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Rice cultivation generates huge amounts of on farm residues especially under mechanical harvesting. Paddy straw being recalcitrant hinders sowing of upcoming rabi crops like wheat and mustard. Non-environmental sustainable practice of on-farm burning of the paddy residues is being popularly followed for quick disposal of the agro-residues and land preparation. However, conservation agriculture involving in situ residue incorporation can be a sustainable option to utilize the residues for improvement of soil biological health. However, low temperature coupled with poor nitrogen status of soil reduces the decomposition rate of residues that may lead to nitrogen immobilization and hindrance in land preparation. In this direction, ecological impact of two approaches viz priming with urea and copiotrophic fungus-based bioformulation (CFB) consisting of Coprinopsis cinerea LA2 and Cyathus stercoreus ITCC3745 was studied for in situ degradation of residues. Succession of bacterial diversity was deciphered through high throughput whole metagenomic sequencing along with studies on dynamics of soil microbial enzymes. Treatments receiving CFB (T1) and urea (T2) when compared with bulk soil (absolute control) showed an increase in richness of the microbial diversity as compared to control straw retained treatment control (T3). The ß diversity indices also indicated sufficient group variations among the treatments receiving CFB and urea as compared to only straw retained treatment and bulk soil. Priming of paddy straw with CFB and urea also induced significant rewiring of the bacterial co-occurrence networks. Quantification of soil ligno-cellulolytic activity as well as abundance of carbohydrate active enzymes (CAZy) genes indicated high activities of hydrolytic enzymes in CFB primed straw retention treatment as compared to urea primed straw retention treatment. The genomic insights on effectiveness of copiotrophic fungus bioformulation for in situ degradation of paddy straw will further help in developing strategies for management of crop residues in eco-friendly manner.
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BACKGROUND: Prevalence of hepatitis C virus (HCV) infection among people who inject drugs in the state of Manipur, India, is 43%; however, access to care is poor. We piloted a Community-led and comprehensive hepatitis care model that included same-day HCV treatment at drug treatment centres. METHODS: Screening was conducted through venipuncture samples collected by community peer PWID, using HCV antibody (HCV Ab) rapid screening and hepatitis B virus (HBV) surface antigen (HBsAg) rapid diagnostic tests. Reactive HCV Ab samples were tested for HCV RNA using near point-of-care Truenat® HCV on Truelab® Quattro. Eligible HCV RNA-positive participants were treated on the same day using direct-acting antivirals and followed for sustained virologic response (SVR). HBsAg-negative participants received rapid HBV vaccination regimen while those positive for HBsAg were tested for DNA and referred for treatment. RESULTS: Between November 2021 and August 2022, 643 individuals were approached and 503 consented and were screened. All screened were males with history of injection drug use, and a median age of 27 years (IQR 23-32). Of the 241 (47.9%) HCV Ab reactive all underwent RNA testing and 156 (64.7%) were RNA detectable. Of those with viraemia, 155 (99.4%) were initiated on treatment with 153 (98.1%) on same day, with 2 (1.2%) HBsAg positive and waiting for HBV DNA results. Among those 153, median time from HCV Ab screening to treatment was 6 h 38 min (IQR 5 h 42 min-8 h 23 min). In total 155 (100%) completed HCV treatment, of those 148 (95.5%) completed SVR testing and 130 (87.8%) achieved SVR12. 27 (5%) participants were HBsAg-positive, 3 (11.1%) were also living with HCV viraemia; 443 (97.6%) were eligible for vaccination and 436 (98.4%) received all 3 vaccine doses. CONCLUSION: Community-led hepatitis care incorporating same day "test and treat" for HCV was feasible and effective. HBV screening identified a large proportion who were unvaccinated. Peer support extended resulted in ensuring compliance to care and treatment cascade and completing all the three doses of HBV vaccination. As the screening, diagnostics infrastructure and vaccine are available in most countries with national viral hepatitis programs also in place, our model can be adapted or replicated to progress towards global elimination targets.
