Associations of vitamin D receptor encoding gene variants with premenopausal breast cancer risk.

OBJECTIVE: This study evaluates whether vitamin D receptor (VDR) gene variations are associated with premenopausal breast cancer in Pakistani cohorts. METHODS: Genomic DNA was isolated from 228 breast cancer patients and 500 non-cancer controls. Six polymorphic variants (rs11568820, rs4516035, rs2228570, rs1544410, rs7975232, rs731236) of the VDR gene were genotyped using PCR-RFLP analysis. All statistical analysis was carried out on IBM-SPSS 23 at p-value <.05. Chi-square test and odds ratios (ORs) along with 95% confidence interval (CIs) were applied to evaluate the relationship between VDR gene polymorphisms and breast cancer. RESULTS: Results showed that the A/A genotype of EcoRV (OR = 2.125, 95% CI = 1.024 to 4.412) and the A/a genotype of Apa1 (OR = 6.094, 95%CI = 4.111 to 9.033) gene polymorphism had an increased risk of premenopausal breast cancer. No associations of the Bsm1 and Taq1 polymorphisms were observed in premenopausal women. Moreover, the Cdx2 GG (OR = 0.34, 95%CI = 0.192-0.602) genotype had a significant protective effect on breast cancer. However, strong LD was existed between Bsm1/Taq1 (D' = 0.757, CI = 0.67-0.82) and Apa1/Taq1 (D' = 0.695, CI = 0.6-0.77). Haplotype analysis showed no association between premenopausal breast cancer and VDR haplotypes. CONCLUSION: These analyses indicates that the VDR EcoRV A/A and Apa1 A/a genotypes may be risk factors for breast cancer development among premenopausal women.

Association of promoter region A-1012G polymorphism (rs4516035) of vitamin-D receptor gene with coronary artery disease.

OBJECTIVE: To investigate the correlation of Adenine-1012-Guanine (rs4516035) promoter region polymorphism of vitamin-D receptor gene with serum levels of omentin-1, vitamin-D and vitamin-D receptor protein in patients with coronary artery disease. METHODS: The case-control study was conducted from January to June 2020 at the cardiac unit of Civil hospital Karachi (CHK), and comprised coronary artery disease patients and controls. The tetra-primer amplification refractory mutation system polymerase chain reaction method was used to genotype Adenine-1210Guanine polymorphism in the vitamin D receptor gene. Serum levels of omentin-1, vitamin-D, and vitamin-D receptor protein were measured in both the groups using an enzyme-linked immunosorbent assay. Data was analysed using SPSS 17. RESULTS: Of the 1,000 subjects, there were 500(50%) cases; males 306(61.2%) and 194(38.8%) females with overall mean age of 51.08±9.55 years. The remaining 500(50%) were controls; 290(58%) males and 210(42%) females with overall mean age of 50.9±10.78 years. The mutant Guanine allele was more prevalent in controls 261(52.2%), and had a non-significant correlation with coronary artery disease (p=0.45). Among the cases, the wild Adenine-Adenine genotype had a higher prevalence 402(80.4%) and had a significant correlation with coronary artery disease (p<0.001). The heterozygous genotype Adenine-Guanine was significantly more predominant among the controls 346(69.2%) compared to the cases 66(13.2) (p=0.002). CONCLUSIONS: Adenine-1012-Guanine polymorphism in the vitamin-D receptor gene was found to be a protective polymorphism for coronary artery disease in the recessive model.

Suggested role of silent information regulator 1 (SIRT1) gene in female infertility: A cross-sectional study in Pakistan.

