Search

The experiments were carried out during February-July, 2021 at the Department of Livestock Products Technology (Meat science), Veterinary College and Research Institute, Namakkal, Tamil Nadu Veterinary and Animal Sciences University, India to assess the influence of heat on protein degradation, histology and eating quality indicators of chicken. Meat samples were taken from the broiler breast and cooked for the targeted end point temperature (75°C, 83°C, 89°C and 94°C) then assessed pH, colour, cooking loss, shear force value, protein degradation (SDS PAGE) and structure of muscle fibers. The heating temperature had a significant effect on pH and shear force value (p<0.01) at 94°C. Colour L* decreased significantly (p<0.01) and a* and b* value increased significantly (p<0.01) of cooked chicken breast at increasing end point cooking temperature. Progressive increasing (p<0.01) trends of cooking loss were noticed during the rising of end point cooking temperature. There were no significant differences between muscle fiber diameters. But significant (p<0.01) differences were observed in the sarcomere length of cooked chicken meat, showing decreasing trends during increasing temperature. Analysis of SDSPAGE showed that the intensity of the myofibrillar protein band gradually decreased during chicken meat cooking at a higher endpoint temperature and inferring that the meat protein had changed. The heating temperature had a significant effect on sensory quality of cooked chicken breast at increasing end point cooking temperature. This study revealed that meat cooking at high-end point temperatures affects the meat quality by degradation of muscle proteins and changes in muscle structure. [ABSTRACT FROM AUTHOR]

The utilization of underpriced fish pangas (Pangasianodon hypophthalmus) through salt fermentation was investigated. The fish was salt fermented, and its biochemical, sensory, and other quality characteristics determined for 180 days. Moisture and salt contents of the product toward the end of fermentation maintained at 54.34 and 16.54%, respectively, while the pH had dropped to 5.34. The first phase of fermentation continued up to 120 days where rapid changes in salt, pH, protein, and lipid degradation products were seen. This was followed by a stabilization phase till 150th day with minimal changes followed by the maturation stage where flavor components improved and many parameters were more stable than in the stabilization phase at the end. Slow increase in fat and protein degradation products like thiobarbituric acid (TBA), total volatile basic nitrogen (TVBN), free fatty acid (FFA), and peroxide value (PV) indicated controlled degradation of the macromolecules. At the end of 6 months of salt fermentation, product stability was found to be a result of hurdle effects of unfavorable intrinsic parameters. Practical applications: Application of salt curing on pangas will give new dimension to value addition of the fish and can make ways for better utilization and value realization of the underpriced fish. This could raise interest among the pangas growers for more production. The salt fermentation of pangas masks the original fatty flavor of the raw fish after fermentation and results in fermented flavor. This salt fermentation process could surely enhance the saleability value of pangas which can serve as an alternative to highly priced salt‐fermented hilsa in India and Bangladesh. [ABSTRACT FROM AUTHOR]

Ozonide antimalarials, OZ277 (arterolane) and OZ439 (artefenomel), are synthetic peroxide-based antimalarials with potent activity against the deadliest malaria parasite, Plasmodium falciparum. Here we used a "multi-omics" workflow, in combination with activity-based protein profiling (ABPP), to demonstrate that peroxide antimalarials initially target the haemoglobin (Hb) digestion pathway to kill malaria parasites. Time-dependent metabolomic profiling of ozonide-treated P. falciparum infected red blood cells revealed a rapid depletion of short Hb-derived peptides followed by subsequent alterations in lipid and nucleotide metabolism, while untargeted peptidomics showed accumulation of longer Hb-derived peptides. Quantitative proteomics and ABPP assays demonstrated that Hb-digesting proteases were increased in abundance and activity following treatment, respectively. Ozonide-induced depletion of short Hb-derived peptides was less extensive in a drug-treated K13-mutant artemisinin resistant parasite line (Cam3.IIR539T) than in the drug-treated isogenic sensitive strain (Cam3.IIrev), further confirming the association between ozonide activity and Hb catabolism. To demonstrate that compromised Hb catabolism may be a primary mechanism involved in ozonide antimalarial activity, we showed that parasites forced to rely solely on Hb digestion for amino acids became hypersensitive to short ozonide exposures. Quantitative proteomics analysis also revealed parasite proteins involved in translation and the ubiquitin-proteasome system were enriched following drug treatment, suggestive of the parasite engaging a stress response to mitigate ozonide-induced damage. Taken together, these data point to a mechanism of action involving initial impairment of Hb catabolism, and indicate that the parasite regulates protein turnover to manage ozonide-induced damage. Author summary: The ozonides are a novel class of fully synthetic antimalarial drugs with potent activity against all parasite species that cause malaria, including the deadliest, Plasmodium falciparum. With the emergence of resistance to current frontline artemisinin-based antimalarials, new drugs are urgently needed and a clear understanding of their mechanism of action is essential so that they can be optimally deployed in the field. Here, we studied the biochemical effects of two ozonides, OZ277 (marketed in India in combination with piperaquine) and OZ439 (in Phase IIb clinical trials) in P. falciparum parasites using an untargeted multi-omics approach consisting of proteomics, peptidomics and time-dependent metabolomics, along with activity-based protease profiling. We found that the ozonides initially disrupt haemoglobin metabolism and that they likely engage the parasite proteostatic stress response. Furthermore, when the duration of ozonide exposure was extended beyond 3 hours to reflect clinically-relevant exposure periods, additional parasite biochemical pathways were perturbed. This comprehensive analysis provides new insight into the antimalarial mode of action of ozonides and provides new opportunities for interventions to enhance their antimalarial efficacy. [ABSTRACT FROM AUTHOR]

