Identification of specific codon 201 mutation of the DCC Gene in the colonoscopic specimen of colorectal cancer.

Objective: To identify the mutation in codon 201 of the deleted in colorectal cancer gene in colorectal cancer, and to correlate that mutation to the histopathological grading of colorectal cancer. METHODS: The cross-sectional study was conducted from February 2019 to February 2021 after approval from the ethics review board of the Dow University of Health Sciences, Karachi, and comprised biopsy-proven colorectal cancer patients regardless of age and gender. After histopathological reporting, formalin-fixed paraffin-embedded tissue blocks of colorectal cancer were used for deoxyribonucleic acid extraction, followed by polymerase chain reaction optimisation and deoxyribonucleic acid Sanger sequencing for mutational analysis. Data was analysed using SPSS 25. RESULTS: Of the 100 biopsy specimens assessed, 45(45%) were selected. Of them, 13(29%) samples failed to show any band on gel electrophoresis. The remaining 32(71%) samples were used for Sanger sequencing. Of these, 1(3%) sample did not sequence, while 31(97%) showed sequencing. All the sequenced samples identified a mutation in codon 201 of exon 3 in the deleted in colorectal cancer gene; 30(97%) showed homozygosity, and 1(3%) showed heterozygosity. No significant association of point mutation was noted with various demographic and clinicopathological parameters (p>0.05). Conclusion: The deleted in colorectal cancer gene's missense mutation in codon 201 was frequently observed in colorectal cancer patients.

The relationship of maternal age, BMI, serum Tenascin-C, and HOMA-IR values with gestational diabetes mellitus.

Objectives: To explore the relationship, if any, of gestational diabetes mellitus with maternal age, body mass index, serum tenascin-C and homeostatic model assessment for insulin resistance, and to see if these could act as predictive markers for gestational diabetes mellitus. METHODS: The case-control study was conducted from February to August 2022 at the outpatient department of gynaecology and obstetrics at the Civil Hospital, Karachi, and comprised pregnant females aged 18-40 years having gestational age 20-34 weeks. After noting down baseline characteristics and anthropometric measurements, the participants were subjected to oral glucose tolerance test on the basis of which they were divided into three groups; pregnant healthy controls in group 1, those with gestational diabetes mellitus on diet control in group 2, and those with gestational diabetes mellitus taking medicines for the condition in group 3. Fasting serum samples were used for further analysis using enzyme-linked immunosorbent assay kits. Data was analysed using SPSS 21. RESULTS: Of the 90 subjects, 30(33.3%) were in group 1 with mean age 26.0±4.9 years, 30(33.3%) were in group 2 with mean age 30.7±5.6 years, and 30(33.3%) were in group 3 with mean age 29.1±5.5 years. Age, gestational age, body mass index and homeostatic model assessment for insulin resistance values were significantly higher in groups 2 and 3 compared to group 1 (p<0.05), while serum Tenascin-C values were not significantly different (p>0.05). CONCLUSIONS: HOMA-IR values and BMI were more reliable in diagnosing GDM before its onset, and should be included in the screening test for GDM in early pregnancy.

Transcript-Level In Silico Analysis of Alzheimer's Disease-Related Gene Biomarkers and Their Evaluation with Bioactive Flavonoids to Explore Therapeutic Interactions.

Alzheimer's disease (AD) is a progressive brain disorder that can significantly affect the quality of life. We used a variety of in silico tools to investigate the transcript-level mutational impact of exonic missense rare variations (single nucleotide polymorphisms, SNPs) on protein function and to identify potential druggable protein cavities that correspond to potential therapeutic targets for the management of AD. According to the NIA-AA (National Institute on Aging-Alzheimer's Association) framework, we selected three AD biomarker genes (APP, NEFL, and MAPT). We systematically screened transcript-level exonic rare SNPs from these genes with a minor allele frequency of 1% in 1KGD (1000 Genomes Project Database) and gnomAD (Genome Aggregation Database). With downstream functional effect predictions, a single variation (rs182024939: K > N) of the MAPT gene with nine transcript SNPs was identified as the most pathogenic variation from the large dataset of mutations. The machine learning consensus classifier predictor categorized these transcript-level SNPs as the most deleterious variations, resulting in a large decrease in protein structural stability (ΔΔG kcal/mol). The bioactive flavonoid library was screened for drug-likeness and toxicity risk. Virtual screening of eligible flavonoids was performed using the MAPT protein. Identification of druggable protein-binding cavities showed VAL305, GLU655, and LYS657 as consensus-interacting residues present in the MAPT-docked top-ranked flavonoid compounds. The MM/PB(GB)SA analysis indicated hesperetin (-5.64 kcal/mol), eriodictyol (-5.63 kcal/mol), and sakuranetin (-5.60 kcal/mol) as the best docked flavonoids with the near-native binding pose. The findings of this study provide important insights into the potential of hesperetin as a promising flavonoid that can be utilized for further rational drug design and lead optimization to open new gateways in the field of AD therapeutics.

