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Background: This study was carried out to evaluate the effectiveness of partial and full vaccination with ChAdOx1 nCoV-19 (COVISHIELD) to prevent the development of moderate or severe illness among COVID-positive cases. Methods: This prospective cohort study was conducted among Armed Forces personnel deployed in Northern India who were found COVID positive during the study period between January and June 2021. Information about the vaccination status, age and comorbidities was collected at the time of diagnosis. Classification of COVID cases as moderate or severe was performed as per criteria given by the Government of India. Individuals were considered partially vaccinated three weeks after one dose and fully vaccinated two weeks after the second dose. Risk ratio and vaccine effectiveness (VE) to prevent moderate or severe disease among COVID cases were calculated. Results: A total of 2005 COVID-19 patients were included in our study. Partial vaccination and full vaccination with ChAdOx1 nCoV-19 offered 13% (95% credible interval (CI): -56.8%, 52.8%) and 66.6% (95% CI: 34.9%, 84.6%) protection against progression to moderate/severe illness among COVID-positive individuals. The risk of moderate-severe disease among COVID-positive cases occurring 4-11 weeks after the first dose was also lesser among those who had taken the second dose of vaccine than individuals who have been vaccinated with only one dose. Conclusion: Interval between the first and second doses of ChAdOx1 nCoV-19 vaccine should be reduced to 4-6 weeks, as partial vaccination offers lower protection against the development of moderate-severe illness after COVID infection.
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Wool is a biodegradable fiber, rich in nutrients and can be recycled in soil as a fertilizer for maximum benefits. The present study was planned with the hypothesis that waste wool could be used as a nutrient source or manure to forage crops and aim of this study was to identify practicable recycling options of sheep based wastes in agriculture. In our study, we have compared the effect of different sheep based organic wastes on soil health, crop productivity and water use. Application of waste wool in soil significantly improved the fertility status of soil, and considerable improvement was also observed in organic carbon and nitrogen, i.e. around 30.8 and 32.6% higher over control. The activities of soil enzymes were 10-30% and 3-20% higher in waste wool treatment as compared to control and sheep based manures, respectively. Application of waste wool not only improved soil health but produced 50% higher grain and dry fodder yield of barley over control. The improvement in physical properties of soil with waste wool resulted in higher water use efficiency of the system. Our study will help in distinguishing choices for safe use of organic wastes along with up gradation of soil health and crop water utilization, particularly in nutrient poor soils of arid and semi-arid region of India.
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Fertilizantes , Solo , Agricultura , Animais , Hordeum , Índia , Esterco , Nitrogênio , OvinosRESUMO
In present investigation, differential expression of transcriptome after classical swine fever (CSF) vaccination has been explored at the cellular level in crossbred and indigenous (desi) piglets. RNA Sequencing by Expectation-Maximization (RSEM) package was used to quantify gene expression from RNA Sequencing data, and differentially expressed genes (DEGs) were identified using EBSeq, DESeq2, and edgeR softwares. After analysis, 5222, 6037, and 6210 common DEGs were identified in indigenous post-vaccinated verses pre-vaccinated, crossbred post-vaccinated verses pre-vaccinated, and post-vaccinated crossbred verses indigenous pigs, respectively. Functional annotation of these DEGs showed enrichment of antigen processing-cross presentation, B cell receptor signaling, T cell receptor signaling, NF-κB signaling, and TNF signaling pathways. The interaction network among the immune genes included more number of genes with greater connectivity in vaccinated crossbred than the indigenous piglets. Higher expression of IRF3, IL1ß, TAP1, CSK, SLA2, SLADM, and NF-kB in crossbred piglets in comparison to indigenous explains the better humoral response observed in crossbred piglets. Here, we predicted that the processed CSFV antigen through the T cell receptor signaling cascade triggers the B cell receptor-signaling pathway to finally activate MAPK kinase and NF-κB signaling pathways in B cell. This activation results in expression of genes/transcription factors that lead to B cell ontogeny, auto immunity and immune response through antibody production. Further, immunologically important genes were validated by qRT-PCR.
