ANTICARCINOGENIC ACTIVITY OF RICE BRAN PHYTIC ACID AGAINST HUMAN BREAST CANCER CELL LINE (MCF-7)

Phytic acid (PA) has been reported for anti-inflammatory, antioxidant and anticancer activity. However, molecular mechanism of anticancer activity is not clear. This study investigated the anticancer activity of rice bran PA against breast cancer (MCF-7). Cytotoxicty of PA (0 to 7 mM) against MCF-7 cells was examined by MTT and LDH assays after 24 and 48 h treatment. Apoptotic activity was evaluated by expressional analysis of apoptosis-regulatory genes [i.e., p53, Bcl-2, Bax, caspase-3 and -9] by reverse transcriptase-PCR and DNA fragmentation assay. PA inhibited the growth of MCF-7 cells in a concentration dependent manner (p≤0.04). After 48 h treatment, cells viability was recorded 80.9, 71.1, 59.8, 36.6, 26.7 and 15.9% in MTT assay and 85.3, 72.6%, 62.3%, 42.1, 31.7 and 21.7% in LDH assay at concentration of 1.4, 2.2, 3.0, 3.8, 4.6, and 5.4 mM respectively. Hence, treatment of PA for 24 h, recorded viability of cells 84.6, 73.8, 61.0, 47.0, 28.8 and 17.3% in MTT assay and 87.8, 77.5%, 62.9%, 49.8, 35.7 and 23.3% in LDH assay at concentration of 2, 3, 4, 5, 6, and 7 mM, respectively. PA treated MCF-7 cells showed up-regulation of p53, Bax, caspase-3 and -9, and down-regulation of Bcl-2 gene (p ≤0.03). At IC50 (3.4 mM) of PA, the p53, Bax, caspase 3 and -9 genes were up-regulated by 6.34, 4.90, 23.45 and 15.03 folds respectively. Also, the fragmented genomic DNA in PA treated cells showed the signs of apoptosis. Our study endorsed the biological activity of PA and demonstrated the PA induced growth inhibition and apoptosis in MCF-7 cells by modulating the expression of apoptosis-regulatory genes. Keyword: Phytic acid, antioxidant, cytotoxicity, apoptosis, caspases, p53, Bax, Bcl-2, DNA fragmentatio

Prevalence of Self-Medication of Psychoactive Stimulants and Antidepressants among Undergraduate Pharmacy Students in Twelve Pakistani Cities

Purpose: To evaluate the prevalence of self-medication of psychoactive stimulants and antidepressants among pharmacy students of Pakistan.Methods: A cross-sectional survey on self-medication of psychoactive stimulants and antidepressants among pharmacy students was conducted with a structured and validated questionnaire distributed to a total of 2981 final year undergraduate pharmacy students in 12 major Pakistani cities (Karachi, Lahore, Islamabad, Rawalpindi, Sargodha, Dera Ismail Khan, Abbottabad, Bahawalpur, Hyderabad, Faisalabad, Multan and Peshawar) of Pakistan. Out of this, 2516 (718 male and 1798 female) students completed and returned the questionnaire.Results: Prevalence of self-medication of psychoactive stimulants was 1.31 (1.13 – 1.75 for 95% CI) and antidepressants was 8.34 (8.03 – 8.85 for 95% CI). A majority of the students (63 %) identified academic competition as a driving force for indulging in self-medication of psychoactive stimulants while nearly all the students (96 %)admitted using antidepressants to obtain relief from the pressure of studies (p < 0.05).Conclusion: Pakistani pharmacy students, despite being aware of the hazards of psychoactive stimulants, indulge in self-medication. Prevalence of self-medication with antidepressants is very high among the students due to the pressure of studies. Primarily, academic competition is the major driving force for the use of psychoactive stimulants.Keywords: Self-medication, Psychoactive stimulants, Antidepressants, Pharmacy students, Academicpressur

Development and Validation of a Deep Learning-Based Microsatellite Instability Predictor from Prostate Cancer Whole-Slide Images