AIM & OBJECTIVE: Silent information regulator 1 (SIRT1) gene stimulates the expression of antioxidants and repairs damaged cells. It affects the mitochondrial activity within the oocytes to overcome the oxidant stress. We aimed to assess an association of SIRT1 polymorphism (Tag SNPs rs10509291 and rs12778366) with fertility, and assess serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), oestradiol, progesterone, manganese superoxide (MnSOD) and SIRT1. MATERIAL AND METHODS: In this cross-sectional study, 207 fertile and 135 infertile subjects between the ages of 18-45 years were recruited. Polymerase chain reaction (PCR) was performed; products were electrophoresed in a 2% agarose gel. Descriptive analysis of continuous variables was expressed as mean ± standard deviation. Mann-Whitney test was performed for comparison of groups, P value <.001 was considered significant. Single Nucleotide Polymorphism (SNP) data were analysed by applying chi-squared statistics. RESULTS: All subjects were age matched (P = .896). SIRT1 levels were significantly lower in infertile females when compared with fertile subjects (P < .001). AA (rs10509291) and CC (rs12778366) variant frequency was higher in the infertile than fertile subjects (P < .01). Similarly, the frequency of A allele (rs10509291) and C allele (rs12778366) was higher in infertile subjects (P < .001). Infertile females (29%) showed existence of SNP rs10509291 while 49% demonstrated genetic variation of rs12778366. MnSOD and SIRT1 levels were found to be lower in these subjects. CONCLUSION: The presence of SIRT1 genetic variants (rs10509291 and rs12778366) apparently disturbs the expression of SIRT1 deteriorating mitochondrial antioxidant function within the oocytes, instigating oxidative stress within. Their probable effect on modulating oocyte maturation may be the cause of infertility in females.

Association of BDNF and SERT gene polymorphisms with depression among Pakistani population.

Brain-derived neurotrophic factor (BDNF) and serotonin transporter (SERT) is implicated in the adverse life events which lead to depression. The variation in genetic make-up of BDNF (Val66Met) and SERT (5'-HTTLPR) are potential biomarkers in the development of neuropsychiatric disorders including depression. The purpose of this study was to investigate the correlation of functional polymorphisms of BDNF and SERT genes with depression among Pakistani population. A total of 373 participants (204 cases with depressive episodes and 169 healthy controls) with age between 14 and 65yrs were recruited from Pakistani population. BDNF and SERT gene polymorphisms were genotyped using PCR-RFLP analysis. The result showed that lack of association of Val66Met (χ2: 3.596, p>0.05) and 5'-HTTLPR (χ2: 0.634, p>0.05) gene polymorphisms were found with depression. However, SERT 'SL' (OR: 1.150, 95%CI: 0.601-2.201) and BDNF 'AA' (OR: 1.651, 95%CI: 0.585-4.660) and 'GA' (OR: 2.279, 95%CI: 0.825-6.298) genotypes might be a risk genotypes for depression. Hence, it is concluded that the functional BDNF (Val66Met) and SERT (5'-HTTLPR) gene polymorphisms may not be associated with depression. Replication studies on these polymorphisms with large sample size are needed.

Clearing the Clutter: Antiatherosclerotic activity of Eucalyptus camaldulensis crude extract.

Plant components have been extensively evaluated for their pharmacological activities. This study provides scientific rationale towards the therapeutic effect of Eucalyptus camaldulensis aqueous bark extract against induced atherosclerosis and hyperlipidemia in pigeons. Phytochemical components of Eucalyptus bark extract possess a great antioxidant activity that potentially reduced the risk of heart diseases. A total of 42 Pigeons of both sexes were distributed into negative control (fed normal grain diet), hyperlipidemic control (fed HFD 1% animal fat oil and 0.1% cholesterol for 3 months), test groups of variable doses (0.05, 0.1, 0.2 to 0.4 gms/kg BW for 21 days) and the group received atorvastatin daily after induction used. At the end of the experiment biochemical and histological evaluation has been performed. After HFD induction the serum levels of liver enzyme AST, glucose, urea, cholesterol, LDL, VLDL, and TG were significantly increased with the reduction in HDL levels. The atherogenic index was also found significantly raised. Microscopic examination of the liver and aorta showed the appearance of lipid-filled foam cells all over the liver parenchyma and intima after the HFD induction. Thus it was concluded that Eucalyptus aqueous bark extract can be effective against atherosclerosis and hyperlipidemia.

Association of oxidative stress with female infertility - A case control study.

