Up-regulation of extracellular-matrix and inflammation related genes in oral squamous cell carcinoma.

OBJECTIVE: Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy with late-presentation, site-specific heterogeneity, and high-propensity for recurrence/metastasis that has shown rise in mortality. Lately, research emphasize on dynamic interactions between tumor-cells and extracellular-matrix components within tumor-microenvironment that promote tissue integrity loss and carcinogenesis. Therefore, OSCC clinical-management is still challenging. DESIGN: Present study validated clinical utility of a 13 gene-panel in two chief sub-sites of OSCC: Buccal mucosa squamous cell carcinoma (BMSCC) (N = 50) and Tongue squamous cell carcinoma (TSCC) (N = 52) using qRT-PCR. Principal component analysis and binary logistic regression analysis were applied to acquire definite multi gene models. Protein expression analysis was employed using the Human Protein Atlas, UALCAN and TIMER 2.0 databases to explore potential correlation between immune cells and gene-panels. RESULTS: Significant up-regulation of CXCL8, CXCL10, FN1, GBP1, IFIT3, ISG15, MMP1, MMP3, MMP10, PLAU, SERPINE1 and SPP1 except OASL was observed in OSCC tissue in comparison of absolute normal controls. Although, this gene-panel could potentially discriminate OSCC tissues from absolute normal controls as solitarily diagnostic and/or predictive biomarkers, models generated also showed substantial discriminating efficacy. Eight-genes were found to be significantly associated with poor-prognosis on clinico-pathological association. Protein-expression confirmed overexpression of gene-panel and added advantage of being secretory-protein. Importantly, up-regulated genes in our study showed significant relation with immune-cells infiltration suggesting their contribution in immune-escape. CONCLUSION: Thus, we propose that the 13 gene-panel could pave the way to effective and personalized clinical-management of OSCC in terms of diagnostic and prognostic measures and thereby as therapeutic targets.

Association of FTO gene variant rs9939609 with polycystic ovary syndrome from Gujarat, India.

BACKGROUND: Polycystic ovary syndrome is a multifactorial endocrine disorder impacting women of reproductive age. Variations within the FTO gene have been linked to both obesity and type 2 diabetes mellitus. Given that PCOS is frequently associated with obesity and compromised glucose tolerance, we investigated the prevalence of the rs9939609 variant within the FTO gene among women diagnosed with PCOS and a control group. Our aim is to uncover potential correlations between this genetic variant, metabolic attributes, and endocrine markers within the Gujarat province of India. METHOD: We enrolled a total of 114 participants, (62 individuals diagnosed with PCOS and 52 healthy controls). DNA extraction from venous blood was conducted for all participants. The rs9939609 polymorphism was investigated through tetra-primer amplification refractory mutation system-polymerase chain reaction. Furthermore, we performed biochemical assessments to quantify levels of estradiol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), total testosterone, prolactin (PRL), and Dehydroepiandrosterone sulfate (DHEAS). Statistical analyses were carried out utilizing SPSS version 21 (IBM, USA). RESULTS: The present study did not reveal any noteworthy association between cases and controls. The frequencies of genotypes and alleles within the cohorts displayed no statistically significant differences (p = 0.25, p = 0.68, and p = 0.78, respectively). The dominant model indicated a modest risk (OR:1.13, 95%CI: 0.55 to 2.38) toward PCOS development. There was a noticeable statistical difference observed in the levels of total testosterone, DHEAS, and BMI between the case and control groups (p < 0.002, p < 0.0002, p < 0.0008). However, no variations in clinical variables were observed among genotypes within the PCOS group. CONCLUSION: This is the first study to investigate the association of FTO gene polymorphism and PCOS in Gujarati population. Our study findings indicate that the FTO gene variant is not directly linked to the onset of PCOS. However, it appears to exert an influence on metabolic factors such as obesity and insulin resistance. Notably, our results suggest that insulin resistance is more frequently observed among PCOS patients who are obese, as compared to those with non-obese PCOS patients.

An Updated Mutation Spectrum of the γ-Secretase Complex: Novel NCSTN Gene Mutation in an Indian Family with Hidradenitis Suppurativa and Acne Conglobata.

