AATF inhibition exerts antiangiogenic effects against human hepatocellular carcinoma.

Background and aims: Angiogenesis is a key factor in the growth and metastasis of hepatic tumors and thus a potential therapeutic target in hepatocellular carcinoma (HCC). In this study, we aim to identify the key role of apoptosis antagonizing transcription factor (AATF) in tumor angiogenesis and its underlying mechanisms in HCC. Methods: HCC tissues were analyzed for AATF expression by qRT-PCR and immunohistochemistry. Stable clones of control and AATF knockdown (KD) were established in human HCC cells. The effect of AATF inhibition on the angiogenic processes was determined by proliferation, invasion, migration, chick chorioallantoic membrane (CAM) assay, zymography, and immunoblotting techniques. Results: We identified high levels of AATF in human HCC tissues compared to adjacent normal liver tissues, and the expression was found to be correlated with the stages and tumor grades of HCC. Inhibiting AATF in QGY-7703 cells resulted in higher levels of pigment epithelium-derived factor (PEDF) than controls due to decreased matric metalloproteinase activity. Conditioned media from AATF KD cells inhibited the proliferation, migration, and invasion of human umbilical vein endothelial cells as well as the vascularization of the chick chorioallantoic membrane. Furthermore, the VEGF-mediated downstream signaling pathway responsible for endothelial cell survival and vascular permeability, cell proliferation, and migration favoring angiogenesis was suppressed by AATF inhibition. Notably, PEDF inhibition effectively reversed the anti-angiogenic effect of AATF KD. Conclusion: Our study reports the first evidence that the therapeutic strategy based on the inhibition of AATF to disrupt tumor angiogenesis may serve as a promising approach for HCC treatment.

Bioinformatic Tools for the Identification of MicroRNAs Regulating the Transcription Factors in Patients with ß-Thalassemia.

ß-thalassemia is a significant health issue worldwide, with approximately 7% of the world's population having defective hemoglobin genes. MicroRNAs (miRNAs) are short noncoding RNAs regulating gene expression at the post-transcriptional level by targeting multiple gene transcripts. The levels of fetal hemoglobin (HbF) can be increased by regulating the expression of the γ-globin gene using the suppressive effects of miRNAs on several transcription factors such as MYB, BCL11A, GATA1, and KLF. An early step in discovering miRNA:mRNA target interactions is the computational prediction of miRNA targets that can be later validated with wet-lab investigations. This review highlights some commonly employed computational tools such as miRBase, Target scan, DIANA-microT-CDS, miRwalk, miRDB, and micro-TarBase that can be used to predict miRNA targets. Upon comparing the miRNA target prediction tools, 4 main aspects of the miRNA:mRNA target interaction are shown to include a few common features on which most target prediction is based: conservation sites, seed match, free energy, and site accessibility. Understanding these prediction tools' usage will help users select the appropriate tool and interpret the results accurately. This review will, therefore, be helpful to peers to quickly choose a list of the best miRNAs associated with HbF induction. Researchers will obtain significant results using these bioinformatics tools to establish a new important concept in managing ß-thalassemia and delivering therapeutic strategies for improving their quality of life.

COVID-19 Effects on Geriatric Population and Failures of Aminoquinoline Therapy: Compilation of Studies from EU, USA, and China; Safety and Efficacy of Vaccines in the Prevention and Treatment of COVID-19.

BACKGROUND: Mortality of the older adult population suffering from COVID-19 has been increasing at an alarming rate, and people older than 76 years of age reported 18% mortality. Mainly, the EU and USA exhibited a greater fatality rate due to lack of selective immunization and anti-SARS Co-V-2 therapeutics. Very limited reports are available to delineate the impact of COVID-19 on the geriatric population and the failures of aminoquinoline therapy. METHODS: We performed a substantial literature review in the PubMed/Medline databases to extract the information pertaining to the COVID-19 impact on the geriatric population and recent failures of aminoquinoline therapy in COVID-19 patients of EU, China, USA and the requirement of vaccination. RESULTS AND DISCUSSIONS: Both parental strains and mutant variants of SARS Co-V-2 can induce severe respiratory complications, multiorgan failure, and clotting abnormalities in older adults due to low immunocompetence. Aminoquinoline derivatives, such as chloroquine (CQ) and hydroxychloroquine (HCQ), are preferred primarily for COVID-19 treatment, but several controversies are being reported for its usage worldwide. In this review, we have provided the effects of COVID-19 on the geriatric population of EU and an overview of the mechanism of action of aminoquinolines. Furthermore, CQ and HCQ are not the preferred choice of drugs if the COVID-19 patients already have existing co-morbid conditions viz., diabetes mellitus and hypertension. CONCLUSION: A new advent of COVID-19 vaccines, such as nucleic acid-based (DNA/mRNA) vaccines, protein subunit vaccines, viral vector vaccines, and inactivated vaccines, have been developed for treating SARS-CoV-2 infection after the failure of aminoquinoline therapy in EU, China, and USA patients. However, some of the vaccines are yet to be examined against mutant strains of SARS CoV-2 that originated in the UK, Nigeria, South Africa, Brazil, and India.