OBJECTIVE: To compare stress markers and reproductive hormones in fertile and infertile females, and to relate the markers with age, duration and cause of infertility, and body mass index.. METHODS: The case-control study was conducted at Aga Khan University Hospital, Karachi, from March 2017 to February 2018. Females aged 16-50 years regardless of ethnic background were recruited from the Australian Concept Infertility Medical Centre, Karachi, and were equally divided into infertile cases group A, and fertile controls group B. Serum follicular stimulating hormone, luteinizing hormone, estradiol, glutathione reductase and cortisol were measured using enzyme-linked innmunosorbent assay. SPSS 19 was used for statistical analysis.. RESULTS: There were 328 female subjects divided into two equal groups of 164(50%). Serum luteinizing hormone and cortisol was higher in the group A than in group B (p<0.001). Serum glutathione reductase was low in group A compared to group B (p<0.001). Duration of infertility, serum levels of glutathione reductase and cortisol were also significantly different among infertile females when distributed on the basis of cause of infertility (p<0.05). Serum cortisol had negative correlation with glutathione reductase (p<0.001). Age and body mass index had a positive correlation with serum cortisol (p=0.035; p=0.63), while there was a negative correlation with glutathione reductase (p = -0.732).. CONCLUSIONS: Prolonged duration of infertility, age of females and body mass index enhanced the production of stress hormones and decreased antioxidant activity which augmented the risk of infertility.

Identification of Aspergillus flavus and aflatoxin in home mix layer poultry feed in relation to seasons in Karachi, Pakistan.

Fungal toxins in feed are leading issue in poultry industry causing a detrimental effect on the performance and health of poultry. The study was carried out to determine the incidence and concentration of the aflatoxins and their major producer Aspergillus flavus in home mix layer poultry feed in respect of seasonal variation throughout the year. A total of (n = 204) home mix poultry layer feed samples were analyzed for the isolation of fungi. The isolates were initially screened through colony morphology and microscopic examination. However, aflatoxin concentration was determined by ELISA. Revealed results indicated that, the highest percentage of A. flavus was found during the months of June to August 50/54 (92.5%) followed by September to November 43/65 (66.1%), March to May 21/40 (52.5%), and December to February 18/45 (40%). As a whole, the incidence was recorded 132/204 (64.7%). Moreover, of the 132 samples, 41 (31%) were exceeded in respect of aflatoxin contamination from the legal limit (20 µg/kg) imposed by Food Drug Association (FDA). Statistically, the growth of A. flavus and aflatoxin production was found significantly different in respect of seasonal variation. As highest total viable fungal count (9.9 × 104 CFU/g) and aflatoxin level (72.27 µg/kg) were recorded during the months of June to August and lowest in December to February. Consequently, instantaneous essential control measures are demanded regarding appropriate storage and adequate drying in post-harvesting season. Along with surveillance plans and austere regulations for monitoring the aflatoxin contents for the wellbeing of consumers.

Cytotoxic, embryotoxic, insecticidal and anti-microbial activities of standardized Areca catechu nut.

The study was aimed at evaluating various biological actions of widely consumed Areca catechu nut. The nut's ethanolic extract exhibited cytotoxicity (lung cancer cell line), embryotoxicity (chick embryo), phytotoxicity (Lemna minor), insecticidal (Rhyzopertha dominica), anti-bacterial (Pseudomonas aeruginosa), anti-fungal (Microsporum canis) and mitogenic (human blood lymphocytes) actions. The standardization results revealed presence of 1.7 µ g arecoline per mg of extract. In conclusion, the Areca nut is endowed with both harmful and beneficial biological actions. Keeping in view its wide consumption and ease of availability, the aforesaid information should be channelized for health and agricultural benefits.

Association between Vitamin D receptor (Cdx2, Fok1, Bsm1, Apa1, Bgl1, Taq1, and Poly (A)) gene polymorphism and breast cancer: A systematic review and meta-analysis.