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Peste Suína Clássica/imunologia , Imunogenicidade da Vacina/genética , Suínos/genética , Transcriptoma , Animais , Peste Suína Clássica/genética , Peste Suína Clássica/prevenção & controle , NF-kappa B/genética , Receptores de Antígenos de Linfócitos T/genética , Transdução de Sinais , Suínos/imunologia , Fator de Necrose Tumoral alfa/genética , Vacinação/veterináriaRESUMO
The effects of three climates (hot and dry, mild and humid and Alpine) and three flock densities (<100, 100-300 and >300 m2 ) on ostrich reproductive and productive traits were studied. Data were compared with the benchmark target sets by the World Ostrich Association (Ostrich benchmark Performance Targets. Version 2, May, 2008) for reproductive qualifications of ostrich. No significant difference was observed on egg production, weight, fertility, hatchability and day-old chicks weight among the three climate conditions; however, the Alpine climate had a lower performance trend. Mild and humid climates had a significant effect of age at sexual maturity for both males and females as well as on the duration of egg production season. Stocking density did not show significant difference on egg production, hatchability, age of male and female at sexual maturity and on duration of egg production season, while an area >300 m2 showed a reduction in egg weight and day-old chick weight. Further, an area <100 m2 led to a weaker ostrich fertility rate. Results showed that the ostrich would have a better performance under hot and dry and mild and humid climates as compared to Alpine climate with a stocking density of 100-300 m2 area per breeder bird. Thus, climatic intervention strategies at Alpine regions may be carried out for maintaining optimal reproductive qualification of ostrich so as to improve the productivity in this sector.
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Criação de Animais Domésticos/métodos , Clima , Fertilidade/fisiologia , Oviposição/fisiologia , Reprodução/fisiologia , Struthioniformes , Animais , Cruzamento , Feminino , Masculino , Densidade Demográfica , Estações do AnoRESUMO
Nutrient utilization, body condition and carcass traits of cull ewes were studied in three dietary regimens based on complete feed block (CFB) feeding to control (C) with rumen protected protein (RPP), CU [RPP + urea (6 g/kg)] and CUF [RPP + urea + rumen protected fat (RPF; 40 g/kg)]. The RPP component (g/kg) in C had 1% formaldehyde-treated soy flakes 50, mustard cake 50 and sesame cake 30. The mustard and sesame cakes were replaced with urea on equivalent N basis in CU and CUF. The ewes were offered ad libitum CFB composed (g/kg) of concentrate 650, roughage 300 and molasses 50. The digestibility of OM and EE was higher (p < 0.05) in CUF than in CU and C, while that of NDF and ADF was lower (p < 0.05). The CP digestibility was higher in both CU and CUF. Utilization of N and Ca was higher (p < 0.05) in CUF compared with C and CU. Urinary purine derivatives analysis and microbial N synthesis were similar in all the groups. Blood glucose concentration improved at 90 day compared with 0 day. Ruminal attributes showed a higher (p < 0.05) pH, total N, TCA precipitable N, entodiniomorphs and total ciliate population in CU and CUF than in the control. The CUF had higher (p < 0.01) DM, digestible OM and ME intake, which resulted in higher (p < 0.05) weight gain and better feed efficiency. Ewes in all the groups showed an improvement in carcass traits at 90 day. The pre-slaughter weight was higher (p < 0.05) and dissected and KOH bone content was lower in CUF. Thus, feeding of CFB with urea as a cheaper N source and RPF to enrich with energy supported the improved performance as evidenced from higher nutrient input, 'utilization and efficiency with enhanced carcass traits' for better marketability and returns from cull ewes.
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Ração Animal/análise , Composição Corporal , Proteínas Alimentares/análise , Rúmen/fisiologia , Ovinos/fisiologia , Ureia/administração & dosagem , Abate de Animais , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Gorduras na Dieta/administração & dosagem , Gorduras na Dieta/análise , Proteínas Alimentares/administração & dosagem , Fermentação , Nitrogênio/administração & dosagem , Nitrogênio/químicaRESUMO
The canine parvovirus NS1 (CPV2.NS1) protein selectively induces apoptosis in the malignant cells. However, for an effective in vivo tumor treatment strategy, an oncolytic agent also needs to induce a potent anti-tumor immune response. In the present study, we used poly (I:C), a TLR3 ligand, as an adjuvant along with CPV2.NS1 to find out if the combination can enhance the oncolytic activity by inducing a potent anti-tumor immune response. The 4T1 mammary carcinoma cells were used to induce mammary tumor in Balb/c mice. The results suggested that poly (I:C), when given along with CPV2.NS1, not only significantly reduced the tumor growth but also augmented the immune response against tumor antigen(s) as indicated by the increase in blood CD4+ and CD8+ counts and infiltration of immune cells in the tumor tissue. Further, blood serum analysis of the cytokines revealed that Th1 cytokines (IFN-γ and IL-2) were significantly upregulated in the treatment group indicating activation of cell-mediated immune response. The present study reports the efficacy of CPV2.NS1 along with poly (I:C) not only in inhibiting the mammary tumor growth but also in generating an active anti-tumor immune response without any visible toxicity. The results of our study may help in developing CPV2.NS1 and poly (I: C) combination as a cancer therapeutic regime to treat various malignancies.
