Studies on the hepatic and renal status of patients with sickle cell disease from western zone of Maharashtra, India

Background: Sickle cell disease (SCD) is the most common inherited monogenic genetic disorder in Indian tribal and non-tribal population. This condition is caused by mutations in the hemoglobin gene and inherited in an autosomal recessive pattern. Pathogenesis in SCD varies widely from patient to patient. Most of the infections affect SCD pathogenesis, so early diagnosis of the same is important.Methods: The present study was designed to evaluate the biochemical parameters to assess the hepatic and renal status in SCD subjects from west zone of Maharashtra, India. Patients with sickle cell disease (n=50) from primary health centres of Palghar were included in this study and age and sex matched healthy persons (n=50) were controls. Informed written consent was obtained from all the study subjects.Results: Our findings showed that Aspartate trasaminase (AST), Alanine transaminase (ALT), bilirubin and creatinine increased significantly above normal level in SCD subjects. Albumin and urea levels in SCD were found to have decreased in the SCD subjects. There is a slight increase in uric acid and creatinine levels; this indicates an adverse effect on hepatic function and moderate effect on renal function in sickle cell anemia patients. Most common events of SCD pathogenesis, can be categorized into hemolytic events and vaso-occlusive crisis-based events. Adverse effect on hepatic function can lead to further hemolytic events.Conclusions: Although specific biomarkers related to these different events needs to understand for assessment of pathogenesis, the ones we have studied can be useful to assess the status of hepatic and renal function to follow the effectiveness of therapeutic interventions.

Modulation of topoisomerase IIα expression and chemosensitivity through targeted inhibition of NF-Y:DNA binding by a diamino p-anisyl-benzimidazole (Hx) polyamide

BACKGROUND: Sequence specific polyamide HxIP 1, targeted to the inverted CCAAT Box 2 (ICB2) on the topoisomerase IIα (topo IIα) promoter can inhibit NF-Y binding, re-induce gene expression and increase sensitivity to etoposide. To enhance biological activity, diamino-containing derivatives (HxI*P 2 and HxIP* 3) were synthesised incorporating an alkyl amino group at the N1-heterocyclic position of the imidazole/pyrrole. METHODS: DNase I footprinting was used to evaluate DNA binding of the diamino Hx-polyamides, and their ability to disrupt the NF-Y:ICB2 interaction assessed using EMSAs. Topo IIα mRNA (RT-PCR) and protein (Immunoblotting) levels were measured following 18h polyamide treatment of confluent A549 cells. γH2AX was used as a marker for etoposide-induced DNA damage after pre-treatment with HxIP* 3 and cell viability was measured using Cell-Titer Glo®. RESULTS: Introduction of the N1-alkyl amino group reduced selectivity for the target sequence 5'-TACGAT-3' on the topo IIα promoter, but increased DNA binding affinity. Confocal microscopy revealed both fluorescent diamino polyamides localised in the nucleus, yet HxI*P 2 was unable to disrupt the NF-Y:ICB2 interaction and showed no effect against the downregulation of topo IIα. In contrast, inhibition of NF-Y binding by HxIP* 3 stimulated dose-dependent (0.1-2μM) re-induction of topo IIα and potentiated cytotoxicity of topo II poisons by enhancing DNA damage. CONCLUSIONS: Polyamide functionalisation at the N1-position offers a design strategy to improve drug-like properties. Dicationic HxIP* 3 increased topo IIα expression and chemosensitivity to topo II-targeting agents. GENERAL SIGNIFICANCE: Pharmacological modulation of topo IIα expression has the potential to enhance cellular sensitivity to clinically-used anticancer therapeutics. This article is part of a Special Issue entitled: Nuclear Factor Y in Development and Disease, edited by Prof. Roberto Mantovani

The Usability of E-learning Platforms in Higher Education: A Systematic Mapping Study

The use of e-learning in higher education has increased significantly in recent years, which has led to several studies being conducted to investigate the usability of the platforms that support it. A variety of different usability evaluation methods and attributes have been used, and it has therefore become important to start reviewing this work in a systematic way to determine how the field has developed in the last 15 years. This paper describes a systematic mapping study that performed searches on five electronic libraries to identify usability issues and methods that have been used to evaluate e-learning platforms. Sixty-one papers were selected and analysed, with the majority of studies using a simple research design reliant on questionnaires. The usability attributes measured were mostly related to effectiveness, satisfaction, efficiency, and perceived ease of use. Furthermore, several research gaps have been identified and recommendations have been made for further work in the area of the usability of online learning

Studies on the hepatic and renal status of patients with sickle cell disease from western zone of Maharashtra, India

Background: Sickle cell disease (SCD) is the most common inherited monogenic genetic disorder in Indian tribal and non-tribal population. This condition is caused by mutations in the hemoglobin gene and inherited in an autosomal recessive pattern. Pathogenesis in SCD varies widely from patient to patient. Most of the infections affect SCD pathogenesis, so early diagnosis of the same is important.Methods: The present study was designed to evaluate the biochemical parameters to assess the hepatic and renal status in SCD subjects from west zone of Maharashtra, India. Patients with sickle cell disease (n=50) from primary health centres of Palghar were included in this study and age and sex matched healthy persons (n=50) were controls. Informed written consent was obtained from all the study subjects.Results: Our findings showed that Aspartate trasaminase (AST), Alanine transaminase (ALT), bilirubin and creatinine increased significantly above normal level in SCD subjects. Albumin and urea levels in SCD were found to have decreased in the SCD subjects. There is a slight increase in uric acid and creatinine levels; this indicates an adverse effect on hepatic function and moderate effect on renal function in sickle cell anemia patients. Most common events of SCD pathogenesis, can be categorized into hemolytic events and vaso-occlusive crisis-based events. Adverse effect on hepatic function can lead to further hemolytic events.Conclusions: Although specific biomarkers related to these different events needs to understand for assessment of pathogenesis, the ones we have studied can be useful to assess the status of hepatic and renal function to follow the effectiveness of therapeutic interventions.

DNA-binding properties of new fluorescent AzaHx-amides: Methoxy-pyridyl-aza-benzimidazole-pyrrole-imidazole/pyrrole

DNA minor groove binding polyamides have been extensively developed to control abnormal gene expression. Establishment of the novel, inherently fluorescent Hx-amides has provided an alternative path for studying DNA binding in cells by direct observation of cell localization. Because of the 2:1 antiparallel stacking homodimer binding mode of these molecules to DNA, modification of Hx-amides to AzaHx-amides has successfully extended the DNA recognition repertoire, from central CG (recognized by Hx-I) to central GC (recognized by AzaHx-P) recognition. To potentially target two consecutive GG bases, new modifications from AzaHx moiety to 3-Pyr-AzaHx and 2-Pyr-AzaHx moieties were developed. The newly designed molecules are also small-sized, fluorescent amides with Pyr-AzaHx connected to two conventional five-membered heterocycles. Complementary biophysical methods were carried out to investigate DNA binding properties of these molecules. The results showed that neither 3-Pyr-AzaHx nor 2-Pyr-AzaHx was able to mimic I-I to specifically target GG dinucleotides. Rather, 3-Pyr-AzaHx functions like AzaHx or f-I or P-I as a antiparallel stacked dimer. 3-Pyr-AzaHx-PI (2) binds 5'-ACGCGT'-3' with improved binding affinity and high sequence specificity when compared to its parent molecule AzaHx-PI (1). However, 2-Pyr-AzaHx is detrimental to DNA binding because of an unfavorable steric clash upon stacking in the minor groove