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Estudos de Viabilidade , Grupo Associado , Abuso de Substâncias por Via Intravenosa , Humanos , Masculino , Abuso de Substâncias por Via Intravenosa/complicações , Adulto , Índia/epidemiologia , Adulto Jovem , Antivirais/uso terapêutico , Hepatite C/tratamento farmacológico , Anticorpos Anti-Hepatite C/sangue , Programas de Rastreamento/métodos , Antígenos de Superfície da Hepatite B/sangue , Projetos Piloto , Resposta Viral SustentadaRESUMO
KEY MESSAGE: We summarise recent advancements to achieve higher homologous recombination based gene targeting efficiency in different animals and plants. The genome editing has revolutionized the agriculture and human therapeutic sectors by its ability to create precise, stable and predictable mutations in the genome. It depends upon targeted double-strand breaks induction by the engineered endonucleases, which then gets repaired by highly conserved endogenous DNA repair mechanisms. The repairing could be done either through non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways. The HDR-based editing can be applied for precise gene targeting such as insertion of a new gene, gene replacement and altering of the regulatory sequence of a gene to control the existing protein expression. However, HDR-mediated editing is considered challenging because of lower efficiency in higher eukaryotes, thus, preventing its widespread application. This article reviews the recent progress of HDR-mediated editing and discusses novel strategies such as cell cycle synchronization, modulation of DNA damage repair factors, engineering of Cas protein favoring HDR and CRISPR-Cas reagents delivery methods to improve efficiency for generating knock-in events in both plants and animals. Further, multiplexing of described methods may be promising towards achieving higher donor template-assisted homologous recombination efficiency at the target locus.
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Sistemas CRISPR-Cas , Edição de Genes , Animais , Humanos , Edição de Genes/métodos , Recombinação Homóloga , Reparo do DNA/genética , Reparo de DNA por Recombinação , Reparo do DNA por Junção de ExtremidadesRESUMO
BACKGROUND AND AIMS: The Liver Frailty Index (LFI) is a well-studied tool that evaluates frailty in patients with cirrhosis. Consisting of grip strength, chair stands, and balance testing, the LFI has been associated with increased mortality in patients awaiting liver transplant. We aimed to extend our understanding of frailty in cirrhosis by exploring the relationship between the LFI and the risk of (1) cirrhosis progression, (2) mortality, and (3) unplanned hospitalizations, in both compensated and decompensated disease. APPROACH AND RESULTS: Adult patients with cirrhosis from four centers in North America and one in India were included. Frailty was measured at baseline using the LFI and categorized as robust (LFI < 3.2), prefrail (LFI 3.2-4.5), and frail (LFI > 4.5). Progression of cirrhosis was defined by an increase in clinical stage, ranging from 1 to 5, from baseline using the D'Amico classification. Factors associated with progression, mortality, and hospitalizations were evaluated using multivariate regression models, with transplant as a competing risk. In total, 822 patients with cirrhosis were included. Average Model for End-Stage Liver Disease (MELD) score was 15.5 ± 6.0. In patients with compensated cirrhosis, being frail versus robust was associated with increased risk of progression to the next cirrhosis stage or to death (HR, 2.45; 95% CI, 1.14-5.29) and with an increased risk of unplanned hospitalizations (2.32; 95% CI, 1.13-4.79), after adjusting for age, sex, and MELD score. Similar HRs were observed in patients with decompensated cirrhosis. CONCLUSIONS: Frailty was an independent predictor of cirrhosis progression or death and unplanned hospitalization across patients with compensated and decompensated cirrhosis. Future studies are needed to evaluate the possibility of slowing cirrhosis disease progression by reversing or preventing frailty.
