Evaluation of molecular effects associated with apoptosis, tumour progression, angiogenesis and metastasis by a novel combination of drugs with ormeloxifene in triple negative breast cancer cells.

Aim: To investigate the molecular effects of a novel combination [sertraline and plumbagin (comb) with ormeloxifene (Orm)] for anticancer activity in triple negative breast cancer cell line "MDA-MB-231". Methods: The cytotoxic effect of the drugs was analyzed by the MTT assay and nuclear morphological changes by acridine orange/ethidium bromide (AO/EB) staining. Induction of apoptosis by annexin V-FITC staining, active caspase-3 detection and cell cycle analysis were studied in vitro on "MDA-MB-231" cells. The qRT-PCR was done to explore the upregulation and down regulation of targeted genes for angiogenesis, metastasis, tumor suppression and protein folding on the triple negative breast cancer cells. The preliminary anti-angiogenic effect of the drugs was assessed by chorioallantoic membrane (CAM) assay. Results: Orm showed inhibitory effects in "MDA-MB-231" cells in a dose and time dependent manner whereas; the drugs in combination gave better cytotoxic effects in the screening MTT assay. Orm + comb was more effective than Orm alone in eliciting apoptosis as well as inhibited the single cell to grow into a colony. CAM assay using Orm and Orm + comb suggested the anti-angiogenic potential which was further confirmed by the downregulation of VEGF in "MDA-MB-231" cells by qRT-PCR studies. The combination was found to effectively upregulate the expression of P53 and P21 and downregulate the gene expression of zinc finger E-box binding homeobox 1 (ZEB1) and heat shock protein 70 (HSP70) in "MDA-MB-231" cancer cells. Conclusions: Collectively this study reveals the efficacy of Orm + comb as more significant than the clinically used tamoxifen (Tam). The study elucidates the promising novelty of the combination as a potential chemotherapeutic intervention for mitigating the aggressiveness of triple negative breast cancer and it addresses the intrinsic resistance caused by single drug treatments.

A hybrid electrocoagulation-biocomposite adsorption system for the decolourization of dye wastewater.

Among the various methods for the removal of azo dye, electrocoagulation is recognized to be highly efficient. However, the process is associated with high operation and maintenance cost, which demands the need for reducing the electrolysis time without compromising the performance efficiency. This can be achieved by adopting hybrid electrocoagulation process with a low-cost but effective process, such as adsorption. The study investigated the performance of a hybrid electrocoagulation-biocomposite system (H-EC-BC) for removing methyl orange dye. Firstly, the operating parameters of electrocoagulation process were optimized and a removal efficiency of 99% has been attained using Fe-SS electrodes at a pH of 6 for a reaction time of 30 min. The performance of EC process was found to be decreasing with increase in dye concentration. Secondly, biocomposite was synthesized from Psidium guajava leaves and characterized using SEM, FTIR, EDAX, and XRD analyses. The results suggested that it is having a porous nature and cellulose crystal structure and confirmed the presence of chemical elements such as carbon (65.2%), oxygen (29.1%) as primary with Fe, Cl, Na and Ca as secondary elements. The performance of the biocomposite was evaluated for the dye adsorption using spectrophotometric methods. Various operating parameters were optimized using experimental methods and a maximum removal efficiency of 65% was achieved at a pH of 6, dosage of 5 g/L and an adsorption contact time of 120 min. The maximum efficiency (92.78%) was obtained with Fe-SS electrodes and KCl as a sustaining electrolyte under acidic circumstances (pH 6). The biocomposite was observed to be more efficient for higher dye concentration. Langmuir and Freundlich adsorption isotherms were fitted with the experimental results with R2 values as 0.926 and 0.980 respectively. The adsorption kinetics were described using Pseudo-first and Pseudo-second order models, wherein Pseudo-second order model fits the experimental results with R2 value of 0.999. The energy consumption of electrocoagulation (EC) process in the hybrid H-EC-BC system was compared to that of a standard EC process. The results demonstrated that the hybrid system is approximately 7 times more energy efficient than the conventional process, thereby implicating its adaptability for field application.

