Methanolic extract of Phlogacanthus thyrsiflorus Nees leaf induces apoptosis in cancer cells

Phlogacanthus thyrsiflorus Nees is a medicinal herb commonly used in traditional folk medicine, and it is known to possess anticancer activity. Here, we explored the anticancer properties of methanolic extract of P. thyrsiflorus leaves in HeLa and MCF-7 cell lines. We observed nuclear fragmentation as indication of apoptosis in the MPT treated cancer cells using haematoxylin and eosin (H&E) as well as fluorescent dye. DNA from the treated cells showed characteristic laddering of DNA fragments in agarose gel electrophoresis. Cell populations undergoing secondary necrosis following apoptosis could also be detected in FACS by annexin V/propidium iodide (PI) staining. Activated caspase-3 in the treated HeLa cells was detected by polyclonal anti-caspase-3 antibody utilizing immunocytochemistry. Using transmission electron microscopy, sub-cellular changes like rough endoplasmic reticulum, swollen mitochondria, distorted mitochondrial membrane, loss of cristae and matrix were observed in the treated HeLa cells. Extensive plasma membrane blebbing was also observed by scanning electron microscopy. Our findings support that Phlogacanthus thyrsiflorus leaves are natural source of potent anticancer agent

Methanolic extract of Phlogacanthus thyrsiflorus Nees leaf induces apoptosis in cancer cells

153-161Phlogacanthus thyrsiflorus Nees is a medicinal herb commonly used in traditional folk medicine, and it is known to possess anticancer activity. Here, we explored the anticancer properties of methanolic extract of P. thyrsiflorus leaves in HeLa and MCF-7 cell lines. We observed nuclear fragmentation as indication of apoptosis in the MPT treated cancer cells using haematoxylin and eosin (H&E) as well as fluorescent dye. DNA from the treated cells showed characteristic laddering of DNA fragments in agarose gel electrophoresis. Cell populations undergoing secondary necrosis following apoptosis could also be detected in FACS by annexin V/propidium iodide (PI) staining. Activated caspase-3 in the treated HeLa cells was detected by polyclonal anti-caspase-3 antibody utilizing immunocytochemistry. Using transmission electron microscopy, sub-cellular changes like rough endoplasmic reticulum, swollen mitochondria, distorted mitochondrial membrane, loss of cristae and matrix were observed in the treated HeLa cells. Extensive plasma membrane blebbing was also observed by scanning electron microscopy. Our findings support that Phlogacanthus thyrsiflorus leaves are natural source of potent anticancer agent

Antimicrobial peptides of buffalo and their role in host defenses

Antimicrobial peptides (AMPs) are highly conserved components of the innate immune system found among all classes of life. Buffalo (Bubalus bubalis), an important livestock for milk and meat production, is known to have a better resistance to many diseases as compared to cattle. They are found to express many AMPs such as defensins, cathelicidins, and hepcidin which play an important role in neutralizing the invading pathogens. Buffalo AMPs exhibit broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria. Similar to its natural form, synthetic analogs of buffalo AMPs are also antimicrobial against bacteria and even fungus making them a good target for the development of therapeutic antimicrobials. In addition to its antimicrobial effect, AMPs have been demonstrated to have a number of immunomodulatory functions, and their genes are responsive to infections. Further, induction of their gene expression by external factors may help in preventing infectious diseases. This review briefly discusses the AMPs of buffalo identified to date and their possible role in innate immunity

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Not AvailableH5N1 causes high mortality in domestic poultry. It is the causative agent of H5N1 flu and is the world's largest current pandemic threat. Haemagglutinin (HA) is a surface glycoprotein of the virus which facilitates viral attachment to target cell and its entry. HA frequently accumulates mutation to escape immune response of host. In this study, conserved amino acid sequences in HA2 subunit of H5N1 HA protein of both clade 2.2 and 2.3 were identified and the conserved sequence was further analyzed in silico for its antigenic index, hydrophilicity and surface probability. Six peptides showing potential antigenicity were selected and synthesized. Reactivity of the peptides were analysed by indirect ELISA using antisera against different subtypes of avian influenza. Five out of the six peptides showed reactivity. Thus, five epitopes in the conserved region of HA2 subunit of H5 could be identified which can detect positive serum against avian influenza.Not Availabl