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Doença Hepática Terminal , Fragilidade/diagnóstico , Cirrose Hepática , Progressão da Doença , Doença Hepática Terminal/diagnóstico , Doença Hepática Terminal/mortalidade , Doença Hepática Terminal/fisiopatologia , Feminino , Fragilidade/complicações , Fragilidade/fisiopatologia , Fragilidade/prevenção & controle , Força da Mão , Hospitalização/estatística & dados numéricos , Humanos , Cirrose Hepática/complicações , Cirrose Hepática/diagnóstico , Cirrose Hepática/fisiopatologia , Masculino , Pessoa de Meia-Idade , Mortalidade , Escores de Disfunção Orgânica , Avaliação de Resultados em Cuidados de Saúde , Equilíbrio Postural , Valor Preditivo dos Testes , Medição de Risco/métodosRESUMO
BACKGROUND AND AIMS: Sarcopenic obesity (SO) marks a confluence of 2 complex entities involving the muscle-liver-adipose tissue axis. Computed tomographic (CT) scan-derived skeletal muscle index (SMI) remains the gold standard for sarcopenia assessment in SO. However, it has intrinsic limitations of cost, radiation, and point of care applicability. We assessed the role of muscle ultrasound (US) in SO. METHODS: A total of 52 patients with cirrhosis and obesity were assessed for sarcopenia using SMI. US assessment of thigh and forearm muscles was done to record quadriceps muscle thickness (QMT), quadriceps feather index (QMFI), forearm muscle thickness (FMT), and forearm feather index (FFI), respectively. Evaluated US parameters were correlated with SMI and assessed for diagnostic accuracy using the area under the curve. RESULTS: A total of 40 (76.9%) males and 12 (23.1%) females [mean age: 50.9 y (43.8 to 53.5 y)] were included. QMT [0.45 cm/m 2 (0.42 to 0.48 cm/m 2 ) vs. 0.67 cm/m 2 (0.63 to 0.70 cm/m 2 )], QMFI [0.82 cm/m 2 (0.77 to 0.87 cm/m 2 ) vs. 1.12 cm/m 2 (1.06 to 1.19 cm/m 2 )], FMT [0.19 cm/m 2 (0.17 to 0.20 cm/m 2 ) vs. 0.25 cm/m 2 (0.23 to 0.27 cm/m 2 )], and FFI [0.38 cm/m 2 (0.35 to 0.412 cm/m 2 ) vs. 0.47 cm/m 2 (0.44 to 0.50 cm/m 2 )] were significantly lower in patients with SO ( P <0.01). A positive correlation with SMI was seen for all parameters in the entire cohort. The strongest correlation was exhibited by QMT ( r =0.70) and QMFI ( r =0.70) in males. The area under the curve of QMT, QMFI, FMT, and FFI were 0.98 (95% confidence interval: 0.96-1), 0.95 (0.89-1), 0.85 (0.75-0.96), and 0.80 (0.68-0.93), respectively. CONCLUSIONS: US-based assessment of sarcopenia has excellent diagnostic accuracy and correlates well with computed tomography-SMI in patients with SO. US may serve as an easy-to-use, point of care tool for assessing sarcopenia in SO with the advantage of repeated sequential assessment.
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Sarcopenia , Masculino , Feminino , Humanos , Pessoa de Meia-Idade , Sarcopenia/diagnóstico por imagem , Sarcopenia/etiologia , Cirrose Hepática/complicações , Cirrose Hepática/diagnóstico por imagem , Cirrose Hepática/patologia , Músculo Esquelético/diagnóstico por imagem , Músculo Esquelético/patologia , Obesidade/complicações , Obesidade/diagnóstico por imagem , Estudos RetrospectivosRESUMO
Cancer is an intricate disease that develops as a response to a combination of hereditary and environmental risk factors, which then result in a variety of changes to the genome. The cluster of differentiation (CD44) is a type of transmembrane glycoprotein that serves as a potential biomarker for cancer stem cells (CSC) and viable targets for therapeutic intervention in the context of cancer therapy. Hyaluronic acid (HA) is a linear polysaccharide that exhibits a notable affinity for the CD44 receptor. This characteristic renders it a promising candidate for therapeutic interventions aimed at selectively targeting CD44-positive cancer cells. Treating cancer via non-viral vector-based gene delivery has changed the notion of curing illness through the incorporation of therapeutic genes into the organism. The objective of this review is to provide an overview of various hyaluronic acid-modified lipoplexes and polyplexes as potential drug delivery methods for specific forms of cancer by effectively targeting CD44.