Comparison Of Potential Plasma Biomarkers In Alzheimer's Disease And Neurodegenerative Dementias In Pakistani Population.

This study aimed to compare the mean plasma levels of Amyloid ß42, Phosphorylated Tau and Neurofilament Light chain in patients diagnosed with Alzheimer's Clinical Syndrome (ACS), and other neurodegenerative dementias to find affordable and less-invasive means of diagnosing Alzheimer's disease (AD) early in its course. Blood samples of 36 subjects presenting with cognitive decline to the neurology OPDs of Dow and Civil hospitals, Karachi, were centrifuged, and plasma was stored at -80℃. Before analysis, it was thawed at 4℃ and protein levels were measured through ELISA. Two-thirds of the patients were females but age distribution across both the groups was not significantly different (p=0.21). No difference was observed in the mean plasma concentrations of Aß42, P-Tau, and NFL between the two groups (p-values 0.78 and 0.27 and 0.09 respectively). Our study suggests that despite being promising in CSF, Aß42, P-Tau, and NFL cannot differentiate between different neurodegenerative dementias when measured in plasma.

Confronting Challenges: An Inductive Thematic Analysis Of Barriers And Solutions To Undergraduate Medical Research In Pakistan.

OBJECTIVE: To qualitatively analyse the barriers in pursuing undergraduate research, and to propose solutions for the problems identified. METHODS: The qualitative study was conducted from May to October 2021 after approval from the institutional ethics review board of the Dow University of Health Sciences, Karachi, and comprised undergraduate students of either gender at various public and private medical universities across Pakistan having some level of research experience and good communication skills. Data triangulation was employed to collect qualitative data through open-ended survey, face-to-face interviews and focus group discussions. Using the information of one method to inform the rest, linked trajectories were established that allowed validation of information at each level. Data was coded manually by two researchers independently. Data was subjected to inductive thematic analysis. RESULTS: Of the 33 subjects, 17(51.5%) were males, 17(51.5%) were from private medical colleges, 18(54.5%) were from Karachi, and 11(33.3%) were in the final year of medical school. Overall, 13(39.4%) students completed the open-ended survey, 6(18.2%) completed face-to-face interviews, and 14(42.4%) participated in focus group discussions. Thematic analysis showed that students were interested in research to improve their career prospects, but not all were passionate about it. Students were not satisfied with the quality of research being conducted in the country. Dearth of motivated faculty, unavailability of well-maintained and digitalised data registries, ineffective research methodology teaching and lack of access to medical journals and research software were the major barriers in undergraduate research. Time constraint was a projecting problem which challenged the students. Frequent research workshops and conferences, strong networking, reorienting curriculum to provide early exposure to research and student-led initiatives were suggested to improve undergraduate research in Pakistan. CONCLUSIONS: Students' lack of initiative coupled with administrative and faculty-related issues pose a serious threat to the future of evidence-based medicine. Proposed solutions offer a ray of hope to the future of undergraduate research in Pakistan.

Plasma levels of phosphorylated tau and neurofilament light chain as potential biomarkers for Alzheimer's disease: A biochemical analysis in Pakistani population.

The National Institute on Aging-Alzheimer's Association's research framework in 2018 proposed a molecular construct for the diagnosis of Alzheimer's disease (AD). Nonetheless, the clinical exclusionary strategy is still the mainstay of AD diagnosis in Pakistan. We looked at the plasma levels of amyloid beta-42 (Aß-42), phosphorylated tau (P-tau), and neurofilament light (NFL) in patients with Alzheimer's clinical syndrome (ACS) and healthy controls (HC) from the Pakistani population to keep pace with the global efforts towards establishing accessible and affordable biochemical diagnostic markers for AD in Pakistan. Consultant neurologists screened patients who presented with cognitive impairment to three large tertiary care hospitals in Karachi, and after receiving informed consent, recruited participants with ACS and HC from the same facilities. We collected 5cc of blood in EDTA tubes along with demographic and lifestyle information of the subjects. Plasma aliquots were stored at -80°C after centrifugation. For analysis it was thawed at 4℃ and levels of the three proteins were measured through ELISA. Data from 28 ACS patients and 28 age matched healthy controls were evaluated. Among demographic factors, education and depression were related with health status (p = 0.03 and 0.003, respectively). NFL and P-tau mean values demonstrated a significant difference between the ACS and control groups (p = 0.003 and 0.006), however Aß42 did not (p = 0.114). ROC analysis showed that plasma P-tau and NFL, with AUCs of 0.717 and 0.735, respectively, could substantially distinguish ACS from the HC group (p = 0.007 and 0.003, respectively). Both plasma P-tau (r = -0.389; p = 0.004) and NFL (r = -0.424; p = 0.001) levels were significantly and negatively correlated with individuals' MMSE scores. NFL and plasma P-tau show promise in differentiating AD patients from healthy individuals. However, similar larger studies are needed to validate our findings.