Background: Hidradenitis suppurativa (HS) is a complex, chronic inflammatory skin disorder whose pathophysiology is poorly understood. Genetic studies have shown that HS is predisposed by mutations in the γ-secretase gene, but only a proportion of familial and partial sporadic cases have been shown to possess such mutations. HS has high genetic heterogeneity and is thought to be triggered by a combination of genetics and environmental factors. Aims: The study aimed to investigate the genetic causes of HS in a large cohort of patients and to update the mutation spectrum of γ-secretase complex genes. Methods: We conducted mutational screening of 95 sporadic HS cases and one large family with both HS and acne conglobata (AC) to identify mutations in the coding and splice junction region of γ-secretase complex genes (nicastrin (NCSTN), presenilin 1 (PSEN1), presenilin enhancer 2 (PSENEN), and aph-1 homolog B, gamma-secretase subunit (APH1B)). Results: Our study identified a nucleotide substitution of 1876C>T in the NCSTN gene, which caused a stop codon (p.Arg626X) in the affected members of a large family with HS and AC. No pathogenic variants were detected in 95 sporadic cases of HS, indicating there is possible genetic heterogeneity. Conclusion: We report a new family with a nonsense mutation in the NCSTN gene that supports the role of the γ-secretase complex genes in HS with AC. The updated γ-secretase mutation spectrum for HS now includes 78 mutations.

Uncovering the Interaction Interface Between Harpin (Hpa1) and Rice Aquaporin (OsPIP1;3) Through Protein-Protein Docking: An In Silico Approach.

Hpa1 (a type of harpin) is involved in T3SS (Type III Secretion System) assembly in the infection mechanism by Xanthomonas Oryzae pv. oryzae (Xoo). Hpa1 interacts with the plasma membrane components of plants thereby assisting effector proteins toward the cytoplasm, wherein effectors execute their pathological functions. Independently, harpins also induce hypersensitive response and systemic acquired resistance in plants. However, lack of knowledge regarding the plant-harpin interaction mechanism constrains the pathway of its agricultural application. Although an in vitro study proved that Hpa1 protein can interact with OsPIP1;3, a rice aquaporin, the structural basis of the interaction is yet to be discovered. The presented work is the first of its kind where an in silico approach is used for the PPI (protein-protein interaction) of harpin protein. The study discovered participation of Hpa1 N-terminal amino acids at the interface. Besides, MD simulation studies were performed to assess the stability. RMSD values were 0.35 ± 0.049, 0.73 ± 0.11, and 0.50 ± 0.065 nm for OsPIP1;3, Hpa1, and Hpa1-OsPIP1;3 complex, respectively. Additionally, Residue-wise fluctuations have also been studied post-MDS. Taken together, these findings not only give a solid foundation for a deeper knowledge of various interacting target molecules with Harpin protein orthologs but also bring a new avenue for the structural-functional relationship study of harpin proteins.

Curcumin mediated dendritic cell maturation by modulating cancer associated fibroblasts-derived exosomal miRNA-146a.

BACKGROUND: Though cancer associated fibroblasts (CAFs), being a main component of tumor microenvironment (TME), are known to modulate immune response through secretion of various growth hormones, exosomes carrying miRNAs and cytokines; their effect on dendritic cells (DCs) are yet to be elucidated. Thus, aim of this study was to assess the effect of miRNAs and cytokines released by lung-CAFs and to evaluate immunomodulatory potential of curcumin on DC maturation through modulating their TME. MATERIAL AND METHODS: To check the effect of CAFs derived exosomes on DC maturation, we cultured imDCs in the presence of CAFs derived conditioned media (CAFs-CM) and characterized by the presence of maturation markers CD80, CD83, CD86 and CTLA4 using qRT-PCR. Additionally, expression of miR-221, miR-222, miR-155, miR-142-3p and miR-146a was assessed to evaluate the role of epigenetic regulators on DC maturation. Likewise, cytokine profiling of CAFs-CM as well as CAFs-CM treated with curcumin was also conducted using ELISA. RESULTS: Results revealed the generation of regulatory DCs which were characterized by decreased expression of maturation markers in the presence of CAFs-CM. In addition, such DCs showed higher expression of epigenetic regulator miR-146a which was positively correlated with increased expression of anti-inflammatory cytokines like IL-6, IL-10, TGF-ß and decreased expression of TNF-α (pro-inflammatory). Moreover, curcumin had the potential to convert regulatory DCs generated by CAFs into mDCs, which were characterized by high expression of co-stimulatory molecules, low expression of CTLA4, lower levels of immune suppressive cytokines production and lower levels of miR-146a. CONCLUSION: Collectively, these findings provide insight into understanding the immunomodulatory role of curcumin in targeting CAFs and modulating TME, thus enhancing antitumor immune response in DC based therapy.