Twenty four samples of kulfi, eight from each of the following sources viz. road side, small shops and organised sectors, were collected aseptically and subjected to estimation of total count, coliform count and psychrotrophic count to determine the microbiological quality of kulfi sold in Chennai. The level of psychrotrophic count was found to be more in road side local vendor samples (1.8 x 10³ cfu/g) followed by that of samples of small scale producers (1.01 x 103 cfu / g) and branded samples from organised sector (3.5 x 10² cfu / g). Road side local vendor samples contributed 56.90 % of organisms to the psychrotrophic count, followed by small scale producers (32.03 %), while organised sector contributed the least (11.07 %). Pseudomonas fluorescens was found to have greater proteolytic and lipolytic effect when compared with the activity of Pseudomonas aeruginosa and Pseudomonas putrifaciens. The mean total count and coliform count were found to be maximum in road side local vendor samples followed by samples of small scale producers The microbial quality of most of the kulfi samples needs to be improved in order to avoid any food poisoning and to safeguard the health of the consumers. [ABSTRACT FROM AUTHOR]

The study aims to assess the in vitro anti-inflammatory property of aqueous extracts of novel herbal formulations consisting of Phyllanthus niruri, Boerhavia diffusa and Picrorhiza kurroa (Liver Kidney Care™– LKC™,Organic India™) and Zingiber officinale (Ginger, Organic India™). The extracts were analyzed by Liquid Chromatography-Mass Spectrometry (LC–MS) for the detection of active ingredients. LC–MS confirmed the presence of active ingredients Phyllanthin, Boeravinone-B, Picroside I and Picroside II in LKC™; and 6-Gingerol in Ginger. Cytotoxicity assessments were done by Trypan blue cell exclusion method. The cells were pre-incubated with extracts from two herbal formulations LKC™or Ginger prior to TNFα or arachidonic acid (AA) stimulation to determine their effects on inflammatory signalling. Non-toxic doses of the extracts were used to determine their anti-inflammatory property by quantifying mRNA expression of IL-6, IKBA, ALOX5 and COX2 using qPCR and protein expression of IκBα and phospho-p65 by western blot. TNFα-induced mRNA expression of IL-6, IKBA and COX2 , and IκBα protein degradation along with phosphorylation of p65 was inhibited by LKC™and Ginger extracts. AA-induced mRNA expression of COX2 , ALOX5 and IL-6 , and phosphorylation of p65 was inhibited by LKC™and Ginger extracts.LKC™ and Ginger aqueous extracts inhibited TNFα- or AA-induced intracellular inflammatory signalling pathways (NFκB, cyclooxygenase, lipooxygenase),suggesting the anti-inflammatory potential of these herbal formulations. [ABSTRACT FROM AUTHOR]

