Developmental toxicity and neurobehavioral effects of sodium selenite and selenium nanoparticles on zebrafish embryos.

Selenium, a trace mineral, is essential for several physiological processes in humans and animals. It is an antioxidant vital for the immunological response, DNA synthesis, thyroid hormone metabolism, and antioxidant defense enzymes. Zebrafish embryos and larvae were exposed to different concentrations of sodium selenite (SodSe) and selenium nanoparticles (SeNs) at various developmental stages. The study evaluated the impact of SodSe and SeNs on larvae survival, hatching rate, and morphological abnormalities. Also, acridine orange staining was used to analyze the apoptotic cell death, and behavioral tests were conducted to assess anxiety-like behaviors. The results showed that both SodSe and SeNs influence the development and neurobehavior of zebrafish larvae in a concentration-dependent manner. SodSe at high concentration causes low survival rates, delayed hatching, and increased morphological defects in zebrafish larvae. In addition, exposure to SodSe resulted in elevated apoptosis in different larval tissues. Zebrafish larvae treated with SodSe and SeNs exhibited anxiety-like behaviour, increased thigmotaxis, less exploratory behaviour, and less swimming patterns. The nerve conductions and stimuli responses evaluated through acetylcholine esterase (AChE) and cortisol assays, revealed a decrease in the activity in a dose-dependent manner of SodSe and SeNs. Interestingly, the effects of SeNs were lower even at higher concentrations when compared with SodSe at lower concentrations on zebrafish embryos. This shows that SeNs synthesized through biological methods may be less toxic and may have lower effect on the development and neurobehavior of zebrafish larvae. Thus, our study confirms the cytotoxic and neurobehavioral effects of SodSe and suggests the use of SeNs at lower concentration to provide insights into better understanding of developmental stages and metabolic pathways in zebrafish larvae.

Exploring the mitochondrial genome of Caridina pseudogracilirostris: a comparative analysis within the Atyidae Family.

BACKGROUND: Caridina pseudogracilirostris is a highly adaptive estuarine species found in brackish waters and marshes along the southwestern and southern coastal regions of India. METHODS AND RESULTS: The whole mitochondrial genome of C. pseudogracilirostris is 15,451 bp in length with 59.3% AT content and encodes 37 genes, including 22 tRNAs, 13 protein-coding genes, and two rRNAs, which are arranged in a distinctive pattern similar to most crustaceans. ML and BI methods were used for phylogenetic analysis of C. pseudogracilirostris clustered with other Caridina species, supporting the monophyly of the Caridina genus within the Atyidae family. The fully annotated mitochondrial genome of C. pseudogracilirostris was submitted to GenBank under accession number OQ534868.1. CONCLUSIONS: We are the first to report on the C. pseudogracilirostris whole mitochondrial genome, which provides a valuable resource for future research on genetics, evolution, phylogenetics, etc., among Caridina species and other species. The phylogenetic investigation supports the monophyly of the Caridina genus within the Atyidae family and emphasizes the value of mitochondrial genome data in determining the evolutionary relationships among crustaceans.

Real-time imaging and developmental biochemistry analysis during embryogenesis of Caridina pseudogracilirostris.

This paper reports on the real-time imaging and developmental biochemistry of the freshwater caridean shrimp, Caridina pseudogracilirostris. The complete time-lapse development of a single embryo was recorded in an artificial mold, developed in our lab, and imaged under a stereomicroscope. It took 8 days to complete the 5 stages of embryonic development (1 cleavage stage, 2 gastrulation stage, 3 nauplius stage, 4 prehatching embryo, and 5 zoea stages). As the decapod eggs are enriched with dense yolk, biochemical determination of the major components was made to evaluate the yolk utilization during embryogenesis. The concentration of protein, lipid, and carbohydrate declined drastically from Stage I (cleavage) to Stage IV (Zoea), reflecting sustained yolk utilization during embryogenesis. The increase in the size of the embryo correlates with changes in water content. Lipids, being the principal organic substrate, changes in the fatty acid (FA) composition of embryos during development were determined by GC. The FA composition was observed within the range of 25%-60.87% for saturated, 22.57%-56.45% for monounsaturated, and 5.64%-18.51% for total polyunsaturated FAs. The essential polyunsaturated fatty acid were higher in Stages I, IV, and V, suggesting a major role in embryogenesis. The cellular proliferation and organogenesis as visualized in the real-time imaging correlate well with the biochemical variations observed in C. pseudogracilirostris.