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Ácido Hialurônico , Neoplasias , Humanos , Ácido Hialurônico/metabolismo , Sistemas de Liberação de Medicamentos , Neoplasias/terapia , Linhagem Celular TumoralRESUMO
Cucumis callosus (Kachri) is an under-exploited fruit of the Cucurbitaceae family, distributed majorly in the arid regions of India in the states of Haryana, Rajasthan, and Gujarat. The fruit is traditionally used by the native people at a small scale by home-level processing. It is a perennial herb that has been shown to possess therapeutic potential in certain disorders. In several studies, the antioxidant, anti-hyperlipidaemic, anti-diabetic, anti-cancerous, anti-microbial, and cardioprotective properties of Kachri have been reported. The fruit has a good nutritional value in terms of high percentages of protein, carbohydrates, essential fatty acids, phenols, and various phytochemicals. Also, gamma radiation treatment has been used on this crop to reduce total bacterial counts (TBC), ensuring safety from pathogens during the storage period of the fruit and its products. These facts lay down a foundation for the development of functional food formulations and nutraceuticals of medicinal value from this functionally rich crop. Processing of traditionally valuable arid region foods into functional foods and products can potentially increase the livelihood and nutritional security of people globally. Therefore, this review focuses on the therapeutic and pharmacological potentials of the Kachri fruit in the management of non-communicable diseases (NCDs) namely, diabetes, cancer, and hyperlipidemia. Graphical abstract of the review.
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Cucumis , Doenças não Transmissíveis , Humanos , Doenças não Transmissíveis/tratamento farmacológico , Índia , Frutas/química , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Compostos Fitoquímicos/análiseRESUMO
Several sperm lysozyme-like genes evolved from lysozyme by successive duplications and mutations; however their functional role in the reproduction of farm animals is not well understood. To understand the function and molecular properties of buffalo sperm lysozyme-like protein 1 (buSLLP1), it was expressed in E. coli; however, it partitioned to inclusion bodies. Lowering of temperature and inducer concentration did not help in the recovery of the expressed protein in the biologically active form. Therefore, buSLLP1 was cloned and expressed in Pichiapink system based on auxotrophic Pichia pastoris in a labscale fermenter. The expressed protein was obtained in flow-through by using a 30 kDa ultrafiltration membrane followed by MonoQ anion exchange chromatography, resulting in a homogenous preparation of 40 mg recombinant buSLLP1 per liter of initial spent culture-supernatant. Circular dichroism spectroscopy showed that recombinant buSLLP1 possessed a native-like secondary structure. The recombinant buSLLP1 also showed thermal denaturation profile typical of folded globular proteins; however, the thermal stability was lower than the hen egg white lysozyme. Binding of buSLLP1 to chitin and zona pellucida of buffalo oocytes showed that the recombinant buSLLP1 possessed a competent binding pocket, therefore, the produced protein could be used to study its functional role in the reproduction of farm animals.