Role of NRF2 cascade in determining the differential response of cervical cancer cells to anticancer drugs: an in vitro study.

BACKGROUND: Cervical cancers are usually treatable if detected in early stages by a combination of therapies. However, the prognosis of cervical cancer patients with metastasis remains unfavorable due to the fact that most of the cervical carcinomas are either resistant to anticancer drugs or show signs of relapse after initial treatment. Therefore, it is important to control the chemoresistance as it is the key to develop effective treatment options for cervical cancer. OBJECTIVE: The current study aimed at evaluating the differential responses of cervical cancer cells to anti-cancer drugs and assessed whether the differences in the expression profiles of antioxidant genes regulated by nuclear factor erythroid-2-related factor 2 (NRF2), led to the variations in the sensitivities of the cancer cells to treatment. METHODOLOGY: Three cervical cancer cell lines were investigated for their differences in NRF2 pathway by measuring the gene expression and enzyme activity. The differences in the sensitivity to anti-cancer drugs and variation in ROS profile was also evaluated. The addition of exogenous drugs to manipulate the intracellular ROS and its effect on NRF2 pathway genes was also investigated. RESULTS: HeLa and SiHa cells were more sensitive to cisplatin and oxaliplatin treatment than C33A cells. HeLa and SiHa cells had significantly lower NRF2 gene levels, NQO1 enzyme activity and basal GSH levels than C33A cells. Levels of ROS induced were higher in HeLa than C33A cells. CONCLUSION: Overall, the differences in the cellular levels of antioxidant regulatory genes led to the differential response of cervical cancer cells to anti-cancer drugs.

A Review on SARS-CoV-2-Induced Neuroinflammation, Neurodevelopmental Complications, and Recent Updates on the Vaccine Development.

Coronavirus disease 2019 (COVID-19) is a devastating viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The incidence and mortality of COVID-19 patients have been increasing at an alarming rate. The mortality is much higher in older individuals, especially the ones suffering from respiratory distress, cardiac abnormalities, renal diseases, diabetes, and hypertension. Existing evidence demonstrated that SARS-CoV-2 makes its entry into human cells through angiotensin-converting enzyme 2 (ACE-2) followed by the uptake of virions through cathepsin L or transmembrane protease serine 2 (TMPRSS2). SARS-CoV-2-mediated abnormalities in particular cardiovascular and neurological ones and the damaged coagulation systems require extensive research to develop better therapeutic modalities. As SARS-CoV-2 uses its S-protein to enter into the host cells of several organs, the S-protein of the virus is considered as the ideal target to develop a potential vaccine. In this review, we have attempted to highlight the landmark discoveries that lead to the development of various vaccines that are currently under different stages of clinical progression. Besides, a brief account of various drug candidates that are being tested to mitigate the burden of COVID-19 was also covered. Further, in a dedicated section, the impact of SARS-CoV-2 infection on neuronal inflammation and neuronal disorders was discussed. In summary, it is expected that the content covered in this article help to understand the pathophysiology of COVID-19 and the impact on neuronal complications induced by SARS-CoV-2 infection while providing an update on the vaccine development.

Impact of experiential learning among medical undergraduates: A randomized controlled trial.

BACKGROUND: Experiential learning sessions as a teaching aid have been applied early in the medical undergraduate curriculum to improve the knowledge and inculcate research interest. We compared the ability of 1st-year medical undergraduates to answer the molecular biology questions among those who had attended the experiential learning sessions of molecular biology techniques versus those who did not attend. SUBJECTS AND METHODS: A randomized controlled trial was carried out with 200 1st-year medical undergraduates, among whom 69 students were selected by simple random sampling for the demonstration of the molecular biology techniques, such as isolation of genomic DNA, polymerase chain reaction, cell culture techniques, western blotting, and high-performance liquid chromatography for 1-week duration. Student's feedback was collected on a five-point Likert sc ale at the end of the session to understand how they agree or disagree with a particular statement. The content validity rate (CVR) and content validity index (CVI) of the questionnaire were determined, and its internal consistency was examined by Cronbach's alpha. The internal assessment marks of these students, valued by faculty who were blinded to their training sessions, were compared with the rest of the 131 students by independent t-test to know the outcome of these experiential learning sessions. RESULTS: On CVR and CVI assessment, all the questions scored more than 0.70 and 0.85, respectively. Cronbach's alpha for the whole questionnaire was 0.85. Student's feedback indicated that these sessions did complement the cognitive skills acquired for these techniques. We also found a statistically significant improvement (P = 0.006) in the examination performance between the students who attended versus those who did not attend the experiential learning sessions. CONCLUSION: Experiential learning, through demonstration and hands-on experience, enhance d the learning of molecular biology techniques among 1st-year medical undergraduates.

Major Protein of Carcinoembryonic Antigen Gene Family - CD66c, A Novel Marker in Colon Carcinoma.