Molecular interaction of human acetylcholinesterase with trans-tephrostachin and derivatives for Alzheimer's disease.

Alzheimer's disease (AD), a neurodegenerative disorder affects more than 35 million people globally. Acetylcholinesterase suppression is the common approach to enhance the well-being of AD patients by increasing the duration of acetylcholine in the cholinergic synapses. Generally, herbal secondary metabolites are reported to be a major resource for acetylcholinesterase inhibitors (AChEIs). Trans-tephrostachin was reported from Tephrosia purpurea for AChE inhibition. Here, we report on the design, synthesis, and assessment of human acetylcholinesterase inhibitory activity from trans-tephrostachin derivatives or analogs as anti-AD agents. The five newly synthesized compounds 4a. 4b, 4c, 4d and 4e displayed potent inhibitory activities with IC50 values of 35.0, 35.6, 10.6, 10.3, and 28.1 µM respectively. AChE enzyme kinetic study was performed for the five derived compounds using the Ellman's method. The Lineweaver-Burk and the secondary plots revealed the mixed inhibition for 4a, 4c and 4d whereas 4b and 4e demonstrated competitive inhibition. Molecular docking and molecular dynamics simulations showed the derivatives or analogs of trans-tephrostachin attained a high binding affinity and efficacy than the standard drug. In conclusion, trans-tephrostachin and its derivative compounds could become effective agents for further drug development to treat AD.

Zebrafish as an Emerging Model for Bioassay-Guided Natural Product Drug Discovery for Neurological Disorders.

Most neurodegenerative diseases are currently incurable, with large social and economic impacts. Recently, there has been renewed interest in investigating natural products in the modern drug discovery paradigm as novel, bioactive small molecules. Moreover, the discovery of potential therapies for neurological disorders is challenging and involves developing optimized animal models for drug screening. In contemporary biomedicine, the growing need to develop experimental models to obtain a detailed understanding of malady conditions and to portray pioneering treatments has resulted in the application of zebrafish to close the gap between in vitro and in vivo assays. Zebrafish in pharmacogenetics and neuropharmacology are rapidly becoming a widely used organism. Brain function, dysfunction, genetic, and pharmacological modulation considerations are enhanced by both larval and adult zebrafish. Bioassay-guided identification of natural products using zebrafish presents as an attractive strategy for generating new lead compounds. Here, we see evidence that the zebrafish's central nervous system is suitable for modeling human neurological disease and we review and evaluate natural product research using zebrafish as a vertebrate model platform to systematically identify bioactive natural products. Finally, we review recently developed zebrafish models of neurological disorders that have the potential to be applied in this field of research.

Zebrafish bio-assay guided isolation of human acetylcholinesterase inhibitory trans-tephrostachin from Tephrosia purpurea (L.) Pers.

An acetylcholinesterase inhibitory compound was isolated from Tephrosia purpurea (L.) Pers. by zebrafish brain based bioassay guided isolation and predicted as trans-tephrostachin.Enzyme kinetics studies (Line weaver-Burk plots and Michaelis Menten equation) favored the reversible / mixed type, with the inhibition constant (Ki) of 53.0 ± 7.4 µM in zebrafish brain (IC50 value of 39.0 ± 1.4 µM). However, the inhibition constant (Ki) was found to be 36.0 ± 0.4 µM with IC50 value of 20.0 ± 1.0 µM, whereas donepezil showed 3.2 ± 0.3 µM with the IC50 value of 0.12 ± 0.04 µM for human acetylcholinesterase. Further, the molecular docking, dynamics and simulation for trans-tephrostachin obtained better binding affinity and efficacy than commercial drugs donepezil and galanthamine. Hence, the isolated compound trans - tephrostachin from T. purpurea shall be further considered for the development of potential drug for the counteraction of Alzheimer's disease progression.

Partial characterization of a biosurfactant extracted from Pseudomonas sp. B0406 that enhances the solubility of pesticides.

Biodegradation of some organochlorine and organophosphate pesticides is difficult because of their low solubility in water and, therefore, their low bioavailability. To overcome the hydrophobicity problem and the limited pesticide availability, biosurfactants play a major role. In this study, we evaluated the effect of an extract from Pseudomonas sp. B0406 strain with surfactant properties, on the solubility of two pesticides: endosulfan (ED) and methyl parathion (MP). Such a process was performed in order to increase the aqueous solubility of both pesticides, to increase its availability to microorganisms and to promote their biodegradation. The extract from Pseudomonas sp. B0406 showed a critical micellar concentration of 1.4 g/L and the surface tension at that point was 40.4 mN/m. The preliminary chemical and physical partial characterization of the extract with surfactant properties indicated that it is an anionic glycolipid, which increases the solubility of both pesticides of 0.41 at 0.92 mg/L for ED and of 34.58 at 48.10 mg/L for MP. The results of this study suggest the effectiveness of this extract in improving the solubility of both pesticides ED and MP in water and, therefore, of its potential use to enhance the degradation of these pesticides.