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Búfalos/genética , Expressão Gênica , Muramidase , Animais , Escherichia coli/genética , Escherichia coli/metabolismo , Muramidase/biossíntese , Muramidase/química , Muramidase/genética , Muramidase/isolamento & purificação , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Saccharomycetales/genética , Saccharomycetales/metabolismoRESUMO
Punicalagin is the most bioactive pomegranate polyphenol with high antioxidant and free-radical scavenging activity and can potentially cure different ailments related to the cardiovascular system. The current research work was envisioned to predict the targeting efficiency of punicalagin (PG) nanoparticles to the macrophages, more specifically to bone marrow macrophages. For this, we selected mannose-decorated PLGA-punicalagin nanoparticles (Mn-PLGA-PG), and before formulating this nanocarrier in laboratory settings, we predicted the targeting efficiency of this nanocarrier by in silico analysis. The analysis proceeded with macrophage mannose receptors to be acquainted with the binding affinity and punicalagin-based nanocarrier interactions with this receptor. In silico docking studies of macrophage mannose receptors and punicalagin showed binding interactions on its surface. PG interacted with hydrogen bonds to the charged residue ASP668 and GLY666 and polar residue GLN760 of the Mn receptor. Mannose with a docking score of -5.811 Kcal/mol interacted with four hydrogen bonds and the mannose receptor of macrophage, and in PLGA, it showed a -4.334 Kcal/mol docking score. Further, the analysis proceeded with density functional theory analysis (DFT) and HOMO-LUMO analysis, followed by an extensive 100 ns molecular dynamics simulation to analyse the trajectories showing the slightest deviation and fluctuation. While analysing the ligand and protein interaction, a wonderful interaction was found among the atoms of the ligand and protein residues. This computational study confirms that this nanocarrier could be a promising lead molecule to regulate the incidence of drug-induced neutropenia. Furthermore, experimental validation is required before this can be stated with complete confidence or before human use.
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Metotrexato , Neutropenia , Antioxidantes , Humanos , Taninos Hidrolisáveis , Ligantes , Macrófagos , Manose , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , PolifenóisRESUMO
Our study has investigated the impact of El Niño-Southern Oscillation (ENSO) on spatio-temporal dynamics of Indian summer monsoon rainfall (ISMR) as well as vegetation for a period of 1980 to 2019 at regional scale in Haryana, India. The gridded rainfall datasets of India Meteorological Department (IMD) were examined on monthly and seasonal scale using various statistical methods like mean climatology, coefficient of variation, slope of linear, Sen's slope, Mann-Kendall Z statistic, and hierarchical cluster analysis. The influence of ENSO on spatial distribution of ISMR was observed, where we found increasing and decreasing rainfall patterns during La Niña and El Niño years, respectively. We attempted to establish a link between ISMR and various teleconnections using time series of the National Oceanic and Atmospheric Administration (NOAA) Physical Sciences Laboratory, and statistically significant and positive correlation was observed with the Southern Oscillation Index (SOI), whereas significantly negative correlations were observed with SST of Niño 3, Niño 3.4, and Niño 4 regions. The gridded datasets of the European Centre for Medium-Range Weather Forecasts (ECMWF) atmospheric reanalysis V5 (ERA5) were used to evaluate the influence of ENSO on atmospheric dynamics using lower and upper tropospheric wind circulation (850 hPa and200 hpa), vertically integrated moisture transport (VIMT), and surface moisture flux (SMF). We have used satellite-based normalised difference vegetation index (NDVI) datasets of the Global Inventory Monitoring and Modeling System (GIMMS) to investigate the impact of ENSO on vegetation dynamics of Haryana and found that NDVI values were higher and lower in case of La Niña and El Niño years, respectively.
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El Niño Oscilação Sul , Monitoramento Ambiental , Monitoramento Ambiental/métodos , Índia , Estações do Ano , Tempo (Meteorologia)RESUMO
Gamma amino butyric acid (GABA) is a chemical messenger that plays a significant role in muscle relaxation and brain health. Certain lactic acid bacteria (LAB) produce significant levels of GABA and thus act as potential psychobiotic cultures. In the present study, LAB were isolated from non-rhizospheric soil sample of Syzygium cumini (Black plum). A total of 57 LAB were isolated on the basis of their morphological and acid producing characteristic on de Man Rogosa Sharpe (MRS) agar. Only seven isolates were found to produce GABA (0.09-1.13 gL-1) in MRS broth and were identified as Lactococcus. However, L. lactis LP-68 produced highest amount of GABA and was selected for further optimization of culture conditions (pH, temperature and MSG) by response surface methodology (RSM). The optimization resulted in approximately four-fold increase in GABA production (4.11 gL-1). The results indicate that the L. lactis LP-68 can be used as starter culture for production of GABA-enriched functional foods.
