. Our findings provide a foundational framework for determining the unwinding mechanism of this unique helicase family.
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Affiliation(s)
- Wei Hao
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Xiao Hu
- Department
of Cancer Biology, University of Cincinnati
College of Medicine, Cincinnati, Ohio 45267, United States
| | - Qixin Chen
- Department
of Cancer Biology, University of Cincinnati
College of Medicine, Cincinnati, Ohio 45267, United States
| | - Bo Qin
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Zhiqi Tian
- Department
of Cancer Biology, University of Cincinnati
College of Medicine, Cincinnati, Ohio 45267, United States
| | - Ziheng Li
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Pengjiao Hou
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Rong Zhao
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Hamza Balci
- Department
of Physics, Kent State University, Kent, Ohio 44242, United States
| | - Sheng Cui
- NHC
Key Laboratory of Systems Biology of Pathogens, National Institute
of Pathogen Biology, Chinese Academy of
Medical Sciences and Peking Union Medical College, Beijing 100730, China
| | - Jiajie Diao
- Department
of Cancer Biology, University of Cincinnati
College of Medicine, Cincinnati, Ohio 45267, United States
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Neuman BW, Smart A, Gilmer O, Smyth RP, Vaas J, Böker N, Samborskiy DV, Bartenschlager R, Seitz S, Gorbalenya AE, Caliskan N, Lauber C. Giant RNA genomes: Roles of host, translation elongation, genome architecture, and proteome in nidoviruses. Proc Natl Acad Sci U S A 2025; 122:e2413675122. [PMID: 39928875 PMCID: PMC11848433 DOI: 10.1073/pnas.2413675122] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/25/2024] [Accepted: 01/09/2025] [Indexed: 02/12/2025] Open
Abstract
Positive-strand RNA viruses of the order Nidovirales have the largest known RNA genomes of vertebrate and invertebrate viruses with 36.7 and 41.1 kb, respectively. The acquisition of a proofreading exoribonuclease (ExoN) by an ancestral nidovirus enabled crossing of the 20 kb barrier. Other factors constraining genome size variations in nidoviruses remain poorly defined. We assemble 76 genome sequences of invertebrate nidoviruses from >500.000 published transcriptome experiments and triple the number of known nidoviruses with >36 kb genomes, including a 64 kb RNA genome. Many of the identified viral lineages acquired putative enzymatic and other protein domains linked to genome size, host phyla, or virus families. The inserted domains may regulate viral replication and virion formation, or modulate infection otherwise. We classify ExoN-encoding nidoviruses into seven groups and four subgroups, according to canonical and noncanonical modes of viral replicase expression by ribosomes and genomic organization (reModes). The most-represented group employing the canonical reMode comprises invertebrate and vertebrate nidoviruses, including coronaviruses. Six groups with noncanonical reModes include invertebrate nidoviruses with 31-to-64 kb genomes. Among them are viruses with segmented genomes and viruses utilizing dual ribosomal frameshifting that we validate experimentally. Moreover, largest polyprotein length and genome size in nidoviruses show reMode- and host phylum-dependent relationships. We hypothesize that the polyprotein length increase in nidoviruses may be limited by the host-inherent translation fidelity, ultimately setting a nidovirus genome size limit. Thus, expansion of ExoN-encoding RNA virus genomes, the vertebrate/invertebrate host division, the control of viral replicase expression, and translation fidelity are interconnected.