Molecular Dynamics Validation of Crizotinib Resistance to ALK Mutations (L1196M and G1269A) and Identification of Specific Inhibitors.

Anaplastic lymphoma kinase (ALK) positive non-small cell lung cancer (NSCLC) patients are mostly treated with ALK tyrosine kinase inhibitors (TKIs). Crizotinib is the first generation ALK inhibitor practiced as a primary chemo to combat cancer cells followed by second generation inhibitor ceritinib which are effective against crizotinib resistant ALK mutations. However, patients treated with these drugs invariably relapsed because of the development of new drug resistance mutations. In this study we explored the crizotinib resistance in the presence of ALK mutations L1196M and G1269A through molecular dynamics simulation studies. Further mutation specific inhibitors CID 71748211 and CID 71728095 were identified to potentially inhibit ALK with mutations L1196M and G1269A, respectively. This computational investigation in-sighted the molecular factors involved in crizotinib resistance which enhanced in the identification of new ALK drugs that brings individualized medicine to treat ALK positive NSCLC patients with specific mutations. J. Cell. Biochem. 118: 3462-3471, 2017. © 2017 Wiley Periodicals, Inc.

Rapid neurobehavioural analysis based on the effects of an acetylcholinesterase inhibitor from Tephrosia purpurea in Zebrafish.

BACKGROUND: Zebrafish is an important model organism for human neurobehavioural studies and compound screening for neurodegenerative disorders like Alzheimer's disease and dementia. PURPOSE: We wanted to analyse the rapid neurobehavioural effects based on acetylcholinesterase inhibitors from Tephrosia purpurea in zebrafish. METHODS: Tephrosia purpurea (a herbal neuroactive molecule) extract was analyzed for its rapid neurobehavioural effects and the corresponding psychotic twitches were quantified. The inhibitory activity of acetylcholinesterase was analysed in Zebrafish. The activity of the molecule was confirmed after column chromatography and RP-HPLC purifications. The toxicity of the molecule was also studied in developing zebrafish embryos. RESULTS: The psychotic body twitches were calculated as crude 484.67 ± 18.01 at 10 µg/mL, column purified 616 ± 9.64 at 8 µg/mL, Sep-Pak C18 712.67 ± 19.5 at 6 µg/mL, and HPLC elution showed 815 ± 14.93. The minimum inhibitory concentration of acetylcholinesterase in crude and column purified was 100 ng/mL and 5 ng/mL. The IC50 value of acetylcholinesterase inhibitor was calculated as 626ng/mL for crude (P<0.0001), 28.92 ng/mL for Sep-Pak column elution and 64 ng/mL for Donepezil. The HPLC fifth elution showed inhibition percentage as 96.97 ± 0.12. The organogenesis effects were seen in the drug concentration of 10 µg/mL. Pericardial bulging, trunk and tail flexure with heart edema and head edema were observed in the embryos at higher dose of 30- 40 µg. The LC50 value was 34.87 µg/mL. The studies showed that the inhibitory concentration of acetylcholinesterase is comparatively higher than Donepezil. CONCLUSION: Rapid behaviour based screening is an inexpesive assay to identify neuroactive small molecules. This herbal can be further used for the development of drugs for Alzheimer's disease.

Isolation of a small molecule with anti-MRSA activity from a mangrove symbiont Streptomyces sp. PVRK-1 and its biomedical studies in Zebrafish embryos

<p><b>OBJECTIVE</b>The aim of the present study was to isolate the anti-MRSA (Methicillin Resistant Staphylococcus aureus) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos.</p><p><b>METHODS</b>MRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC50 were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish.</p><p><b>RESULTS</b>The bioactive anti-MRSA small molecule A2 was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A2 was 30 µg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 µg/mL for TLC purified molecule A2 with LC50 mean value was 61.504 µg/mL. Zebrafish toxicity was assessed in 48-60 µg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 µg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A2 did not affected the HBR.</p><p><b>CONCLUSIONS</b>Anti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.</p>