Therapeutic role of Rauwolfia serpentina in minimizing the risk of glycosylation and associated biomarkers in experimentally induced type 1 diabetic mice.

Present work investigates the effects of hydro-methanolic roots extract (HyMREt) of Rauwolfia serpentina in type 1 diabetic mice. Mice were divided into normal, diabetic, negative and positive controls (I-IV) and three test (HyMREt doses) groups (V-VII - 50, 100, &150mg/kg). Allocated treatment of each group was given orally for 14 days in overnight fasted state. Percent change in fasting blood glucose (FBG), body weights, body tissue weights, hepatic glycogen, total lipids, glycosylated hemoglobin (HbA1c), complete blood profile and antioxidant enzymes including catalase (CAT) and superoxide dismutase (SOD) were estimated. HyMREt doses produced meaningful (p<0.0001) reduction (-39 to -53%) in FBG. Hemoglobin (Hb) levels were raised, HbA1c were considerably decreased (4.5-3.77%) and glycosylation (HbA1c to Hb) ratio was expressively (p<0.0001) improved in test groups. Dose-wise improvement (p< 0.05) in total glycogen and decrement (p<0.05) in lipids were observed in livers of test groups. HyMREt significantly decreased (p<0.05) percent inhibition of SOD and CAT. HyMREt doses progressively (p<0.05) improved RBC and other hematological parameters while decrement was only noticed in leucocyte counts. Administration of test doses of HyMREt were significantly reduced the glycosylation, oxidative stress and anemia caused by alloxan intoxication in mice.

History in perspective: How Alzheimer's Disease came to be where it is?

Treatment of Alzheimer's Disease (AD) remains an unsolved issue despite the pronounced global attention it has received from researchers over the last four decades. Determining the primary cause of the disease is challenging due to its long prodromal phase and multifactorial etiology. Regardless, academic disagreements amongst the scientific community have helped in making significant advancements in underpinning the molecular basis of disease pathogenesis. Substantial development in fluid and imaging biomarkers for AD led to a sharp turn in defining the disease as a molecular construct, dispensing its clinical definition. With conceptual progress, revisions in the diagnostic criteria of AD were made, culminating into the research framework proposed by National Institute on Aging and Alzheimer's Association in 2018 which unified different stages of the disease continuum, giving a common language of AT(N)1 classification to researchers. With realization that dementia is the final stage of AD spectrum, its early diagnosis by means of cerebrospinal fluid biomarkers, Positron Emission Tomography and Magnetic Resonance Imaging of the brain holds crucial importance in discovering ways of halting the disease progression. This article maps the insights into the pathogenesis as well as the diagnostic criteria and tests for AD as these have evolved over time. A contextualized timeline of how the understanding of AD has matured with advancing knowledge allows future research to be directed and unexplored avenues to be prioritized.

Neonatal danger signs and healthcare seeking behaviours: A cross-sectional study in Karachi amongst pregnant females.

OBJECTIVE: To assess knowledge among pregnant women regarding neonatal danger signs and to evaluate their healthcare seeking behaviour. METHODS: The cross-sectional study was conducted at Civil Hospital, Karachi, from February to April 2018, and comprised pregnant females presenting to the hospital. They were interviewed through a structured questionnaire. Participants' knowledge regarding 12 neonatal danger signs was assessed. Those who could identify 3 or more such signs were considered to have a good knowledge. Data was analysed using SPSS 23. RESULTS: There were 468 subjects with a mean age of 25.17±5.18 years. Of the total, 416(88.9%) women were aged 18-35 years. Overall, 72(15%) subjects had good knowledge, while 366(78%) could identify at least one danger sign. Also, 318(82%) participants preferred visiting a hospital / clinic to seek healthcare. CONCLUSIONS: There was a severe lack of knowledge among pregnant women regarding neonatal danger signs. However, most participants realised the necessity of taking their neonate to a hospital / clinic for proper healthcare.