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Antineoplásicos/farmacologia , Neoplasias Mamárias Experimentais/tratamento farmacológico , Parvovirus Canino/química , Poli I-C/farmacologia , Proteínas não Estruturais Virais/farmacologia , Animais , Apoptose , Citocinas/sangue , Feminino , Humanos , Neoplasias Mamárias Experimentais/imunologia , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Neovascularização Fisiológica , Linfócitos T Citotóxicos/imunologia , Células Th1/imunologiaRESUMO
BACKGROUND AND PURPOSE: The role of inheritance in ascertaining susceptibility to epilepsy is well established, although the pathogenetic mechanisms are still not very clear. Interviewing for a positive family history is a popular epidemiological tool in the understanding of this susceptibility. Our aim was to visualize and localize network abnormalities that could be associated with a positive family history in a group of patients with hot water epilepsy (HWE) using resting-state functional magnetic resonance imaging (rsfMRI). METHODS: Graph theory analysis of rsfMRI (clustering coefficient γ; path length λ; small worldness σ) in probands with a positive family history of epilepsy (FHE+, 25) were compared with probands without FHE (FHE-, 33). Whether a closer biological relationship was associated with a higher likelihood of network abnormalities was also ascertained. RESULTS: A positive family history of epilepsy had decreased γ, increased λ and decreased σ in bilateral temporofrontal regions compared to FHE- (false discovery rate corrected P ≤ 0.0062). These changes were more pronounced in probands having first degree relatives and siblings with epilepsy. Probands with multiple types of epilepsy in the family showed decreased σ in comparison to only HWE in the family. CONCLUSION: Graph theory analysis of the rsfMRI can be used to understand the neurobiology of diseases like genetic susceptibility in HWE. Reduced small worldness, proportional to the degree of relationship, is consistent with the current understanding that disease severity is higher in closer biological relations.
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Encéfalo/diagnóstico por imagem , Epilepsia/diagnóstico por imagem , Rede Nervosa/diagnóstico por imagem , Adolescente , Adulto , Criança , Conectoma , Família , Feminino , Neuroimagem Funcional , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Irmãos , Adulto JovemRESUMO
Hemagglutinin neuraminidase (HN) is a membrane protein of Newcastle disease virus (NDV) with the ability to induce apoptosis in many transformed cell lines. TNF-α is a multi-factorial protein that regulates cell survival, differentiation and apoptosis. In a previous study, we reported that HN protein induces apoptosis by downregulating NF-κB expression. Further, we speculated that downregulation of NF-κB expression might sensitize HeLa cells to TNF-α-mediated apoptosis. Therefore, the present study was undertaken to investigate if HN protein could sensitize HeLa cells to TNF-α and to examine the apoptotic potential of the HN protein and TNF-α in combination. The results revealed that the pro-apoptotic effects were more pronounced with the combination of HN and TNF-α than with HN or TNF-α alone, which indicates that the HN protein indeed sensitized the HeLa cells to TNF-α-induced cell death. The results of the study provide a mechanistic insight into the apoptotic action of HN protein along with TNF-α, which could be valuable in treating tumor types that are naturally resistant to TNF-α.