An integrative analysis to enumerate candidate genes for clinical use in oral cancer.

Background: Oral cancer (OC) is the most pernicious sub-site of head and neck tumours with poor prognostic value that is largely ascribed to the lack of ideal biomarkers and therapeutic targets. This fact highlights an urgent need to identify biomarkers that can further aid in OC management. Aim: The aim of this study was to identify a gene panel with a maximum clinical utility for OC. Materials and Methods: Eight eligible datasets were downloaded from the Gene Expression Omnibus Database, containing 320OC samples and 173 normal samples. The data were processed by GeneSpring software to reveal differentially expressed genes between OC tissues and normal tissues in eight individual experiments. Functional enrichment and network analysis were performed using PANTHER and STRING databases for concordant genes (fold change >10; P ≤ 0.05). The selected genes were cross-validated in the cancer genome atlas (TCGA), Oncomine, and KaplanMeier (KM) plotter databases. Results: Totally, 65 concordant genes were identified, including 37 up-regulated genes and 28 down-regulated genes. A 13-gene panel CXCL8, CXCL10, FN1, GBP1, IFIT3, ISG15, MMP1, MMP3, MMP10, OASL, SERPINE1, SPP1, and PLAU was elected from the lists of functionally enriched genes, hub genes, and genes that showed high alterations for mutation, copy number variation, and mRNA expression status in 'Head and Neck Squamous Cell Carcinoma patients (n = 279; TCGA, Nature 2015)'. Further, validation in Oncomine database demonstrated significant over-expression of all elected genes in OC patients across multiple datasets. In addition, out of 13, six genes (CXCL8, CXCL10, FN1, PLAU, SERPINE1, and SPP1) showed significant association with the prognosis of Head and Neck cancer patients (n = 500) in the KM plotter database. Conclusions: Using an integrative analysis, our study investigated and validated a 13-gene panel for OC which can be used to improve current diagnostic, prognostic, and treatment approaches.

Assessing the Protective Effect of Moringa oleifera Extract against Bone Metastasis: An In Vitro Simulated Digestion Approach.

Moringa oleifera possesses numerous advantageous effects like anti-microbial, antioxidant, and anti-inflammatory, leaves contain a high multiplicity of the bioactive compound; however, little is identified about its bioaccessibility. The objective of this study was to assess the bioefficacy, bioaccessible and anticancer activity of Moringa oleifera in a PC3 cell line before and after simulated in vitro digestion. Digested and non-digested extracts were prepared and evaluated for total polyphenols, flavonoids, and total antioxidant capacity by spectrophotometric analysis and LCMS analysis. Cell viability, apoptosis, colony formation, cell cycle, Glutathione level, and gene expression study were tested with Moringa oleifera (MO) and digested Moringa oleifera (DMO). Results revealed that total polyphenols, total flavonoids, and TAC were significantly (P < 0.05) reduced after in vitro digestion. Furthermore, biological activity against the PC3 cell line showed that DMO extracts significant cytotoxic and reduced cell vitality compared to the MO. In addition, DMO extract had a noteworthy effect in apoptosis and inhibiting the colony formation ability; while cell cycle was blocked in S phase by both extracts but significant effect showed in DMO. These studies have increased understanding of the influence of in vitro simulation digestion on the biological activity effect of M. oleifera against prostate cancer bone metastasis.Supplemental data for this article is available online at https://doi.org/10.1080/01635581.2021.1933099 .

The role of polymorphism in various potential genes on polycystic ovary syndrome susceptibility and pathogenesis.