Molecular Cloning and Expression Profile of Toll-Like Receptor 3 from an Indian Coldwater Fish, <i>Schizothorax richardsonii</i>(Gray)

<p>The innate immune system recognizes pathogens through pattern recognition receptors (PRRs), and toll-like receptors (TLRs) are one of the most important PRRs. TLR3 is a unique member of TLR family that recognizes double-stranded RNA (dsRNA), a viral replication intermediate. There is a variation in its response among diverse fish species toward the same stimulants. We identified and cloned TLR3 from Indian snow trout, Schizothorax richardsonii and carried out its expression analysis in un-induced and poly (I:C) challenged fish. It has an open reading frame (ORF) of 2712 bases that encodes a polypeptide of 904 amino acids. The molecular weight of the polypeptide was predicted to be 102.4482 kDa with an isoelectric point of 7.40. Quantitative real time PCR (qRT-PCR) was carried out after 24 hours of poly (I:C) treatment and expression of TLR3 was analyzed in different tissues. As compared with untreated fish the poly (I:C) challenged fish revealed significantly high expression of TLR3 in kidney followed by liver and gills.</p

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Not AvailableKisspeptin 1 is a neuropeptide hormone of the RFamide family, which act as an upstream regulator of brain-pituitary-gonad (BPG) axis in most vertebrates including teleosts. In the present study, a 16 amino acid long putative mature bioactive peptide (kiss 1) from preprokisspeptin 1 of golden mahseer, Tor putitora (Hamilton, 1822), was synthesized and characterized using an integrated (experimental and in silico) approach. The far-UV circular dichroism (CD) spectrum of this peptide was evaluated both in aqueous and membrane mimicking solvents (TFE, HFIP and Dioxane). The results indicate that kiss 1 peptide adopted helical, turn and β conformations in membrane like environments. The near-UV CD spectroscopy was also carried out to examine the tertiary packing around aromatic residues of kiss 1 peptide and the peptide-membrane complex. The kiss 1 peptide exhibited little signal in water, but a prominent negative band was observed at around 275 nm when membrane mimetic solution was added. The observed ordered conformations of kiss 1 peptide in the different solvents indicated its potential biological activity which could enhance the secretion of gonadotropin-releasing hormone (GnRH) at BPG axis. The conformational information generated from the present study reinforces the application prospects of bioactive synthetic peptide analogs of kisspeptin 1 in improving the reproductive performances of important cultivable fish species.Indian Council of Agricultural Research (ICAR

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Not AvailableRabies virus (RABV) is neurotropic and infects all warm-blooded animals. The binding of the virus with host cell receptor components is critical for infection. The present study reports the interaction of nicotinic acetylcholine receptor alpha 1 (nAChRα1) peptides with the rabies virus glycoprotein (RABVG) to design potential anti-rabies agents. The nAChRα1peptide sequences from different species (bovine, human and electric fish/torpedo) were synthesized and their secondary structures were characterized using CD spectroscopy. The molecular docking analysis of nAChRα1 peptides with RABVG indicated the involvement of specific domains and their particular amino acid contributions. Bovine peptide (C-32) (docking score of 14146 kJ/mol) and torpedo peptide (T-32) (docking score of 13704 kJ/mol) were found to interact strongly with RABVG. T-32 peptides had the highest binding and inhibiting property against RABV compared to other peptide sequences. The results of both computational and experimental methods demonstrated that nAChRα1 peptides and their analogs may serve as potential antiviral agents against RABV infection.Not Availabl

Conformational analysis of a synthetic fish kisspeptin 1 peptide in membrane mimicking environments - Fig 3

<p>Secondary structure analysis of kiss 1 peptide using online PSI-PRED (A) and GOR4 (B) tools.</p

Far-UV CD spectra of kiss 1 peptide in water and TFE.

<p>(a) CD spectrum in water. (b) CD spectrum in 45% TFE. (c) CD spectrum in 90% TFE.</p