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Lactococcus lactis , Prunus domestica , Syzygium , Humanos , Solo , Ácido gama-AminobutíricoRESUMO
Heavy metal pollution in soil and water is a potential threat to human health as it renders food quality substandard. Different biosorbents such as microbial and agricultural biomass have been exploited for heavy metal immobilization in soil and sorptive removal in waters. Biosorption is an effective and sustainable method for heavy metal removal in soil and water, but the inherent challenges are to find cheap, selective, robust, and cost-effective bioadsorbents. Microbial and agricultural biomass and their modified forms such as nanocomposites and carbonaceous materials (viz., biochar, nanobiochar, biocarbon), might be useful for sequestration of heavy metals in soil via adsorption, ion exchange, complexation, precipitation, and enzymatic transformation mechanisms. In this review, potential biosorbents and their metal removal capacity in soil and water are discussed. The microbial adsorbents and modified composites of agricultural biomasses show improved performance, stability, reusability, and effectively immobilize heavy metals from soil and water. In the future, researchers may consider the modified composites, encapsulated biosorbents for soil and water remediation.
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Metais Pesados , Poluentes do Solo , Adsorção , Poluição Ambiental , Solo , ÁguaRESUMO
Plant pathogens resistant to the commercially available fungicides and bactericides even at higher concentrations are the biggest challenge for the farmers to control the losses due to plant diseases. The antibacterial and antifungal potential of nanomaterials makes them a suitable candidate for the control of plant diseases. Thus, the present study reports the phytofabricated zinc oxide nanoparticles (ZnO Np's) using aqueous plant leaf extract of Terminalia bellerica (Baheda). Characterization of ZnO nanoparticles was done by ultraviolet-visible (UV-Vis) studies, X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FT-IR) analysis, and transmission electron microscopy (TEM). The presence of pure hexagonal wurtzite crystalline structure of ZnO nanoparticles was confirmed by XRD analysis. The TEM images revealed the spherical to hexagonal shaped ZnO nanoparticles with sizes ranging from 20 to 30 nm. The stabilization of synthesized ZnO nanoparticles through the interactions of terpenoids, steroids, phenylpropanoids, flavonoids, phenolic acids, and enzymes present in the leaf extract was suggested by FTIR analysis. The mechanism of the formation of ZnO nanoparticles using Terminalia bellerica (Baheda) (Tb-ZnO Np's) as a bioactive compound is proposed. These phytofabricated ZnO nanoparticles (Tb-ZnO Np's) have shown significant antifungal potential against Alternaria brassicae the causal agent of Alternaria blight disease/leaf spot disease in Brassica species. The microscopic results confirm the changes in mycelium morphology and reduction in the number of spore germination at 0.2 mg/mL concentration Tb-ZnO Np's.