Microsatellite instability-high (MSI-H) is a tumor agnostic biomarker for immune checkpoint inhibitor therapy. However, MSI status is not routinely tested in prostate cancer, in part due to low prevalence and assay cost. As such, prediction of MSI status from hematoxylin and eosin (H&E) stained whole-slide images (WSIs) could identify prostate cancer patients most likely to benefit from confirmatory testing and becoming eligible for immunotherapy. Prostate biopsies and surgical resections from de-identified records of consecutive prostate cancer patients referred to our institution were analyzed. Their MSI status was determined by next generation sequencing. Patients before a cutoff date were split into an algorithm development set (n=4015, MSI-H 1.8%) and a paired validation set (n=173, MSI-H 19.7%) that consisted of two serial sections from each sample, one stained and scanned internally and the other at an external site. Patients after the cutoff date formed the temporal validation set (n=1350, MSI-H 2.3%). Attention-based multiple instance learning models were trained to predict MSI-H from H&E WSIs. The MSI-H predictor achieved area under the receiver operating characteristic curve values of 0.78 (95% CI [0.69-0.86]), 0.72 (95% CI [0.63-0.81]), and 0.72 (95% CI [0.62-0.82]) on the internally prepared, externally prepared, and temporal validation sets, respectively. While MSI-H status is significantly correlated with Gleason score, the model remained predictive within each Gleason score subgroup. In summary, we developed and validated an AI-based MSI-H diagnostic model on a large real-world cohort of routine H&E slides, which effectively generalized to externally stained and scanned samples and a temporally independent validation cohort. This algorithm has the potential to direct prostate cancer patients toward immunotherapy and to identify MSI-H cases secondary to Lynch syndrome

'You were quiet - I did all the marching': Research processes involved in hearing the voices of South Asian girls

This article is available open access through the publisher’s website at the link below. Copyright @ 2011 A B Academic Publishers.This article provides insights into the outcomes of reflection following two interview approaches used to explore narratives of the lived, individual experiences of South-Asian girls living in West London. In attempting to illuminate and re-present the cultural experiences as told by these girls, the choice of interview approach became critical in allowing the voices to be effectively heard (Rogers, 2005). This article therefore considers how a semi-structured interview approach offered valuable insights into the girls' experiences but became constraining for both researcher and participant in unveiling the complexity and depth of their lives. These constraints emerged through reflection by both participants and researcher. As a result of reflexivity during the research process, the researcher moved towards the use of research conversations during the second phase of the study. Ultimately the study revealed how the girls felt empowered by the opportunity to narrate their individual experiences and tell of their lives. In narrating their reflections on being part of the research, there was a clear recognition that the process facilitated the articulation of new voices and ‘multi-voicedness’ (Moen, 2006

Genetic variability in the precore and core promoter regions of hepatitis B virus strains in Karachi

BACKGROUND: Hepatitis B virus (HBV) genotypes have distinct geographic distribution. Moreover, much genetic variability has been described in the precore (PC) and basal core promoter (BCP) regions of the HBV genome. The local prevalence of HBV genotypes and mutations has not been well studied. The aim of the present study is to determine the prevalence of HBV genotypes and mutations in the PC and BCP region in HBV strains in Karachi. METHODS: A total of 109 chronic hepatitis B patients with detectable HBV DNA by a PCR assay were enrolled in the study. Sera were tested for HBeAg, anti-HBe antibody and liver profile. HBV genotypes and mutations in the PC and BCP regions were detected by INNO-LiPA line-probe assays. RESULTS: Of the 109 patients investigated, 38 (35%) were HBeAg positive while 71 (65%) were HBeAg negative. Genotype D was present in 100% of the patients. Two patients had co-infection with genotype A. There was no significant difference in the baseline characteristics, mean ALT levels, and presence of clinical cirrhosis in patients with HBeAg positive or negative strains with or without PC and BCP mutations. Of the 38 HBeAg positive patients, 9 (24%) had PC and BCP mutations. In the HBeAg negative patient group, mutations were detected in 44 (62%) of the strains investigated. More than one mutation was common, seen in 26 (37%) patients with HBeAg negative disease and 6 (16%) patients with HBeAg positive disease. Twelve (17%) HBeAg negative patients had dual T1762 and A1764 mutations. None of the HBeAg positive patients had T1762 mutation. Mutations were undetectable in 27 (38%) of patients with HBeAg negative disease. CONCLUSION: Our study shows that type D is the main HBV genotype in Karachi, Pakistan. Significant numbers of patients infected with this genotype have PC and BCP variants. Mutations at more than one site are common. Patients harboring these mutants do not differ significantly in their clinical presentation from patients having wild type infection