INTRODUCTION: In view of rising trend of the incidence of colorectal carcinoma in the Indian population due to adoption of western lifestyles and behaviours, we investigated the expression of the new emerging stem cell biomarker, CD66c in colorectal carcinoma of Indian origin. AIM: To study the expression of CD66c in human colorectal carcinoma and to correlate level of marker expression with tumour staging. MATERIALS AND METHODS: This hospital based prospective study was conducted on 26 colorectal carcinoma patients in the age group of 20 years to 70 years. Surgically resected tumour specimens along with adjacent normal tissue were collected taking necessary precautions, paraffin embedded sections were prepared and used for histological and immunohistochemical analysis of CD66c. STATISTICAL ANALYSIS: Descriptive statistical measures like mean, standard deviation, percentage was applied. Other inferential statistical tests like Chi-square, Fisher's-exact test and one-way ANOVA was applied to find out the association of CD66c with different stages. The difference were interpreted as statistically significant when p <0.05. RESULTS: CD66c showed differential expression with membrane positivity in normal colorectal epithelial cells and cytoplasmic expression in tumour cells. There was significant correlation between CD66c expression and tumour site (p=0.02) with colon carcinoma showing positive expression compared to the rectal carcinoma. There was no significant correlation between CD66c staining and tumour stage (p=0.947). No significant relationship was observed between CD66c expression and other clinicopathologic variables studied such as sex (p=0.552), age (p=0.713) and tumour grade (p=0.263). CONCLUSION: CD66c can be specifically used for colon carcinoma and may be a novel marker in colon carcinoma stem cell isolation. The quantification of CD66c can be further verified by flow cytometry and RT-PCR. Further studies can be carried out using CD66c alone or in combination with other markers to develop cancer stem cell directed therapy.

Comparative assessment of cytokines and other inflammatory markers for the early diagnosis of neonatal sepsis-a case control study.

OBJECTIVE: Cytokines (IL-6, IL-8 and TNF-α), sCD163, and C-reactive protein were serially measured in an attempt to identify a set of tests which can reliably confirm or refute the diagnosis of neonatal sepsis at an early stage. METHODS: One hundred neonates suspected to have sepsis on clinical grounds and who met the inclusion criteria were enrolled for the study. Based on the positive or negative blood culture reports they were classified as infected (n=50) and non-infected (n=50) neonates respectively. Fifty healthy neonates without any signs of sepsis were also included in the study as control group. The initial blood sample was taken on day 0 (at the time of sepsis evaluation) and two further samples were taken on days 1 and 2 for monitoring the clinical progress and response to treatment. In the control group the cord blood and 48 hours venous sample was collected. Plasma CRP (ng/ml), IL-6 (pg/ml), IL-8 (pg/ml), TNF-α (ng/ml) and sCD163 (ng/ml) were determined by double antibody method Enzyme Linked Immunosorbent Assay in all the three blood samples. RESULTS: The cut of levels for CRP at >19,689 ng/ml had a sensitivity of 68%, specificity of 92%, for IL-6 at >95.32 pg/ml had a sensitivity of 54%, specificity of 96%, for IL-8 at >70.86 pg/ml had a sensitivity of 78%, specificity of 70%, for sCD163 at >896.78 ng/ml had a sensitivity of 100%, specificity of 88% for the diagnosis of infection before antibiotics. TNF-α levels of >12.6 ng/ml showed 100% sensitivity and 72% specificity for the diagnosis of inflammation. CONCLUSION: The most powerful predictor to differentiate between the non-infected and infected neonates before antibiotics was sCD163. The most powerful indicator for evaluation of prognosis is IL-6. sCD163 can be used alone to screen for sepsis in neonates before the results of blood culture are received.

Can soluble CD163 predict outcome of patients with acute respiratory distress from mechanical ventilation?: A pilot study.

PURPOSE: CD163 is a monocyte/macrophage-associated antigen which has recently been identified as a hemoglobin scavenger receptor and has also anti-inflammatory properties and an immunoregulatory role. This surface receptor undergoes ectodomain shedding upon an inflammatory stimulus, leading to increased fraction of soluble CD163 (sCD163). Hence, we hypothesized that the mechanical ventilation (MV) which is known to elicit inflammatory response may cause increased serum levels of sCD163 which can predict the outcome of patients from MV. SUBJECTS AND METHODS: Thirty patients with acute respiratory distress aged >18 years who required MV were enrolled for the study. Serum levels of sCD163 were estimated using quantitative immunometric sandwich enzyme immunoassay technique from 3 mL of the venous blood sample which was collected immediately and at 24 h after the patient was connected to MV. On the basis of the outcome of the patient from MV, they were divided into two groups; survivors and nonsurvivors. RESULTS: Out of the 30 patients, 18 patients survived and 12 patients expired. Serum levels of sCD163 were significantly increased in nonsurvivors when compared with survivors (P < 0.01) at 24 h after connecting to MV. sCD163 > 1020 ng/mL at 24 h of MV increases the probability of mortality by factor 6. An increase of sCD163 by 1 ng/mL significantly increases the relative probability of mortality by a factor of 1.0017 (95% confidence interval, 1.0004-1.0030, P = 0.0005). CONCLUSIONS: Elevated levels of sCD163 at 24 h of MV help in predicting the outcome of patients with acute respiratory failure from MV.