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Brassica , Nanopartículas Metálicas , Nanopartículas , Óxido de Zinco , Alternaria , Antibacterianos/química , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , Nanopartículas/química , Extratos Vegetais/química , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X , Óxido de Zinco/química , Óxido de Zinco/farmacologiaRESUMO
OBJECTIVE: In this research work, we hypothesized to predict the nanoparticulate system, best suited for targeted delivery of filgrastim. Significance: Targeted delivery of filgrastim to bone marrow is required to decrease the incidence of neutropenia/febrile neutropenia. This is achieved by nanoparticulate systems, duly designed by bioinformatics approach. METHOD: The targeted delivery of filgrastim in nanoparticulate system was achieved by molecular dynamics (MD) simulation studies. Two matrices comprising PLGA and SLN (tripalmitin, core component of SLN system) were modeled separately with proposed drug filgrastim. Energy minimization of all systems was done using the steepest descent method. PLGA and tripalmitin systems were equalized at 310 °C, at 1 bar pressure with Berendsen barostat for 200 ps using a v-rescale thermostat for 100 ps. Atomistic MD simulations of four model system and mass density of interacting systems were calculated. RESULTS: The mass density maps of each nanoparticle system, that is, PLGA and tripalmitin showed an increase in density toward the end of the simulation. The contact numbers attained equilibria with the average number of approx.. 1500 contacts in case of tripalmitin-filgrastim system. While PLGA-filgrastim system shows lesser contacts as compared to tripalmitin with average contacts of approx. 1000.The binding free energy was predicted to be -1104 kJ/mol in tripalmitin-filgrastim complex and -421 kJ/mol in PLGA-filgrastim system. CONCLUSION: Findings of study revealed that both nanoparticle systems assumed to be good model for drug-carrier systems. Though SLN systems were thought to be more appropriate than PLGA, still the in vivo findings could ascertain this hypothesis in futuristic work.
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Biologia Computacional , Filgrastim/química , Nanopartículas , Proteínas Recombinantes/química , Portadores de Fármacos , Fator Estimulador de Colônias de Granulócitos/químicaRESUMO
Investigations were made on the changes in physical and biochemical attributes, fruit decay and storage life of 'Winter Dawn' strawberry fruits in response to aqueous ozone dip treatment for different exposure times. Fruits were subjected to 0.1 ppm aqueous ozone for different time intervals (1-4 min). The treated strawberries were air dried and stored under ambient (25 ± 2 °C and 45-50% RH) and low temperature (2 ± 1 °C and 90% RH) conditions. Results revealed that treatment of strawberry fruits with aqueous ozone @ 0.1 ppm for 2 min resulted in 21% lower weight loss, 16% higher firmness and 15% lesser change in fruit colour during 2 days in ambient storage. Under low temperature storage, 2 min ozone treated fruits exhibited ~ 21% lower PLW, 19% higher firmness and 46% lesser colour change as compared to control fruits during 14 days of storage. Fruit decay reduced significantly under both low and cold storage conditions. Thus, it can be concluded that application of aqueous ozone for 2 min was able to retain the strawberry fruit quality and extend its storage life till 14 days under low temperature storage and 2 days under ambient storage conditions.
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MGP-40 is a mammary gland-specific glycoprotein which is expressed during involution and is an important marker for mammary gland apoptosis. It is an inactive chitinase-like protein belonging to Glycosyl Hydrolase family 18. The present study reports sequence characterization, tissue-specific expression analysis, production of recombinant MGP-40 and its mutant (A117D and L119E) in both E. coli and COS1 cells for their chitin-binding and chitinase activity analysis. The cDNA of buffalo MGP-40 was cloned and sequenced which corresponded to 1803 bp with an open reading frame of 1152 bp (361 aa), signal sequence of 63 bp (21 aa), 5' and 3' UTR of 144 bp and 507 bp, respectively. The 3' UTR analysis revealed potential sites for high level expression and stability during involution. The half-life of buffalo MGP-40 was found to be 11.7 h. MGP-40 was highly expressed in mammary gland followed by small intestine, spleen and mammary epithelial cells. The purified recombinant MGP-40 and its mutant expressed in E.coli were observed to bind chitin efficiently, however, no chitinase activity was observed. Further, chitinase activity was also not observed by expressing mutant recombinant MGP-40 in COS1 cells ruling out the possible role of post-translational modifications. Structure-based in-silico mutagenesis by FoldX algorithm showed a drastic decrease in overall fold stability which might be a possible reason for inability to recover its activity. Therefore, chitinase activity could not be restored in MGP-40 even after reverting back two critical residues in active site which may be due to detrimental effect of mutations on structural stability.