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Affiliation(s)
- Benjamin W. Neuman
- Department of Biology, Microbial Pathogenesis and Immunity, Texas A&M University, College Station, TX77840
| | - Alexandria Smart
- Helmholtz Institute for RNA-Based Infection Research, Helmholtz Centre for Infection Research, Würzburg97080, Germany
| | - Orian Gilmer
- Helmholtz Institute for RNA-Based Infection Research, Helmholtz Centre for Infection Research, Würzburg97080, Germany
| | - Redmond P. Smyth
- Helmholtz Institute for RNA-Based Infection Research, Helmholtz Centre for Infection Research, Würzburg97080, Germany
- Institut de Biologie Moléculaire et Cellulaire, Architecture et Réactivité de l’ARN, Université de Strasbourg, Strasbourg67084, France
| | - Josef Vaas
- Division of Virus-Associated Carcinogenesis (F170), German Cancer Research Center, Heidelberg 69120, Germany
- Medical Faculty Heidelberg, Department of Infectious Diseases, Molecular Virology, Heidelberg University, Center for Integrative Infectious Disease Research, Heidelberg69120, Germany
| | - Nicolai Böker
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover30625, Germany
- Cluster of Excellence 2155 RESIST, Hannover30625, Germany
| | - Dmitry V. Samborskiy
- Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow119899, Russia
| | - Ralf Bartenschlager
- Division of Virus-Associated Carcinogenesis (F170), German Cancer Research Center, Heidelberg 69120, Germany
- Medical Faculty Heidelberg, Department of Infectious Diseases, Molecular Virology, Heidelberg University, Center for Integrative Infectious Disease Research, Heidelberg69120, Germany
| | - Stefan Seitz
- Division of Virus-Associated Carcinogenesis (F170), German Cancer Research Center, Heidelberg 69120, Germany
- Medical Faculty Heidelberg, Department of Infectious Diseases, Molecular Virology, Heidelberg University, Center for Integrative Infectious Disease Research, Heidelberg69120, Germany
| | - Alexander E. Gorbalenya
- Leiden University Center of Infectious Diseases, Leiden University Medical Center, Leiden2333 ZA, The Netherlands
| | - Neva Caliskan
- Helmholtz Institute for RNA-Based Infection Research, Helmholtz Centre for Infection Research, Würzburg97080, Germany
- Department of Biochemistry III, University of Regensburg, Regensburg93053, Germany
| | - Chris Lauber
- Institute for Experimental Virology, TWINCORE Centre for Experimental and Clinical Infection Research, a Joint Venture between the Hannover Medical School and the Helmholtz Centre for Infection Research, Hannover30625, Germany
- Cluster of Excellence 2155 RESIST, Hannover30625, Germany
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Arias-Arias JL, Monturiol-Gross L, Corrales-Aguilar E. A Live-Cell Imaging-Based Fluorescent SARS-CoV-2 Neutralization Assay by Antibody-Mediated Blockage of Receptor Binding Domain-ACE2 Interaction. BIOTECH 2025; 14:10. [PMID: 39982277 PMCID: PMC11843899 DOI: 10.3390/biotech14010010] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/12/2024] [Revised: 01/15/2025] [Accepted: 01/17/2025] [Indexed: 02/22/2025] Open
Abstract
Neutralization assays have become an important tool since the beginning of the COVID-19 pandemic for testing vaccine responses and therapeutic antibodies as well as for monitoring humoral immunity to SARS-CoV-2 in epidemiological studies. The spike glycoprotein (S) present on the viral surface contains a receptor binding domain (RBD) that recognizes the angiotensin-converting enzyme 2 receptor (ACE2) in host cells, allowing virus entry. The gold standard for determining SARS-CoV-2 neutralizing antibodies is the plaque reduction neutralization test (PRNT), which relies on live-virus replication performed exclusively in biosafety level 3 (BSL-3) laboratories. Here, we report the development of a surrogate live-cell imaging-based fluorescent SARS-CoV-2 neutralization assay, applicable to BSL-1 or BSL-2 laboratories, by antibody-mediated blockage of the interaction between recombinant RBD with overexpressed ACE2 receptor in a genetically modified HEK 293T stable cell line. Our approach was able to detect neutralizing antibodies both in COVID-19-positive human serum samples and polyclonal equine formulations against SARS-CoV-2. This new cell-based surrogate neutralization assay represents a virus-free fluorescence imaging alternative to the reported approaches, which can be used to detect antibody-neutralizing capabilities toward SARS-CoV-2. This assay could also be extrapolated in the future to other established and emergent viral agents.
