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.

Cancer Stem Cells and Treatment of Cancer: An Update and Future Perspectives.

Cancer stem cells (CSCs) play an essential role in tumour progression and metastasis. These cells have the unique ability to self-renew and differentiate into specific tissue cell types. Their capacity for self-renewal enables CSCs to persist over time, thereby contributing to cancer relapse and therapy resistance. Therefore, targeting CSCs has emerged as a promising cancer treatment strategy. CSCs exhibit differentiation, self-renewal, and plasticity, and they contribute to multiple aspects of malignant tumours, such as recurrence, metastasis, heterogeneity, multidrug resistance, and radiation resistance. While conventional treatments predominantly target cancer cells that are not CSCs, CSCs frequently survive, resulting in tumour recurrence and relapse. This article concentrates on the development of novel therapeutic strategies that combine conventional treatments with CSC inhibitors to eradicate cancer cells and CSCs, thereby treating cancer and preventing its recurrence. However, the diversity of CSCs poses a significant obstacle to the development of CSC-targeted therapies, necessitating extensive research for a better understanding and exploration of therapeutic approaches. Future development of CSC-targeted therapies will rely heavily on overcoming this obstacle.

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.

Understanding COVID-19 vaccine acceptance in Pakistan: an echo of previous immunizations or prospect of change?

BACKGROUND: As countries ramp up their COVID-19 vaccination programs, attitudes of the population remain a determining player in the success of these plans. This study analyses the factors associated with intent to vaccinate against COVID-19 in the Pakistani population. METHODOLOGY: This cross-sectional, anonymous, online survey was carried out in April 2021. Participants' demographic details, experiences relating to COVID-19 and its vaccination, and their health beliefs were inquired and divided across Health Belief Model constructs. Multivariable regression was used to determine factors associated with a No/Not sure vs Yes response for vaccination intention. RESULTS: Of the 655 respondents, 62.0% were willing to get vaccinated. Significant predictors of a less likelihood of resisting vaccination included advanced age (AOR 0.25; 95% CI 0.07-0.88), fear of contracting COVID-19 (AOR 0.47; 95% CI 0.27-0.82), hope of preventing its spread (AOR 0.30; 95% CI 0.19-0.49), and community pressure (AOR 0.22; 95% CI 0.13-0.37). Concerns about vaccine reliability (AOR 2.75; 95% CI 1.67-4.53) and religious inhibitions (AOR 2.45; 95% CI 1.34-4.48) swayed people away from vaccination. CONCLUSION: Despite a reasonably good response of Pakistanis to vaccination, factors negatively influencing their intention need to be timely addressed to control this pandemic.

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.