Knowledge and attitudes about burn complications in medical students.

Burn injury in developing countries is a major cause of morbidity and mortality. Pakistan faces a widespread problem of burn complications. The objective of this cross sectional study was to assess knowledge and attitudes in medical students regarding burn complications. A 12-question survey was distributed among 3rd, 4th and 5th year students of public sector medical colleges of Karachi, Pakistan. Out of 353 respondents, the proportion of students that were aware of burn complications was the following, with an expected response of 100% for each complication: depression [86%], fear [53.3%], post-traumatic stress disorder [40.8%], suicidal tendencies [35.1%], sleep disorder [30%], anxiety [27%], guilt [12.7%], personality changes [7%], eating disorder [5.4%], cardiac arrest [68%], hypothermia [64.3%], contractures [61%], infection [43.6%], chronic pain [41.6%], scarring [33%], chronic inflammation [24.9%], and muscle wasting [22.9%]. Most students were of the view that scarring occurs in third-degree burns [68%] only; being unaware of its presence in second-degree burns. 19.3% of the students incorrectly thought that surgery is needed for the healing of all burn patients. There appears to be a lack of basic knowledge about the physiological and psychological complications of burns among medical students of Pakistan.

Risk assessment of insulin resistance with anthropometric measures in non-diabetic heart failure patients: A gender based case-control investigation at tertiary care hospitals of Karachi, Pakistan.

To assess the risk of anthropometric measures and serum markers of insulin resistance in non-diabetic heart failure (NDHF) patients and the difference among male and female subjects. 53 males, 27 females NDHF patients were enrolled and 80 healthy subjects were matched as control. Anthropometric measures, fasting blood glucose level (FBGL) and serum insulin (SI) were measured. Insulin's function (ß-cells quantification) was computed through Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). Significant (p<0.05) difference in height was observed in NDHF male and female patients, whereas, weight was only significant in females. Higher mean values of FBGL, SI, and HOMA-IR in NDHF patients were observed. Highly significant (p<0.0001 & p<0.05) difference was also found in similar gender based case-control markers like FBGL, SI, and HOMA-IR. Phi coefficient for risk associations showed weak-positive correlation in both genders in FBGL, SI, and HOMA-IR. Anthropometric measures indicated lesser risk in both gender, especially in females' decreased risk in body height and waist circumference was observed. Risk measurement through odds ratios (OR) of FBGL and HOMA-IR in female subjects indicated significantly (p<0.001) double risk, whereas, in SI, three fold risks were measured in females as compared to male. Odds of exposure in cases were significantly (p<0.001) greater than in controls verified with significant Pearson and Fisher Exact Probability Test (FEPT) values with two-tailed estimates of probability in χ2 (Chi -Square) estimation test. Findings suggest increased risk of IR in female NDHF patients. Increased in FBGL IR and HOMA-IR and BMI were also found as distinguishing findings in NDHF cases.

Musculoskeletal pain and its associated risk factors among medical students of a public sector University in Karachi, Pakistan.

OBJECTIVE: To assess the frequency and anatomical distribution of musculoskeletal pain and its related risk factors. METHODS: This cross-sectional study was conducted at the Dow Medical College, Karachi, from September to November 2016, and comprised medical students. A self-administered questionnaire was filled out by the participants. Data was analysed using SPSS 17. RESULTS: Of the 360 participants, 268(74.4%)and 140(38.9%)students reported having musculoskeletal pain in at least one of the body sites in the past 12 months and in the past seven days, respectively. Self-reported mental stress (p=0.01) and history of trauma in the neck, shoulder or lower back (p=0.002) were the most significant predictors of musculoskeletal pain during the past seven days. Age, body mass index, gender, academic year, family history of musculoskeletal disorders, hours of clinical practice per day, hours of study per day, smoking and exercise had no significant relation with musculoskeletal pain in neck, shoulder or lower back (p>0.05 each). The use of computer/laptop for more than three hours per day had an increased occurrence of neck pain (p=0.03).. CONCLUSIONS: There was a significant risk of musculoskeletal pain for medical students.

Ameliorative effect of hydro-methanolic extract from roots of Rauwolfia serpentina on some biochemical parameters of type 1 diabetic mice.