A young Indian female visited hospital as a suspected case of thrombotic thrombocytopenic purpura (TTP) with relapsed thrombotic complications with low platelet counts, infarct in middle cerebral artery and thrombi in microvessels. We first confirmed the deficiency of ADAMTS13 metalloprotease in this patient showing improper cleavage of vWF multimers by her plasma unlike her parents and brother. Although patient had very less ADAMTS13 antigen in plasma, but it did not appear to be the cause of deficiency of the enzyme, because her father had similarly low antigen level and he never had prothrombotic complications. While investigating the genetic change in ADAMTS13, we observed four homozygous-SNPs (g.420T>C, g.1342C>G, g.1716G>A and g.2280T>C) in exon 5, 12, 15 and 19 respectively in patient and her father unlike the heterozygous form of same SNPs in mother and brother. Further to investigate the cause of ADAMTS13 deficiency, we observed an elevated level of antibody against ADAMTS13 in patient unlike her father and other family members. Our study therefore provides the molecular approach of diagnosis of TTP in this patient and also highlights the use of such techniques in India. More importantly, study provides the clue of alternate treatment such as immunosuppressant therapy to this patient., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Human immunodeficiency virus 1 (HIV-1) diversity is a significant challenge in developing a vaccine against the virus. B/C recombinants have been found in India and other places but are the predominant clade prevalent in China. HIV-1 envelopes (Envs) are the target of broadly neutralizing antibodies (bNAbs) which develop spontaneously in some HIV-1 infected patients. It has been previously reported with efficiently cleaved clade A, B and C Envs that preferential binding of Envs to bNAbs as opposed to non-NAbs, a desirable property for immunogens, is correlated with efficient cleavage of the Env precursor polypeptide into constituent subunits. These Envs are suitable for designing immunogens as soluble proteins, virus-like particles or for delivery by viral vectors/plasmid DNA. However, a B/C recombinant Env with similar properties has not been reported. Here we show that the chimeric, recombinant B/C clade Env LT5.J4b12C is efficiently cleaved on the plasma membrane and selectively binds to bNAbs.

Protease enzymes in tannery industries have enormous applications. Seeking a potential candidate for efficient protease production has emerged in recent years. In our study, we sought to isolate proteolytic bacteria from tannery waste dumping site in Tamilnadu, India. Novel proteolytic Bacillus alkalitelluris TWI3 was isolated and tested for protease production. Maximum protease production was achieved using lactose and skim milk as a carbon and nitrogen source, respectively, and optimum growth temperature was found to be 40 °C at pH 8. Protease enzyme was purified using ammonium sulfate precipitation method and anion exchange chromatography. Diethylaminoethanol (DEAE) column chromatography and Sephadex G-100 chromatography yielded an overall 4.92-fold and 7.19-fold purification, respectively. The 42.6-kDa TWI3 protease was characterized as alkaline metallo-protease and stable up to 60 °C and pH 10. Ca(2+), Mn(2+), and Mg(2+) ions activated the protease, while Hg(2+), Cu(2+), Zn(2+), and Fe(2+) greatly inhibited it. Ethylenediaminetetraacetic acid (EDTA) inhibited TWI3 protease and was activated by Ca(2+), which confirmed that TWI3 protease is a metallo-protease. Moreover, this protease is capable of dehairing goat skin and also removed several cloth stains, which makes it more suitable for various biotechnological applications.

Loss of function of TAR DNA-binding protein (TDP-43) has been implicated in neurodegenerative disorders in both humans and animal models. TDP-43 has also been shown to be cis-acting transcriptional repressor of the acrosome vesicle (Acrv) gene in mice. In the present study, we investigated the expression of the TDP-43 transcript (TARDBP) and protein in germ cells from 11 fertile and 98 subfertile men to verify its potential association with poor seminograms. The expression profile of TDP-43 was characterised in immature germ cells and spermatozoa from semen from fertile and subfertile men using reverse transcription-polymerase chain reaction, western blotting and immunofluorescence. Although germ cells from subfertile men tested negative for TARDBP, the full-length message of the same was detected in fertile men. TDP-43 was detected in spermatozoa from fertile men using western blot analysis and immunofluorescence. The expression of this protein was negligible in spermatozoa from men with primary spermatogenic dysfunction. We conclude that a deficiency in the TDP-43 expression is associated with defective spermatogenesis and male infertility. We propose that TDP-43 could be used as a marker of male factor infertility.