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Affiliation(s)
- Jorge L. Arias-Arias
- Centro de Investigación en Enfermedades Tropicales (CIET), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica;
- Dulbecco Lab Studio, Residencial Lisboa 2G, Alajuela 20102, Costa Rica
| | - Laura Monturiol-Gross
- Instituto Clodomiro Picado (ICP), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica;
| | - Eugenia Corrales-Aguilar
- Centro de Investigación en Enfermedades Tropicales (CIET), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica;
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Flury P, Krüger N, Sylvester K, Breidenbach J, Al Hamwi G, Qiao J, Chen Y, Rocha C, Serafim MSM, Barbosa da Silva E, Pöhlmann S, Poso A, Kronenberger T, Rox K, O'Donoghue AJ, Yang S, Sträter N, Gütschow M, Laufer SA, Müller CE, Pillaiyar T. Design, Synthesis, and Unprecedented Interactions of Covalent Dipeptide-Based Inhibitors of SARS-CoV-2 Main Protease and Its Variants Displaying Potent Antiviral Activity. J Med Chem 2025; 68:3626-3652. [PMID: 39813204 DOI: 10.1021/acs.jmedchem.4c02254] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2025]
Abstract
The main protease (Mpro) of SARS-CoV-2 is a key drug target for the development of antiviral therapeutics. Here, we designed and synthesized a series of small-molecule peptidomimetics with various cysteine-reactive electrophiles. Several compounds were identified as potent SARS-CoV-2 Mpro inhibitors, including compounds 8n (IC50 = 0.0752 μM), 8p (IC50 = 0.0887 μM), 8r (IC50 = 0.0199 μM), 10a (IC50 = 0.0376 μM), 10c (IC50 = 0.0177 μM), and 10f (IC50 = 0.0130 μM). Most of them additionally inhibited cathepsin L and were also active against SARS-CoV-1 and MERS-CoV Mpro. In Calu-3 cells, several inhibitors, including 8r, 10a, and 10c, displayed high antiviral activity in the nanomolar range without showing cellular toxicity. The cocrystal structure of SARS-CoV-2 Mpro in complex with 8p revealed covalent binding to the enzyme's catalytic residue Cys145 and showed specific, unprecedented interactions within the substrate binding pocket. Compounds 10c and especially 8n were effective against a panel of naturally occurring nirmatrelvir-resistant mutants, particularly E166V, and showed metabolic stability and additional favorable pharmacokinetic properties, making it a suitable candidate for further preclinical development.
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Affiliation(s)
- Philipp Flury
- Institute of Pharmacy, Pharmaceutical/Medicinal Chemistry and Tübingen Center for Academic Drug Discovery, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
| | - Nadine Krüger
- Platform Infection Models, German Primate Center, Leibniz Institute for Primate Research Göttingen, Kellnerweg 4, 37077 Göttingen, Germany
| | - Katharina Sylvester
- PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121 Bonn, Germany
| | - Julian Breidenbach
- PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121 Bonn, Germany
| | - Ghazl Al Hamwi
- PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121 Bonn, Germany
| | - Jingxin Qiao
- Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China
| | - Yan Chen
- Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China
| | - Cheila Rocha
- Infection Biology Unit, German Primate Center - Leibniz Institute for Primate Research, Kellnerweg 4, Göttingen 37077, Germany
| | - Mateus Sá Magalhães Serafim
- Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte 31270-901, Minas Gerais, Brazil
- Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093-0657, United States
| | - Elany Barbosa da Silva
- Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093-0657, United States
| | - Stefan Pöhlmann
- Infection Biology Unit, German Primate Center - Leibniz Institute for Primate Research, Kellnerweg 4, Göttingen 37077, Germany
- Faculty of Biology and Psychology, Georg-August University Göttingen, Göttingen, 37073, Germany
| | - Antti Poso
- Institute of Pharmacy, Pharmaceutical/Medicinal Chemistry and Tübingen Center for Academic Drug Discovery, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
- School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio 70211, Finland
| | - Thales Kronenberger
- Institute of Pharmacy, Pharmaceutical/Medicinal Chemistry and Tübingen Center for Academic Drug Discovery, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
