• H pylori
• cell therapy
• chronic atrophic gastritis
• inflammation
• placenta-derived mesenchymal stem cell
• rejuvenation

•

Haplotype blocks methods plays a complementary role to the single-SNP approaches.

The human genome, which includes thousands of genes, represents a big data challenge. Rheumatoid arthritis (RA) is a complex autoimmune disease with a genetic basis. Many single-nucleotide polymorphism (SNP) association methods partition a genome into haplotype blocks. The aim of this genome wide association study (GWAS) was to select the most appropriate haplotype block partitioning method for the North American Rheumatoid Arthritis Consortium (NARAC) dataset. The methods used for the NARAC dataset were the individual SNP approach and the following haplotype block methods: the four-gamete test (FGT), confidence interval test (CIT), and solid spine of linkage disequilibrium (SSLD). The measured parameters that reflect the strength of the association between the biomarker and RA were the P-value after Bonferroni correction and other parameters used to compare the output of each haplotype block method. This work presents a comparison among the individual SNP approach and the three haplotype block methods to select the method that can detect all the significant SNPs when applied alone. The GWAS results from the NARAC dataset obtained with the different methods are presented. The individual SNP, CIT, FGT, and SSLD methods detected 541, 1516, 1551, and 1831 RA-associated SNPs respectively, and the individual SNP, FGT, CIT, and SSLD methods detected 65, 156, 159, and 450 significant SNPs respectively, that were not detected by the other methods. Three hundred eighty-three SNPs were discovered by the haplotype block methods and the individual SNP approach, while 1021 SNPs were discovered by all three haplotype block methods. The 383 SNPs detected by all the methods are promising candidates for studying RA susceptibility. A hybrid technique involving all four methods should be applied to detect the significant SNPs associated with RA in the NARAC dataset, but the SSLD method may be preferred because of its advantages when only one method was used.

• Confidence interval test
• Four-gamete test
• Genome-wide association study
• NARAC
• Rheumatoid arthritis
• Solid spine of linkage disequilibrium

• gastric cancer
• immunohistochemistry
• musashi-1
• prognosis
• tissue microarray
• western blotting

Foamy macrophages (FM)s harbor lipid bodies that not only assist mycobacterial persistence within the granulomas but also are sites for intracellular signaling and inflammatory mediators which are essential for mycobacterial pathogenesis. However, molecular mechanisms that regulate intracellular lipid accumulation in FMs during mycobacterial infection are not clear. Here, we report for the first time that jumonji domain containing protein (JMJD)3, a demethylase of the repressive H3K27me3 mark, orchestrates the expression of M. tuberculosis H37Rv-, MDR-JAL2287-, H37Ra- and M. bovis BCG-induced genes essential for FM generation in a TLR2-dependent manner. Further, NOTCH1-responsive RNA-binding protein MUSASHI (MSI), targets a transcriptional repressor of JMJD3, Msx2-interacting nuclear target protein, to positively regulate infection-induced JMJD3 expression, FM generation and M2 phenotype. Investigations in in vivo murine models further substantiated these observations. Together, our study has attributed novel roles for JMJD3 and its regulators during mycobacterial infection that assist FM generation and fine-tune associated host immunity.

Foamy macrophages (FMs) not only provide a suitable survival niche for the mycobacteria in the granuloma but also are reservoirs for several inflammatory mediators that regulate mycobacterial pathogenesis. Hence, understanding the mechanisms that regulate infection-induced FM generation assumes importance. In this investigation, we present empirical evidence to support the role of host epigenetic mechanisms in generating FMs and thus facilitating mycobacterial persistence in vivo. We show that the signaling pathways that mediate mycobacteria-induced expression of JMJD3, a demethylase of the facultative repression mark, regulate the genes assisting in FM generation. Importantly, the identified pathway could largely contribute to the evasive responses during mycobacterial infection and suppression of such pathways during infection could confer stronger immunity. Together, these regulators could be potential candidates for host-directed therapies against mycobacterial infection.

• Animals
• Chromatin Immunoprecipitation
• Disease Models, Animal
• Fluorescent Antibody Technique
• Gene Expression Regulation, Bacterial/immunology
• Granuloma/immunology
• Granuloma/microbiology
• Immunoblotting
• Immunohistochemistry
• Immunoprecipitation
• Jumonji Domain-Containing Histone Demethylases/immunology
• Jumonji Domain-Containing Histone Demethylases/metabolism
• Macrophages/immunology
• Macrophages/metabolism
• Macrophages/microbiology
• Mice
• Mycobacterium Infections/immunology
• Mycobacterium Infections/metabolism
• Mycobacterium tuberculosis/immunology
• Mycobacterium tuberculosis/metabolism
• Nerve Tissue Proteins/immunology
• Nerve Tissue Proteins/metabolism
• RNA-Binding Proteins/immunology
• RNA-Binding Proteins/metabolism
• Real-Time Polymerase Chain Reaction
• Transfection
• Tuberculosis/immunology
• Tuberculosis/metabolism

• Wellcome Trust
• 500034-Z-09-Z DBT-Wellcome Trust India Alliance
• Department of Biotechnology, Ministry of Science and Technology
• IFCPAR/CEFIPRA
• Wellcome-DBT India Alliance

AIM: To investigate expression of stem cell marker Musashi-1 (Msi-1) in relationship to tumorigenesis and progression of intestinal-type gastric cancer (GC).