Present work seeks to investigate the biochemical parameters in terms of hypoglycemic and hypolipidemic effects of hydro-methanolic roots extract (HyMREt) of Rauwolfia serpentina in type 1 (alloxan induced) diabetic mice. Animals were divided into seven groups, four control groups, and three were test groups (HyMREt at 50, 100, & 150mg/kg). Each treatment was repeated for 14 days regularly in all seven respective groups and afterwards the body weights, fasting blood glucose (FBG), insulin, and serum lipid levels were determined. Total body weights of diabetic mice treated with HyMREt extract were dose dependently (p<0.05) improved. FBG of test groups were significantly (p<0.0001) reduced in comparison with diabetic controls which displayed elevated fasting blood glucose level. The insulin levels of HyMREt treated groups were significantly (p<0.0001) higher than those of diabetic controls. Lower triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-c) and very low density lipoprotein cholesterol (VLDL-c) whereas elevated level of high density lipoprotein cholesterol (HDL-c) were observed in test dose treated groups. In comparison with diabetic controls, the converse levels of serum lipid were observed. Significant improvement in cardio-protective indices and coronary risk index was also observed. Findings of present study support the hypoglycemic and hypolipidemic potential of HyMREt of R. serpentina.

Homeostasis model assessment of insulin resistance in non-diabetic heart failure patients: A case-control investigation at tertiary care Hospitals of Karachi.

eart failure is a progressive, chronic disorder. Insulin resistance (IR) has been more and more involved as a preliminary metabolic perturbance predisposing to hyperglycemia, hyperlipidemia and atherosclerosis with others heart diseases. To investigate the relation of insulin resistance (IR) in non-diabetic heart failure patients this case-control study was carried out to ascertain the presence of IR with the aid of Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) in non-diabetic heart failure patients (NDHF patients) compared with healthy controls. The sample size was calculated for both, cases (NDHF patients) and control (healthy subjects), which was initially consisted of 113 respondents each. The study consisted of two phase duration. In Phase I, NDHF patients were approached initially; only 80 patients with NDHF completed the study procedure. In Phase II, 80 healthy subjects were targeted and matched. Fasting blood glucose level (FBGL) and serum insulin was estimated. Mathematical model to quantify ß-cell function and insulin resistance was also computed through Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) in both groups. Data was analyzed on SPSS version 16. Mean values with ± standard deviation (SD) of insulin (10.2±4.36) and HOMA-IR (2.52±1.15) were significantly (p<0.05) higher in NDHF patients as compared to control subject (6.4±3.39, 1.45±0.80). Average insulin to glucose ratio was 0.10±0.044 in NDHF patients which was significantly (p<0.0001) lowered in controls i.e., 0.073±0.039. Marginal and matrix plot analysis revealed that a higher patients count have had the HOMA-IR values <1.5 units while opposite scenario was observed in control group. Regression analyses of HOMA-IR with FBGL (as independent indicator) also authenticate the similar pattern. The present study concludes that insulin resistance (decreased insulin sensitivity) is a characteristic finding in Pakistani population of heart failure as compared to matched healthy controls.

Elevated fasting blood glucose is associated with increased risk of breast cancer: outcome of case-control study conducted in Karachi, Pakistan.

BACKGROUND: There are several validated risk factors for breast cancer. However the legitimacy of elevated fasting blood glucose (FBG) is not well established. This study was designed to assess this parameter as a risk factor for breast cancer among pre- and post-menopausal women. MATERIALS AND METHODS: This case-control study was conducted at Department of Biochemistry, University of Karachi from June 2010 to August 2014. Simple random sampling technique was used to collect data of study subjects comprising 175 diagnosed breast cancer patients with positive histopathology from Breast Clinic, surgical unit-1, Civil Hospital, Karachi and 175 healthy controls from various screening programs. Blood samples were analyzed for FBG and serum insulin. RESULTS: FBG, HOMA-IR, systolic and diastolic blood pressure were significantly raised in breast cancer cases when compared to control subjects. Cases and controls were further categorized in to two groups using cutoff value of 110 mg/dl to distinguish subjects into normal fasting glucose (<110 mg/dl) and having impaired fasting glucose (≥110-≤125 mg/dl) or diabetes (≥126 mg/dl). Odds ratios were found to be 1.57, 2.15 and 1.17 in overall, pre-menopausal and post-menopausal groups, respectively. (all p<0.05). CONCLUSIONS: A statistically significant risk of breast cancer exists in women having elevated fasting blood glucose levels, corresponding to prediabetes and diabetes, among pre and postmenopausal ages, with comparatively greater effects in the premenopausal group.