Evaluation of Neutralization Potential of Naja naja and Daboiarusselii Snake Venom by Root Extract of Cyanthillium cinereum.

Aim: One of the main reasons for the death due to snake bites is the non-availability of antivenoms in the regions where they are needed. The use of medicinal plants and plant-based natural products as an alternative to antivenom will become a milestone in snake bite envenomation. The present study investigates the in vitro antivenom properties of Cyanthillium cinereum root extracts. Materials and methods: The C. cinereum root's aqueous extract was prepared by the Soxhlet extraction method, and phytochemical screening was performed to detect the presence of various bioactive compounds. Thin-layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) analysis were performed for the detection and identification of phytochemical constituents. In this study, an in vitro model is used to assess the antivenom capability of aqueous extract. Venom toxicity and neutralization assays were as follows: An in vitro pharmacological evaluation was performed by direct hemolysis assay, indirect hemolytic assay, proteolytic activity, neutralization of procoagulant activity, and gelatin liquefaction method. Results: Qualitative analysis of phytochemicals by the standard method showed the presence of various phytochemical constituents. Also, GC-MS analysis showed the presence of three major compounds that possess antivenom activity from the obtained 60 bioactive compounds, and their chemical structures were also determined. Venom protein profiling was performed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The plant extract was able to neutralize the Naja naja (N. naja) and Daboia russelii (D. russelii) venom induced hemolysis and it was reduced below 50 and 40%, respectively and the extract was also able to reduce the hemolytic halo produced by venoms. Procoagulant activity and gelatin liquefaction assay showed that venom-induced clotting was neutralized by increasing the root extract concentration sufficiently. Conclusion: The aqueous extract of the root of C. cinereum showed potent in vitro venom-neutralizing activity, and it can be used as a formidable therapeutic agent against N. naja and D. russelii envenomation. How to cite this article: Suji S, Dinesh MD, Keerthi KU, Anagha KP, Arya J, Anju KV. Evaluation of Neutralization Potential of Naja naja and Daboia russelii Snake Venom by Root Extract of Cyanthillium cinereum. Indian J Crit Care Med 2023;27(11):821-829.

Dopamine-induced photoluminescence quenching of bovine serum albumin-capped manganese-doped zinc sulphide quantum dots.

The direct detection of dopamine (DA) in human body fluids is a great challenge for medical diagnostics of neurological disorders like Parkinson's disease, Alzheimer's disease, senile dementia, and schizophrenia. In this work, a simple and turn off luminescence sensing of DA based on bovine serum albumin (BSA)-capped manganese-doped zinc sulphide quantum dots (Mn:ZnS/BSA QDs) is developed. The Mn:ZnS/BSA QDs were synthesized by a chemical co-precipitation method. Due to the special interaction of DA with BSA and metal ions, Mn:ZnS/BSA QDs can serve as an effective sensing platform for DA. The luminescence of Mn:ZnS/BSA QDs decreased linearly with increasing concentration of DA in the range from 6.6 to 50.6 nM. The limit of detection is 2.02 nM. The driving force for the luminescence quenching is partly provided by ground-state complex formation of QDs with DA. The photo-induced electron transfer from the conduction band of QDs to oxidized dopamine (quinone) also favors quenching. The Mn:ZnS/BSA QDs are barely interfered with by other competing biomolecules except catecholamine neurotransmitter like epinephrine. Moreover, this method is used in the analysis of DA-spiked human serum and human urine samples and good recovery percentages are found. To assess the utility of the developed sensor, paper strip assay was also successfully conducted. Graphical abstract.

Evaluation of DNA methylation and mRNA expression of heat shock proteins in thermal manipulated chicken.

Thermal manipulation during embryogenesis has been demonstrated to enhance the thermotolerance capacity of broilers through epigenetic modifications. Heat shock proteins (HSPs) are induced in response to stress for guarding cells against damage. The present study investigates the effect of thermal conditioning during embryogenesis and thermal challenge at 42 days of age on HSP gene and protein expression, DNA methylation and in vitro luciferase assay in brain tissue of Naked Neck (NN) and Punjab Broiler-2 (PB-2) chicken. On the 15th day of incubation, fertile eggs from two breeds, NN and PB-2, were randomly divided in to two groups: control (C)-eggs were incubated under standard incubation conditions, and thermal conditioning (TC)-eggs were exposed to higher incubation temperature (40.5°C) for 3 h on the 15th, 16th, and 17th days of incubation. The chicks obtained from each group were further subdivided and reared under different environmental conditions from the 15th to the 42nd day as normal [N; 25 ± 1 °C, 70% relative humidity (RH)] and heat exposed (HE; 35 ± 1 °C, 50% RH) resulting in four treatment groups (CN, CHE, TCN, and TCHE). The results revealed that HSP promoter activity was stronger in CHE, which had lesser methylation and higher gene expression. The activity of promoter region was lesser in TCHE birds that were thermally manipulated at the embryonic stage, thus reflecting their stress-free condition. This was confirmed by the lower level of mRNA expression of all the HSP genes. In conclusion, thermal conditioning during embryogenesis has a positive impact and improves chicken thermotolerance capacity in postnatal life.

T-homoeolog specific plasma membrane protein 3 [Nt(t)PMP3-2] in polyploid Nicotiana tabacum shows conserved alternative splicing, derived from extant Nicotiana tomentosiformis parent.

Abiotic stress induced plasma membrane protein 3 (PMP3) genes occur as multigene families in plants, coding for hydrophobic proteins. Group I PMP3s code for shorter ORFs while Group II PMP3s code for proteins with C-terminal extensions. Allotetraploid Nicotiana tabacum (SSTT; 2n = 48) derives its parentage from extant ancestors related to Nicotiana sylvestris (SS) and Nicotiana tomentosiformis (TT). Polyploidization triggers complex genetic and epigenetic changes, often leading to homoeolog-specific retention or loss of function, sub-functionalization or neo-functionalization. Genomic sequences of Nt(t)PMP3-1/Nt(t)PMP3-2 cloned from N. tabacum show near identity with N. tomentosiformis NtoPMP3-1/NtoPMP3-2 genomic sequences respectively (distinct from N. sylvestris NsPMP3-1/NsPMP3-2 genomic regions). RT-PCR with exon 1,2 primer pairs amplified only single fragments for Nt(t)PMP3-1 and Nt(t)PMP3-2. In contrast, for Nt(t)PMP3-2, three variants were detected using exon 2,3 primers by RT-PCR. Cloning revealed (i) a transcript coding for a Group I PMP3 [Nt(t)PMP3-2CS], (ii) a transcript with complete retention of the second intron [Nt(t)PMP3-2IR] and (iii) a transcript with an alternative (exon 2) 5' splice site [Nt(t)PMP3-2AS], coding for a longer protein, similar to ORFs of Group II PMP3 genes. All three Nt(t)PMP3-2 variants have conserved counterparts in the N. tomentosiformis transcriptome, suggesting the transcriptional machinery governing alternative splicing of Nt(t)PMP3-2 in N. tabacum has conserved origins, derived from a N. tomenosiformis lineage. The above data shows alternative splicing of PMP3 genes contributes to transcript and ORF diversity in plants. All three Nt(t)PMP3-2 splice variants show increased root-specific expression. Implications of Nt(t)PMP3-2 alternative splicing on transcript stability and ORF features are discussed.