An unbiased lipid phenotyping approach to study the genetic determinants of lipids and their association with coronary heart disease risk factors

Direct infusion high-resolution mass spectrometry (DIHRMS) is a novel, high-throughput approach to rapidly and accurately profile hundreds of lipids in human serum without prior chromatography, facilitating in-depth lipid phenotyping for large epidemiological studies to reveal the detailed associations of individual lipids with coronary heart disease (CHD) risk factors. Intact lipid profiling by DIHRMS was performed on 5662 serum samples from healthy participants in the Pakistan Risk of Myocardial Infarction Study (PROMIS). We developed a novel semi-targeted peak-picking algorithm to detect mass-to-charge ratios in positive and negative ionization modes. We analyzed lipid partial correlations, assessed the association of lipid principal components with established CHD risk factors and genetic variants, and examined differences between lipids for a common genetic polymorphism. The DIHRMS method provided information on 360 lipids (including fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, and sterol lipids), with a median coefficient of variation of 11.6% (range: 5.4–51.9). The lipids were highly correlated and exhibited a range of associations with clinical chemistry biomarkers and lifestyle factors. This platform can provide many novel insights into the effects of physiology and lifestyle on lipid metabolism, genetic determinants of lipids, and the relationship between individual lipids and CHD risk factors

Human knockouts and phenotypic analysis in a cohort with a high rate of consanguinity

A major goal of biomedicine is to understand the function of every gene in the human genome. Loss-of-function mutations can disrupt both copies of a given gene in humans and phenotypic analysis of such 'human knockouts' can provide insight into gene function. Consanguineous unions are more likely to result in offspring carrying homozygous loss-of-function mutations. In Pakistan, consanguinity rates are notably high. Here we sequence the protein-coding regions of 10,503 adult participants in the Pakistan Risk of Myocardial Infarction Study (PROMIS), designed to understand the determinants of cardiometabolic diseases in individuals from South Asia. We identified individuals carrying homozygous predicted loss-of-function (pLoF) mutations, and performed phenotypic analysis involving more than 200 biochemical and disease traits. We enumerated 49,138 rare (<1% minor allele frequency) pLoF mutations. These pLoF mutations are estimated to knock out 1,317 genes, each in at least one participant. Homozygosity for pLoF mutations at PLA2G7 was associated with absent enzymatic activity of soluble lipoprotein-associated phospholipase A2; at CYP2F1, with higher plasma interleukin-8 concentrations; at TREH, with lower concentrations of apoB-containing lipoprotein subfractions; at either A3GALT2 or NRG4, with markedly reduced plasma insulin C-peptide concentrations; and at SLC9A3R1, with mediators of calcium and phosphate signalling. Heterozygous deficiency of APOC3 has been shown to protect against coronary heart disease; we identified APOC3 homozygous pLoF carriers in our cohort. We recruited these human knockouts and challenged them with an oral fat load. Compared with family members lacking the mutation, individuals with APOC3 knocked out displayed marked blunting of the usual post-prandial rise in plasma triglycerides. Overall, these observations provide a roadmap for a 'human knockout project', a systematic effort to understand the phenotypic consequences of complete disruption of genes in humans.D.S. is supported by grants from the National Institutes of Health, the Fogarty International, the Wellcome Trust, the British Heart Foundation, and Pfizer. P.N. is supported by the John S. LaDue Memorial Fellowship in Cardiology from Harvard Medical School. H.-H.W. is supported by a grant from the Samsung Medical Center, Korea (SMO116163). S.K. is supported by the Ofer and Shelly Nemirovsky MGH Research Scholar Award and by grants from the National Institutes of Health (R01HL107816), the Donovan Family Foundation, and Fondation Leducq. Exome sequencing was supported by a grant from the NHGRI (5U54HG003067-11) to S.G. and E.S.L. D.G.M. is supported by a grant from the National Institutes of Health (R01GM104371). J.D. holds a British Heart Foundation Chair, European Research Council Senior Investigator Award, and NIHR Senior Investigator Award. The Cardiovascular Epidemiology Unit at the University of Cambridge, which supported the field work and genotyping of PROMIS, is funded by the UK Medical Research Council, British Heart Foundation, and NIHR Cambridge Biomedical Research Centre ... Fieldwork in the PROMIS study has been supported through funds available to investigators at the Center for Non-Communicable Diseases, Pakistan and the University of Cambridge, UK