The purpose of this systemic review and meta-analysis was to examine the relationship between VDR gene polymorphisms and breast cancer. Literature was searched through PubMed database, Google scholar, and the web of knowledge from December 2015 to January 2017 and consists of 34 studies (26,372 cases and 32,883 controls). All statistical measures were done using STATA version 11.2. The heterogeneity among studies was tested using I2 statistics. Mantel-Haenszel method and DerSimonian-Laird method were used to combine data from studies using both random-effect model and fixed-effect model, respectively. Potential publication bias was evaluated by Egger's test. Sensitivity analysis was also performed to evaluate the quality and consistency in results. The results of this meta-analysis revealed that VDR gene polymorphisms (Bsm1 bb vs BB; SOR = 1.18, 95% CI = 1.054-1.322, Apa1 aa vs AA; SOR = 1.18, 95% CI = 0.87-1.59, Poly (A) LL vs SS; SOR = 1.41, 95% CI = 1.06-1.88, Fok1 ff + Ff vs FF; SOR = 1.25, 95% CI = 0.896-1.759, Apa1 aa+Aa vs AA; SOR = 1.13, 95% CI = 0.95-1.35, Poly (A) LL + LS vs SS; SOR = 1.19, 95% CI = 1.00-1.43, Poly (A) L vs S; SOR = 1.18, 95% CI = 1.03-1.35) are associated with the breast cancer. Cdx2, Bgl1, and Taq1 do not show association with breast cancer. Thus, the finding of this meta-analysis concluded that VDR Bsm1, Apa1, Fok1, and Poly (A) gene polymorphisms may be susceptible for breast cancer development.

Gestational diabetes mellitus and the predisposing factors.

OBJECTIVE: To evaluate the occurrence of gestational diabetes mellitus and its association with demographic and anthropometric variables in pregnant women. METHODS: This cross-sectional study was conducted at the Aga Khan University Hospital, Abbasi Shaheed Hospital and Memon Hospital in Karachi, from February 2014 to December 2015, and comprised pregnant women who were screened by 75-g 2-hour oral glucose tolerance test, (24-28 weeks of gestation) and classified as per the criteria of the International Association of Diabetes and Pregnancy Study Group. Weight, body mass index and serum glycated haemoglobin levels were measured. Women with pre-gestational diabetes were excluded. SPSS 21 was used for data analysis. RESULTS: Of the 1,210 participants, 208(17.2%) had gestational diabetes, while 1,002(82.8%) did not have the condition. Gestational diabetes was associated with advancing age, deranged glycated haemoglobin, elevated body mass index at booking (p<0.001) and history of first-degree type 2 diabetic relatives (p=0.05). When stratified according to ethnicities, no difference was observed in terms of gestational diabetes predilection among those who had the condition (p>0.05). CONCLUSIONS: Pre-existing adiposity and presence of strong family history rendered a considerable number of pregnant women to suffer from gestational diabetes.

KCNQ1 rs2237895 polymorphism is associated with Gestational Diabetes in Pakistani Women.

BACKGROUND AND OBJECTIVE: Genetic studies on gestational diabetes (GDM) are relatively scarce; moreover, limited data is available for KCNQ1 polymorphism in Pakistani pregnant women. We aimed to determine the frequency of KCNQ1 rs2237895 in GDM and normal pregnant controls and its association with GDM-related phenotypes. METHODS: A total of 637 pregnant females (429 controls and 208 cases) in their second trimester were classified according to the International Association of the Diabetes and Pregnancy Study criteria in this study. Their blood samples were genotyped for KCNQ1 SNP rs2237895 using PCR-RFLP method and sequencing. Fasting and two hour-post glucose load blood levels, serum HbA1c, insulin, and anthropometric assessment was performed.: Pearson's Chi Square test, Mann- Whitney U test, and regression analyses were performed. A p-value of <0.05 was considered significant. RESULTS: The variant genotyped was in Hardy-Weinberg equilibrium (p>0.05). The rs2237895 showed an association with GDM (OR 2.281; 1.388-3.746: p <0.001) and remained significant after multiple adjustments for age and body mass index (OR 2.068; 1.430-2.997: p=0.005). The C allele showed positive association with insulin level, and HOMA-IR in study subjects. CONCLUSIONS: This study identifies that KCNQ1 rs2237895 polymorphisms might be associated with risk of GDM in Pakistani population and that it is related to higher glucose levels and insulin resistance. Further large scale studies are required to consolidate on the functional aspect of this polymorphism.

Correlation of ABH blood group antigens secretion with Helicobacter pylori infection in Pakistani patients.