Spinal muscular atrophy: Molecular mechanism of pathogenesis, diagnosis, therapeutics, and clinical trials in the Indian context.

Spinal muscular atrophy (SMA) is a neuromuscular, rare genetic disorder caused due to loss-of-function mutations in the survival motor neuron-1 (SMN1) gene, leading to deficiency of the SMN protein. The severity of the disease phenotype is inversely proportional to the copy number of another gene, SMN2, that differs from SMN1 by a few nucleotides. The current diagnostic methods for SMA include symptom-based diagnosis, biochemical methods like detection of serum creatine kinase, and molecular detection of disease-causing mutations using polymerase chain reaction (PCR), multiplex ligation-dependent probe amplification (MLPA), and exome or next-generation sequencing (NGS). Along with detection of the disease-causing mutation in the SMN1 gene, it is crucial to identify the copy number of the SMN2 gene, which is a disease modifier. Therapeutic options like gene therapy, antisense therapy, and small molecules are available for SMA, but, the costs are prohibitively high. This review discusses the prevalence, diagnosis, available therapeutic options for SMA, and their clinical trials in the Indian context, and highlights the need for measures to make indigenous diagnostic and therapeutic interventions.

Candida utilis: a rare cause of septicemia in children.

Candida utilis is an emerging fungal pathogen in blood. The main aim of this study was to describe the prevalence, methods of speciation and antifungal susceptibility of Candida utilis at a tertiary care centre. METHODS: This was a retrospective study carried out at a tertiary care centre in South India. Over a period of 1 year, three Candida utilis were isolated from blood culture identified by MALDI-TOF MS Version 3.2 and were confirmed by ITS sequencing. Susceptibility testing was carried out by micro broth dilution. RESULTS: All three patients had a common risk factor of prolonged ICU stay but the source of infection could not be identified. Candida utilis isolates were identified by MALDI-TOF and confirmed by ITS sequencing. They were pansusceptible to all tested antifungal drugs. Among these, two patients who were treated in hospital had good clinical outcome and response to antifungal drugs. A third patient was lost to follow up. CONCLUSION: Candida utilis was predominantly seen between 0-3 month olds. Conventional methods of speciation were unable to identify C. utilis to species level. Rapid identification was done by MALDI-TOF MS and confirmed by sequencing. Rapid identification leads to prompt treatment and favours a good clinical outcome.

Variable reactivity of Rh D antigen and its serological characterization.

Background: Variation in the reactivity on Rh D typing may pose challenges in interpretation and ambiguity in further patient management.Materials and Methods: A prospective study was conducted in the department of transfusion medicine for a period of 18 months. Blood grouping was performed by fully automated equipment employing column agglutination technique. All the samples with Rh D negative or discrepant reactions were subjected to weak D testing by the antihuman globulin testing method. Samples that tested positive were categorized as serological weak D type or Variant D and were further phenotyped with Partial D typing set with 6 monoclonal anti D antisera.Results: A total of 82,824 samples were tested for Rh D type during the study period. Of the study population, 65.7% were males. On Rh D type majority were Rh D positive (93%), 6.9% were negative, and the result was discrepant in 0.1% (70) samples. The overall prevalence of variant D was 1.28% (75) of the Rh D negative population and 0.09% of the total study population. The detection rate of variant D phenotype was significantly higher by the Column agglutination technique. Upon testing with Partial D kit, the partial D variant in the majority reacted wil all the 6 antisera and hence we could not rule out DIII(60%), in rest it was inconclusive. In 43% of subjects with Rh D discrepancy 'C' antigen was found in a homozygous state.Conclusion: The introduction of partial D typing kit alone may not help in the absolute characterization of variant D. Extended serological testing and selective integration of molecular testing is the need of the hour.