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Búfalos/metabolismo , Proteína 1 Semelhante à Quitinase-3/metabolismo , Proteína 1 Semelhante à Quitinase-3/fisiologia , Sequência de Aminoácidos , Animais , Apoptose/fisiologia , Búfalos/genética , Búfalos/fisiologia , Células COS , Proteína 1 Semelhante à Quitinase-3/genética , Quitinases/genética , Quitinases/metabolismo , Chlorocebus aethiops , Clonagem Molecular/métodos , DNA Complementar/genética , Escherichia coli/genética , Feminino , Glicoproteínas/genética , Glândulas Mamárias Animais/enzimologia , Glândulas Mamárias Animais/metabolismo , Glândulas Mamárias Animais/fisiologia , Fases de Leitura Aberta , Sinais Direcionadores de Proteínas , Proteínas Recombinantes/genéticaRESUMO
Lactoferrin (Lf) has been involved in diverse type of cellular activities and its biochemical properties are species specific. Lf is a bilobal molecule in which each lobe binds with one Fe2+/Fe3+ ion. A lot of physiological effects of Lf are regulated by its iron binding and release properties; however these properties are species-specific. To understand the iron-binding, thermal stability and cytotoxic effect of buffalo Lf (buLf) and contribution of individual N- and C-terminal lobes therein, buLf and the truncated monoferric lobes were expressed in Kluyveromyces lactis or Pichia pastoris yeast expression systems. The iron-uptake/release behavior and thermal stability of recombinant buLf was observed similar to the Lf purified from buffalo milk. Supplementation of recombinant buLf to the buffalo mammary epithelial cells (BuMEC) culture decreased their proliferation and the cell viability in a dose dependent manner. The cell growth decreased by 37% at 1.0 mg/ml Lf. C-lobe decreased the viability of BuMEC by 15% at 1 mg/ml. The C-lobe showed greater cytotoxic effect against BuMEC in comparison to N-lobe. buLf caused a reduced expression of the casein in BuMEC. At 1.0 mg/ml of buLf, CSN2 transcript level was reduced by 74% and 78% in the normal and hormone free media, respectively. The expression of IL-1ß gene in BuMEC increased by 4-5 fold in the presence of 1.0 mg/ml of Lf. The effect was similar to that observed in the involutory mammary gland, suggesting the role of elevated level of Lf in remodeling of buffalo mammary tissue during involution.
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Células Epiteliais/efeitos dos fármacos , Ferro/metabolismo , Lactoferrina/farmacologia , Animais , Búfalos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Epiteliais/citologia , Humanos , Glândulas Mamárias Humanas/citologiaRESUMO
Twelve actinobacterial strains were isolated from tomato rhizospheric soil from Manipur, a state in North East Indian Himalayan Region and screened for keratinolytic and plant growth promoting traits. Nine promising isolates were identified as Streptomyces species using partial 16S rRNA gene sequencing. Among the seven isolates showing chicken feather degradation activity, three keratinolytic strains RCM-SSR-2, -6, and -12 were found to be the most efficient feather degrading strains achieving 90% feather weight loss within 48 h of incubation. They also showed maximum keratinase and soluble peptide production. Strain RCM-SSR-2, -5, -6, -8, and -11 showed positive results for all plant growth promoting traits tested. Maximum indole-3-acetic acid production was exhibited by RCM-SSR-6. Strain RCM-SSR-1, -2, -5, -6, -9, and -11 showed antagonistic activity against three important plant pathogens. Feather hydrolysate of RCM-SSR-6 was also evaluated for in vitro seed germination test using garden pea seeds. Higher concentration of feather protein hydrolysate (3 mg ml-1 ) inhibited shoot and root length of the germinating embryo. However, lower concentration (0.01 mg ml-1 ) of feather protein hydrolysate promoted seed germination. Among the 12 strains, four isolates namely RCM-SSR-1, -2, -5, and -6 were found to be promising as multi-traits plant growth promoting rhizobacteria for development of organic fertilizer, phytostimulator, and biocontrol agents.