OBJECTIVES: A and B blood group antigens are fucosylated carbohydrate present on human erythrocytes and body secretions. Their presence in body secretions depends on the expression of a dominant allele of secretor gene FUT2 and is correlated with susceptibility to various infectious and non-infectious diseases. We investigated the correlation of blood group and ABH antigen secretion with Helicobacter pylori infection and gastroduodenal symptoms and analysed the distribution of babA gene among ABH secretors and non-secretors. METHODS: Two hundred and ninety patients who underwent gastroduodenal endoscopy during 2011 to 2012 participated. Gastric biopsy, saliva and blood samples were obtained from every patient. Gastric biopsies were subjected to rapid urease test and PCR for the detection of H. pylori and babA gene. Blood grouping and ABH antigens secretions were determined by Lewis blood group phenotyping and haemagglutination inhibition test. RESULTS: 50.34% of patients were ABH antigen secretors and 45.51% non-secretors. Distribution analysis of blood group revealed that 40 blood group B, 67 blood group A 20 blood group O and 19 blood group AB patients secreted ABH antigens in saliva. Fifty-six blood group O, 19 blood group B, 32 blood group A and 17 blood group AB patients were non-secretors. Gastroduodenal complaints were common among non-secretors. Sixty-two percent of patients with a combination of duodenal ulcer and gastro-oesophageal reflux and 54% of patients with gastritis were non-secretors. Of 290 samples, 31.02% were positive for H. pylori. Thirty percent of these tested positive for babA gene; the majority belonged to non-secretor blood group O. CONCLUSIONS: Our results suggest that the infection of H. pylori is correlated with ABO blood groups and blood group antigens secretion in body fluids.

Effect of endometrial thickness on pregnancy outcome after intracytoplasmic sperm injection.

OBJECTIVE: To identify cut-off value of endometrial thickness required for implantation of embryo after intracytoplasmic sperm injection. METHODS: The auasi-experimental study was conducted from July 2011 to June 2012 at an assisted reproductive clinic in Islamabad. Down-regulation of ovaries, controlled ovarian stimulation, oocyte pick-up, in vitro fertilisation, blastocyst transfer and confirmation of pregnancy with beta human chorionic gonadotropin more than 5mIU/ml. Patients were categorised into two groups on the basis of endometrial thickness < 8mm and >8mm.On ovulation induction, before human chorionic gonadotropin injection, endometrial thickness was measured by trans-vaginal scan. Receiver operating curve was used to define groups on the basis of endometrial thickness cut-off value for pregnancy. The groups were compared in terms of the number of retrieved, mature and fertilised oocytes along with oocyte maturity, fertilisation and implantation rates by chi square test. RESULTS: There were 282 females; 116(41%) in Group A with endometrial thickness < 8mm, and 166(59%) in Group B with endometrial thickness >8mm. In group A, 6(5%) and in Group B, 95(57.2%) patients had a positive pregnancy test. The number of mature, fertilised oocytes and cleaved embryos was significantly high in Group B (p=0.01; p=0.001; p=0.001respectively). Increase in endometrial thickness enhanced chances of oocyte maturity, fertilisation, cleavage and implantation (p< 0.0001 each). CONCLUSIONS: Endometrial thickness of 8mm was associated with a positive pregnancy outcome after intracytoplasmic sperm injection. Implantation of embryo was facilitated by better oocyte parameters, oocyte maturity, fertilisation and its cleavage in females who exhibited endometrial thickness above the cut-off value.

Interplay of "leukemia inhibitory factor receptor gene" (rs3099124) polymorphism, leukemia inhibitory factor and ovarian steroids with unexplained infertility.

OBJECTIVE OF THE STUDY: To explore the association of leukemia inhibitory factor receptor (LIFR) gene variant rs3099124, ovarian steroids, and leukemia inhibitory factor with unexplained infertility in Pakistani females. METHODOLOGY: A case-control investigation in which eighty-one (81) females with unexplained infertility and one hundred and sixty-two (162) fertile counterparts (age and body mass index compared) were recruited between October 2016 and 2018. Ten milliliters of venous blood was collected from all participants. "Genomic DNA" was taken out from lymphocytes in peripheral blood samples. "Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS-PCR)" was constructed through software "Primer-I". Amplification was carried out by "T-ARMS-PCR" followed by subsequent sequencing for confirmation and extensive consonance. Estradiol, Progesterone and Leukemia Inhibitory Factor (LIF) were measured in serum by ELISA. RESULTS: Statistically significant difference was noticed in genotype frequency in "LIFR-gene variant; rs3099124" (χ2 = 28.222, P value < 0.01) between research participants. Although, rs "3099124" "AA" (OR = 0.000; 95%CI = 0-0) and "GA" genotypes (OR = 0.525; 95%CI = 0.226-1.22) showed non-significant safety/protection against unexplained infertility yet minor/risk allele "A" frequency was greater in women with unexplained infertility suggesting a possible explanation of implantation failure. LIF concentration varied between fertile and infertile groups (χ2 = 9.857, P < 0.05) revealing significant threat of unexplained infertility in women with decreased LIF concentration (OR = 2.316, 95%CI = 1.214-4.416). Progesterone was significantly related to unexplained infertility in both study groups (χ2 = 20.347, P < 0.05). High progesterone reduced the possibility of unexplained infertility (OR = 0.306; 95% CI = 0.166-0.567). CONCLUSION: LIFR gene variation (rs3099124) and reduced LIF secretion may cause implantation failure in women with unexplained infertility.