METHODS: Endoscopic biopsy specimens and surgical specimens were obtained, including 54 cases of intestinal-type GC, 41 high-grade intraepithelial neoplasia, 57 low-grade intraepithelial neoplasia, 31 intestinal metaplasia, and 36 normal gastric mucosa. Specimens were fixed in 10% paraformaldehyde, conventionally dehydrated, embedded in paraffin, and sliced in 4-μm-thick serial sections. Two-step immunohistochemical staining was used to detect Msi-1 and proliferating cell nuclear antigen (PCNA) expression. Correlation analysis was conducted between Msi-1 and PCNA expression. The relationship between Msi-1 expression and clinicopathological parameters of GC was analyzed statistically.

RESULTS: There were significant differences in Msi-1 and PCNA expression in different pathological tissues (χ² = 15.37, P < 0.01; χ² = 115.36, P < 0.01). Msi-1 and PCNA-positive cells were restricted to the isthmus of normal gastric glands. Expression levels of Msi-1 and PCNA in intestinal metaplasia were significantly higher than in normal mucosa (U = 392.0, P < 0.05; U = 40.50, P < 0.01), whereas there was no significant difference compared to low or high-grade intraepithelial neoplasia. Msi-1 and PCNA expression in intestinal-type GC was higher than in high-grade intraepithelial neoplasia (U = 798.0, P < 0.05; U = 688.0, P < 0.01). There was a significantly positive correlation between Msi-1 and PCNA expression (r_s = 0.20, P < 0.01). Msi-1 expression in GC tissues was correlated with their lymph node metastasis and tumor node metastasis stage (χ² = 12.62, P < 0.01; χ² = 11.24, P < 0.05), but not with depth of invasion and the presence of distant metastasis.

CONCLUSION: Msi-1-positive cells may play a key role in the early events of gastric carcinogenesis and may be involved in invasion and metastasis of GC.

• Musashi-1
• Stem cells
• Gastric cancer
• Precancerous lesions
• Immunohistochemistry.

• Adult
• Aged
• Biomarkers, Tumor/analysis
• Biopsy
• Carcinoma in Situ/chemistry
• Carcinoma in Situ/pathology
• Cell Proliferation
• Cell Transformation, Neoplastic/chemistry
• Cell Transformation, Neoplastic/pathology
• Chi-Square Distribution
• Disease Progression
• Female
• Gastroscopy
• Humans
• Immunohistochemistry
• Lymphatic Metastasis
• Male
• Metaplasia
• Middle Aged
• Neoplasm Invasiveness
• Neoplasm Staging
• Neoplastic Stem Cells/chemistry
• Neoplastic Stem Cells/pathology
• Nerve Tissue Proteins/analysis
• Precancerous Conditions/chemistry
• Precancerous Conditions/pathology
• Prognosis
• Proliferating Cell Nuclear Antigen/analysis
• RNA-Binding Proteins/analysis
• Stomach Neoplasms/chemistry
• Stomach Neoplasms/pathology

AIM: To observe the effect of CagA⁺Helicobacter pylori (H. pylori) on the expression of HIF-2α and ABCG2 in human gastric cancer cell line SGC7901 under normoxia and hypoxia conditions to investigate whether H. pylori infection and the tumor microenvironment have a synergistic effect in the initiation and development of gastric cancer.

METHODS: Gastric mucosal biopsy specimens collected by endoscopy were cultured under microaerophilic conditions and H. pylori isolates were identified. CagA⁺H. pylori strains were confirmed by PCR. Gastric cancer cell line SGC7901 was co-cultured with a CagA⁺H. pylori strain for 48 h under either normoxia or hypoxia condition (cells were divided into a normoxia control group, a hypoxia control group, a normoxia plus CagA⁺H. pylori group, and a hypoxia plus CagA⁺H. pylori group). Immunocytochemistry was used to detect the expression of HIF-2α and ABCG2 proteins, and RT-PCR was used to detect the expression of ABCG2 mRNA.

RESULTS: Immunocytochemistry results showed that HIF-2α and ABCG2 proteins were expressed at low levels under normoxia, while both hypoxia and CagA⁺H. pylori could significantly induce the expression of HIF-2α and ABCG2 proteins compared to the normoxia control group (all P < 0.01). Compared to the hypoxia control group and normoxia plus CagA⁺H. pylori group, the expression of HIF-2α and ABCG2 proteins was further elevated in the hypoxia plus CagA⁺H. pylori group (all P < 0.01). There was a positive correlation between the expression of HIF-2α and that of ABCG2 (r = 0.976, P < 0.05). Similar results were obtained for ABCG2 mRNA expression by RT-PCR.

CONCLUSION: CagA⁺H. pylori can stimulate the expression of HIF-2α and ABCG2 in gastric cancer cells under normoxia condition, and their expression can be further up-regulated under hypoxia condition. CagA⁺H. pylori and hypoxia have a synergistic effect on the expression of HIF-2α and ABCG2, suggesting that CagA⁺H. pylori and hypoxia may play an important role in inducing gastric cancer cell de-differentiation and chemotherapy resistance.

• Gastric cancer cells
• Helicobacter pylori
• Cytotoxin-associated protein A
• Hypoxia
• ABCG2
• Hypoxia inducible factor-2alpha

• gastric cancer
• growth factor/cytokine
• tumor microenvironment
• helicobacter pylori
• inflammation
• nf-kb pathway