Polycystic ovary syndrome (PCOS) is the most common endocrinopathies affecting the early reproductive age in women, whose pathophysiology perplexes many researchers till today. This syndrome is classically categorized by hyperandrogenism and/or hyperandrogenemia, menstrual and ovulatory dysfunction, bulky multi follicular ovaries on Ultrasonography (USG), and metabolic abnormalities such as hyperinsulinemia, dyslipidemia, obesity. The etiopathogenesis of PCOS is not fully elucidated, but it seems that the hypothalamus-pituitary-ovarian axis, ovarian, and/or adrenal androgen secretion may contribute to developing the syndrome. Infertility and poor reproductive health in women's lives are highly associated with elevated levels of androgens. Studies with ovarian theca cells taken from PCOS women have demonstrated increased androgen production due to augmented ovarian steroidogenesis attributed to mainly altered expression of critical enzymes (Cytochrome P450 enzymes: CYP17, CYP21, CYP19, CYP11A) in the steroid hormone biosynthesis pathway. Despite the heterogeneity of PCOS, candidate gene studies are the widely used technique to delineate the genetic variants and analyze for the correlation of androgen biosynthesis pathway and those affecting the secretion or action of insulin with PCOS etiology. Linkage and association studies have predicted the relationship between genetic variants and PCOS risk among families or populations. Several genes have been proposed as playing a role in the etiopathogenesis of PCOS, and the presence of mutations and/or polymorphisms has been discovered, which suggests that PCOS has a vital heritable component. The following review summarizes the influence of polymorphisms in crucial genes of the steroidogenesis pathway leading to intraovarian hyperandrogenism which can result in PCOS.

A predictive biomarker panel for bone metastases: Liquid biopsy approach.

Bone metastases is one of the common metastatic site and leading cause of cancer-related mortality in progressive cancer patients. The purpose of the present study is to establish a liquid biopsy based multi-gene classifier and associated signalling pathways for early diagnosis of bone metastases. We used publically available microarray datasets and analysed them in a platform/chip-specific manner using GeneSpring software. Analyses of gene expression datasets identified 15 consistently over-expressed genes with statistical significance. Further, expression profile of same set of 15 genes were compared in breast and lung cancer exosome derived mRNA with (n = 10) and without (n = 10) bone metastases against healthy controls. ROC curve analysis performed individually for all the 15 genes shortlisted the 5 most relevant genes with significant sensitivity and specificity in both cancers. This liquid biopsy-based bone metastases predictor using multi-gene panel is a unique approach with potential clinical applications for effective management of aggressive cancers.

Development of probiotic yogurt: effect of strain combination on nutritional, rheological, organoleptic and probiotic properties.

Seven combinations of yogurt; C1 [yogurt starter culture (YSC)], T1, [YSC + Lactobacillus acidophilus (LA)], T2 [YSC + Bifidobacterium bifidum (BB)], T3 [YSC + Lactobacillus plantarum (LP)], T4 [YSC + Lactobacillus casei (LC)], T5 [YSC + LA + BB] and T6 [YSC + LP + LC] were developed. Nutritional [proximate and minerals], rheological [total soluble solids (TSS), pH, titratable acidity (TA), water holding capacity, synersis, viscosity] organoleptic and probiotic properties [viability, acid tolerance, bile salt tolerance] were assessed with standard methods. Nutritional composition differed significantly among samples except for the iron and zinc (P < 0.05). Yogurt containing LP as single or in combination with LC resulted in significantly higher ash, protein, calcium and phosphorous level. Probiotic combination also significantly affected the rheological properties of yogurts (P < 0.05). Yogurt with LP and LC as single or in combination lead to significantly higher TSS and viscosity while significantly low syneresis, whereas yogurt with LA as single or in combination resulted in low pH and high TA (P < 0.05). Interestingly, combination of LA and BB increased TSS, reduced pH and syneresis as compare to these bacteria as single probiotic source. Panel experts found yogurt with LP more flavourful. Combination of multi-strain and multi-species probiotic resulted in improved texture but we found no significant difference in overall acceptability. Combination of probiotic strains also resulted in better probiotic potential with multi-species combination found to be even more effective. BB seemed more stable than three other probiotic strains. The present study can be helpful to dairy industry in developing new probiotic products and may provide a rational for selecting a combination of probiotic strains.