Colloidal gold based immunochromatographic detection of Mycoplasmopsis synoviae infection and its prevalence in avian species of Karachi, Pakistan.

Avian mycoplasmosis is an infection that commonly prevails in birds, particularly in poultry chickens. Among mycoplasmosis causing organisms, Mycoplasmopsis synoviae is a predominant and lethal pathogen to the aves. Considering the increased incidence of infections by M. synoviae, the prevalence of M. synoviae was deduced in poultry chickens and fancy birds of Karachi region. The lungs and tracheal samples from chicken and dead fancy birds and swab samples from live fancy birds were collected and investigated by amplifying 16 s rRNA gene of M. synoviae. Biochemical characteristics of M. synoviae was also evaluated. Furthermore, surface-associated membrane proteins, that represent key antigens for diagnosis of M. synoviae infection was extracted by Triton X- 114 method. Results showed that M. synoviae was detected more frequently in lungs than in trachea, that could be due to its invasion capacity and tissue affinity. SDS PAGE analysis of extracted membrane proteins showed two prominent hydrophobic proteins of different molecular mass including proteins of 150 and 50 kDa. Protein of 150 kDa was purified by size exclusion chromatography and it exhibited agglutinogen activity. Purified protein was used in the development of one-step immunochromatographic (ICT) assay for the detection of antibodies against M. synoviae using gold nanoparticles coated with polyclonal antibodies. Low levels of antibodies were detected by the developed ICT kit, which has 88% sensitivity with 92% specificity.

Association of Visfatin gene polymorphism with obesity related metabolic disorders among Pakistani population: a case control study.

In recent years, the global prevalence of obesity and its associated metabolic disorders has reached alarming levels, presenting a significant challenge to public health worldwide. Visfatin, also known as pre-B cell colony-enhancing factor (PBEF) or nicotinamide phosphoribosyltransferase (NAMPT), is an adipokine that has been implicated in various physiological processes, including glucose homeostasis, lipid metabolism, and inflammation. The main objective of this proposed study is to find out the association between visfatin genetic variants and metabolic syndrome. The sample size of the study consisted of 300 blood samples (150 control and 150 cases). This study found that the genotypic frequency of visfatin SNPs, including rs2302559 (OD: 18.222; 95% CI 10.228-32.466; p-value < 0.001) and rs1215113036 (OD: 129.40; 95% CI 44.576-375.693; p-value < 0.001) were significantly associated with metabolic syndrome. Moreover, the frequency of the mutant alleles of both visfatin SNPs was found to be higher in patients with metabolic syndrome as compared to controls. Results of the current study indicate that people with any genetic variation of Visfatin, such as rs2302559 and rs1215113036, are more likely to develop metabolic syndrome. Visfatin genetic variants are linked to an increased risk of metabolic syndrome, implying it's role in disease pathophysiology.

Characterization of H5N1 highly pathogenic avian influenza viruses isolated from poultry in Pakistan 2006-2008.

Nine avian influenza viruses (AIV), H5N1 subtype, were isolated from dead poultry in the Karachi region of Pakistan from 2006 to 2008. The intravenous pathogenicity indices and HA protein cleavage sites of all nine viruses were consistent with highly pathogenic AIV. Based on phylogenetic analysis of the HA genes, these isolates belong to clade 2.2 and both the HA and NA are closely related to each other (nucleotide identities above 99.0%) and to other Middle Eastern H5N1 AIV isolates (nucleotide identities above 98.0%). The phylogenetic data suggest that the virus in both epornitics of H5N1 HPAIV in commercial poultry in the Karachi region of Pakistan between 2006 and 2008 were from a very closely related source, however, there is inadequate epidemiological data to determine what the reservoir was for the virus between the 2006 and 2007 outbreaks other than that there was a single introduction into the region.

