Immunobiochemical Characteristics of Purified Native Leptin Protein from Indian Major Carp, Rohu (<i>Labeo rohita</i>Ham.)

Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita), at molecular level, the present study was designed to isolate rohu leptin from its hepatocytes (the prime source of leptin in fish) and immunobiochemical characterization of the same, subsequently. In the present study, chemical treatment and ultra-sonication technique was used for isolating leptin from rohu liver tissue. Purification of the protein was attempted using affinity column chromatography. The molecular, biophysical and serological characterization of rohu leptin was carried out by 2D-gel electrophoreis, SDS-PAGE, MALDI-TOF Mass spectroscopy and Western blot. The SDS-PAGE and 2D gel analysis revealed that rohu native leptin possesses molecular mass of 16 kDa. Western blot analysis showed that the fish hepatocytes possessed the sero-reactive leptin protein of 16 kDa. MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of rohu leptin as 16283.38 Da. The serodiagnostic potential of native leptin of rohu was revealed for the first time while assessing its serological responses by ELISA using anti-leptin antibodies.

Low doses of ethanolic extract of Boldo(Peumus boldus) can ameliorate toxicity generated by cisplatin in normal liver cells of mice in vivo and in WRL-68 cells in vitro, but not in cancer cells in vivo or in vitro

OBJECTIVE： Use of cisplatin, a conventional anticancer drug, is restricted because it generates strong hepatotoxicity by accumulating in liver. Therefore its anticancer potential can only be fully exploited if its own toxicity is considerably reduced. Towards this goal, ethanolic extract of the plant, Boldo （Peumus boldus）, known for its antihepatotoxic effects, was used simultaneously with cisplatin, to test its ability to reduce cisplatin＇s cytotoxicity without affecting its anticancer potential. METHODS： The cytotoxicity of Boldo extract （BE） and cisplatin, administered alone and in combination, was determined in three cancer cell lines （A549, HeLa, and HepG2） and in normal liver cells （WRL-68）. Drug-DNA interaction, DNA damage, cell cycle, apoptosis, reactive oxygen species （ROS） and mitochondrial membrane potential （MMP, Aψ） were also studied. Hepatotoxicity and antioxidant activity levels were determined by alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and glutathione assays in mice. The cytotoxicity of related proteins was tested by Western blotting. RESULTS： Co-administration of BE and cisplatin increased viability of normal cells, but had no effect on the viability of cancer cells. Boldo protected liver from damage and normalized different antioxidant enzyme levels in vivo and also reduced ROS and re-polarized MMP in vitro. Bax and cytochrome c translocation was reduced with caspase 3 down-regulation. Further, a drug- DNA interaction study revealed that BE reduced cisplatin＇s DNA-binding capacity, resulting in a reduction in DNA damage. CONCLUSION： Results indicated that a low dose of BE could be used beneficially in combination with cisplatin to reduce its toxicity without hampering cisplatin＇s anticancer effect. These findings siqnify a potential future use of BE in cancer therapy.

First report on chlorophyllin to protect mammalian and fish muscle cells from pesticide toxicity via activation of p53 and PARP

Objectives:Pesticide toxicity has become one of the major environmental menaces affecting all types of life forms of the ecosystem.Pesticides get washed off from agricultural fields into nearby water bodies and enter the aquatic organisms.Their bio-accumulated form finally reaches the human race,through consumption of pesticide infested aquatic animals,causing several physiological dysfunctions.Hence it becomes necessary to find a therapeutic cure/a preventive measure to stop the health hazard issues of pesticide.With this projection a search for a phyto-based-product was made whose primary objective would be to lower the pesticidal toxicity in fish and simultaneously in the human race.Methods:In this study we tried to check whether the phyto-chemical,Chlorophyllin(CHL),known for its anti-genotoxic,anti-oxidant activities,could render any kind of protection against Cypermethrin(CM)induced-toxicity in fish model and mammalian cell line L6.Both the model L6 and fish were pre-treated with CHL prior to exposure of CM.Different scientific parameters like%cellular cytotoxicity,reactive oxygen species(ROS)generation,nuclear condensation,etc were checked to validate the possibility of CHL in protecting CM-induced toxicity.Results:The overall results revealed that pre-treatment with CHL could restrict the ROS generation leading to modulation in associated cytokine proteins expression NFkβand IFNγ.Further,CHL lowered nuclear condensation and elevated expression of DNA repair proteins p53 and PARP,showing a kind of pre-activation of signalling cascades for overall protection against the severity of pesticidal toxicity.Conclusion:Thus,this phyto-based preventive approach would possibly solve many areas of human health issues related to pesticide toxicity in future.