Mass spectrometry-based amino acid sequencing is currently based almost entirely on collision-induced peptide fragmentation and analyses. Here, we describe a single-stage MS-based technique for amino acid sequencing involving partial, heterogenous digestion of a peptide by a processive, non-specific, heat-loving Bacillus subtilis-derived aminopeptidase (BsuAP), which acts optimally at 70 °C and allows 'single-shot' sequencing to be carried out through simultaneous accumulation, and detection of sub-populations of peptides of progressively reducing length., (Copyright © 2015 Elsevier Inc. All rights reserved.)

A nutrition survey carried out in India revealed that the diets of the rural population are inadequate and deficient in most of the nutrients especially protein. India being the 5th-largest producer of soybean, a protein-rich cereal, can redress protein-energy malnutrition through diversification of soybean uses by developing high-value and health-based food products. Tofu, a nonfermented soybean product rich in high-quality protein, B-vitamins, and isoflavones, could be an excellent substitute for meat in Indian recipes. Tofu being rich in protein has a very short shelf life. Hence an attempt was made to improve the shelf life using extracts of tulsi ( Ocimum sanctum) commonly available in rural areas. Tofu was prepared traditionally using MgCl2:CaSO4 as coagulating agents. Aqueous extract of Ocimum sanctum (tulsi) was added during the preparation and storage of tofu to prolong its shelf life. Water used in this study was free from microflora, plant extract used contained mesophilic count of 2.527 × 104 CFU/g, and no yeasts and molds were detected. Tofu with tulsi extract had 76.4% moisture and was softer than control. Not much difference in mesophilic count was observed between control and treated samples during storage; however, treated tofu was organoleptically good until the end of the study with less lipid-peroxidation and exhibited 50% (4.7 units) less protease activity than control (9.6 units) after 7 d. By using extracts of naturally available, easily cultivable tulsi, the shelf life was successfully extended to 7 to 8 d from 3 to 4 d of normal storage without refrigeration. [ABSTRACT FROM AUTHOR]

Background: In the present investigation, the effect of Aloe vera gel powder on angiotensin-converting enzyme (ACE) inhibitory activity, extent of proteolysis during fermentation and survival of Lactobacillus casei NCDC19 during storage of fermented milk was studied., Results: Among the different cultures screened for ACE inhibitory activity, Lactobacillus casei NCDC 19 exhibited the highest ACE inhibition (approx. 40%) as well as extent of proteolysis (0.37, Abs₃₄₀). In the presence of Aloe vera (0.5% and 1% w/v) an increase in extent of proteolysis (0.460 ± 0.047 and 0.480 ± 0.027) and percent ACE inhibitory activity (44.32 ± 2.83 and 47.52 ± 1.83) was observed in comparison to control. Aloe vera powder addition also led to an increase in viable counts (>11 log cfu mL⁻¹) of L. casei NCDC 19 in fermented milk during storage for 7 days and the counts were maintained in sufficiently higher numbers., Conclusion: The study suggests Aloe vera to be a good functional ingredient which can be further explored for different health attributes., (© 2014 Society of Chemical Industry.)

Matrix metalloproteinases (MMPs) are the major endopeptidases involved in proteolysis of blood brain barrier (BBB) during central nervous system (CNS) infections. The present study detected serum levels and activities of MMP-2 and MMP-9 in patients with neurocysticercosis (NCC) and their association with symptomatic disease. In total, 68 individuals with NCC (36 symptomatic patients with active seizures and 32 asymptomatic individuals) and 37 healthy controls were enrolled for the study. Serum MMP-2 and MMP-9 levels and their activities were measured by ELISA and gel zymography respectively. Mean serum MMP-2 levels (ng/ml) were higher both in asymptomatic and symptomatic NCC cases compared to healthy controls. However, significantly higher levels of serum MMP-9 (ng/ml) were detected only in symptomatic NCC patients compared to asymptomatic NCC cases and healthy controls. Levels of both MMPs positively correlated with symptomatic NCC. Serum MMP-2 activities were significantly higher in symptomatic and asymptomatic NCC compared to healthy controls whereas serum MMP-9 activity was significantly associated with symptomatic NCC compared to healthy controls and asymptomatic NCC. In conclusion, the elevated level of MMP-9 in serum appears to play an important role in the development of symptoms i.e. active seizures in patients with NCC. However, further studies are needed to elucidate its precise role in disease pathogenesis.