Sesquiterpene from Polygonum barbatum disrupts mitochondrial membrane potential to induce apoptosis and inhibits metastasis by downregulating matrix metalloproteinase and osteopontin in NCI-H460 cells.

Globally, lung cancer accounts for 18% of cancer-associated mortalities. Among the subtypes, non-small cell lung cancer (NSCLC) is the most prevalent. The increased resistance and poor survival rates signify disease aggressiveness and thus require a search for an alternative anticancer molecule. Earlier, the sesquiterpene, i.e., compound 3 ((E)-methyl 6-acetoxy-7-methoxy-1-(2-methylpropylidene)-1H-indene-3-carboxylate) from Polygonum barbatum, was isolated, characterized by us, and reported for preliminary anticancer activity. Therefore, based on these results, this study was designed to explore the underlying molecular mechanism of apoptosis and metastasis against NCI-H460 cells. The molecular mechanism of compound 3 inducing apoptosis and inhibiting metastasis was elucidated by analyzing mitochondrial membrane potential, DNA fragmentation, clonogenic assay, invasion assay, and expression of apoptotic (caspases 3, 6, 8, 9, and BAK) and metastatic markers (MMP 2, MMP 9, and osteopontin). Compound 3 significantly inhibited cell proliferation and induced apoptosis via the intrinsic route, i.e., the mitochondrial pathway, by disrupting mitochondrial membrane potential. The enhanced expression of caspases 6, 9, BAK, and HRK with downregulation of Bcl-2L1 and Ki67 further confirmed the involvement of the intrinsic apoptotic pathway. Moreover, compound 3 restricted the invasive nature of NCI-H460 cells evinced by reduced cell invasion in Boyden chamber invasion assay and downregulating the expression of metastatic markers, i.e., matrix metalloproteinase 2/9 and VEGF. It was also found to block osteopontin by negatively regulating its expression, a marker protein in cancer management. Conclusively, this sesquiterpene exhibited potent anticancer and antimetastatic activity and can be explored further as possible pharmacophores.

Interplay between oxidative stress, SIRT1, reproductive and metabolic functions.

Silent information Regulators (SIRT1) gene stimulates antioxidants' expression, repairs cells damaged by oxidative stress (OS), and prevents the cells' dysfunction. In particular, the role of different Sirtuins, particularly SIRT1 in reproduction, has been widely studied over the past decade. Decreased SIRT 1 causes mitochondrial dysfunction by increasing Reactive Oxygen Species (ROS), lipid peroxidation, and DNA damage in both male and female gametes (Sperms and Oocytes), leading to infertility. In the female reproductive system, SIRT1 regulates proliferation and apoptosis in granulosa cells (GCs), and its down-regulation is associated with a reduced ovarian reserve. SIRT1 also modulates the stress response to OS in GCs by targeting a transcription factor vital for ovarian functions and maintenance. ROS-mediated damage to spermatozoa's motility and morphology is responsible for 30-80% of men's infertility cases. High levels of ROS can cause damage to deoxyribo nucleic acid (DNA) in the nucleus and mitochondria, lipid peroxidation, apoptosis, inactivation of enzymes, and oxidation of proteins in spermatozoa. SIRT 1 is a cardioprotective molecule that prevents atherosclerosis by modulating various mechanisms such as endothelial injury due to impaired nitric oxide (NO) production, inflammation, OS, and regulation of autophagy. SIRT 1 is abundantly expressed in tubular cells and podocytes. It is also found to be highly expressed in aquaporin 2 positive cells in the distal nephron suggesting its involvement in sodium and water handling. SIRT1 improves insulin resistance by reducing OS and regulating mitochondrial biogenesis and function. It also decreases adiposity and lipogenesis and increases fatty acid oxidation. So, its involvement in the multiple pathways ensures its unique role in reproductive and metabolic derangement mechanisms.