- School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Kuopio 70211, Finland
- German Center for Infection Research (DZIF), Partner Site Tübingen, Elfriede-Aulhorn-Str. 6, Tübingen 72076, Germany
| | - Katharina Rox
- Department of Chemical Biology, Helmholtz Centre for Infection Research (HZI), Braunschweig 38124, Germany
- German Center for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Braunschweig 38124, Germany
| | - Anthony J O'Donoghue
- Center for Discovery and Innovation in Parasitic Diseases, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093-0657, United States
| | - Shengyong Yang
- Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China
| | - Norbert Sträter
- Center for Biotechnology and Biomedicine, Leipzig University, Deutscher Platz 5, Leipzig 04103, Germany
| | - Michael Gütschow
- PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121 Bonn, Germany
| | - Stefan A Laufer
- Institute of Pharmacy, Pharmaceutical/Medicinal Chemistry and Tübingen Center for Academic Drug Discovery, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
- Cluster of Excellence "Image Guided and Functionally Instructed Tumor Therapies" (iFIT), Eberhard Karls University of Tuebingen, Tuebingen 72076, Germany
| | - Christa E Müller
- PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical & Medicinal Chemistry, University of Bonn, An der Immenburg 4, 53121 Bonn, Germany
| | - Thanigaimalai Pillaiyar
- Institute of Pharmacy, Pharmaceutical/Medicinal Chemistry and Tübingen Center for Academic Drug Discovery, Eberhard Karls University Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
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Kim J, Kachko A, Selvaraj P, Rotstein D, Stauft CB, Rajasagi N, Zhao Y, Wang T, Major M. Combined immunization with SARS-CoV-2 spike and SARS-CoV nucleocapsid protects K18-hACE2 mice but increases lung pathology. NPJ Vaccines 2025; 10:30. [PMID: 39948345 PMCID: PMC11825953 DOI: 10.1038/s41541-025-01085-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2024] [Accepted: 01/25/2025] [Indexed: 02/16/2025] Open
Abstract
Vaccines against SARS-CoV-2 have targeted the spike protein and have been successful at preventing disease. However, with the emergence of variants, spike-specific vaccines become less effective. The nucleocapsid protein is relatively conserved among variants of SARS-CoV-2 and is a candidate for addition to spike in next generation vaccines for the induction of T cell protection. Previous studies on SARS-CoV have suggested that the induction of an immune response to nucleocapsid could result in enhanced disease. Using the K18-hACE2 mouse model we investigated immunization with a variant nucleocapsid, from SARS CoV (N1) alone or in combination with spike from SARS-CoV-2 and compared this to nucleocapsid from SARS-CoV-2 (N2). The spike-nucleocapsid-based vaccines conferred protection against SARS-CoV-2 in lungs and brain and decreased lung pathology compared to control mice. However, higher T and B cell immune responses were observed in N1-immunized mice prior to challenge, whether delivered alone or with spike, and immunization with N1 resulted in increased lung pathology compared to immunization with spike or N2. These findings suggest that spike-nucleocapsid-based vaccines are safe and effective, even with variant nucleocapsid sequences, but that viral control in this mouse model may be associated with higher lung pathology, compared to spike immunization alone, due to the immunogenic qualities of the nucleocapsid antigen.
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Affiliation(s)
- Jaekwan Kim
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Alla Kachko
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Prabhuanand Selvaraj
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - David Rotstein
- Division of Food Compliance, Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD, USA
| | - Charles Brandon Stauft
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Naveen Rajasagi
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Yangqing Zhao
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Tony Wang
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA
| | - Marian Major
- Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, USA.
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