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Apoptose/fisiologia , Proteína HN/metabolismo , NF-kappa B/metabolismo , Vírus da Doença de Newcastle/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Apoptose/efeitos dos fármacos , Regulação para Baixo , Regulação da Expressão Gênica , Proteína HN/genética , Células HeLa , Humanos , NF-kappa B/genética , Regulação para Cima , Proteínas Virais/metabolismo , Proteínas Virais/farmacologiaRESUMO
Many viruses have the ability to modulate the apoptosis, and to accomplish it; viruses encode proteins which specifically interact with the cellular signaling pathways. While some viruses encode proteins, which inhibit the apoptosis or death of the infected cells, there are viruses whose encoded proteins can kill the infected cells by multiple mechanisms, including apoptosis. A particular class of these viruses has specific gene(s) in their genomes which, upon ectopic expression, can kill the tumor cells selectively without affecting the normal cells. These genes and their encoded products have demonstrated great potential to be developed as novel anticancer therapeutic agents which can specifically target and kill the cancer cells leaving the normal cells unharmed. In this review, we will discuss about the viral genes having specific cancer cell killing properties, what is known about their functioning, signaling pathways and their therapeutic applications as anticancer agents.
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Genes Virais , Neoplasias/terapia , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/genética , Adenoviridae/genética , Animais , Apoptose , Proteínas do Capsídeo/análise , Proteínas do Capsídeo/genética , Vírus da Anemia da Galinha/genética , Humanos , Parvovirus/genéticaRESUMO
Thirty-six Malpura lambs (28 day old and 6.7 ± 0.25 kg BW) were distributed equally in three groups having six males and six female. They were ad libitum fed individually three different experimental diets containing calcium soap of fatty acids (CA-FA) at 0 (T1 ) and 40 (T2 and T3 ) g/kg concentrate up to six months of age. Animals in T3 were supplemented additionally with 40 mg DL-α-tocopherol acetate/kg of concentrate. The roughage moiety included ad libitum dry Prosopis cineraria and fresh Azadirachata indica leaves. All the lambs were allowed to suckle from their dam up to weaning (90 day of age). Supplementation of Ca-FA improved weight gain and feed conversion ratio during both pre- (28-90 days) and post-weaning (91-180 days) phases; however, no effect of DL-α-tocopherol was observed. Metabolic parameters during post-weaning phase revealed non-significant effect on digestibility but improved nitrogen balance in the test groups. The effect on biochemical attributes did not show any significant alteration in ruminal parameters, blood biochemicals and urinary purine derivatives. Carcass traits revealed higher (p < 0.05) dressing yield and loin eye area with Ca-FA supplementation. The value of thiobarbituric reactive substances for nuggets prepared from frozen carcasses revealed significant (p < 0.05) reduction in T3 compared to the other dietary groups. Fatty acid profile of adipose tissue revealed higher (p < 0.001) 9-octadecanoic, 9-12-octadecadienoic, polyunsaturated fatty acids (PUFA), higher ratio of PUFA/saturated fatty acids (SFA), ω-6/ω-3 and lower SFA in Ca-FA-supplemented groups. It is concluded that supplementation of 40 g/kg calcium soap prepared from industrial grade rice bran oil in lamb ration provided additional energy intake, improved N utilization, gain and feed conversion ratio besides improving dressing yield and meat quality with CLA enriched fatty acid profile. DL-α-tocopherol acetate when supplemented at 40 mg/kg feed reduced lipid oxidation of meat products thus improving its keeping quality.
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Cálcio/química , Dieta/veterinária , Ácidos Graxos/farmacologia , Carne/normas , Óleos de Plantas/química , alfa-Tocoferol/farmacologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Suplementos Nutricionais , Ácidos Graxos/administração & dosagem , Ácidos Graxos/química , Feminino , Masculino , Óleo de Farelo de Arroz , Rúmen/fisiologia , Ovinos/crescimento & desenvolvimento , alfa-Tocoferol/administração & dosagemRESUMO
Newcastle Disease (ND) is one of the major causes of economic loss in the poultry industry. Newcastle Disease Virus (NDV) is a single-stranded, negative-sense enveloped RNA virus (Fam. Paramyxoviridae; Order Mononegavirales). In the present study three monoclonal antibodies (MAbs) were produced by polyethyleneglycol (PEG)-mediated fusion of lymphocytes sensitized to NDV Bareilly strain and myeloma cells. NDV possesses ability to agglutinate erythrocytes of avian species. All the three MAbs designated as 2H7, 3E9 and 3G6 caused hemagglutination inhibition of NDV by specifically binding to NDV. The reactivity for all the 3 MAbs on indirect ELISA was found to be significantly higher than the antibody and antigen controls. On flowcytometry of HeLa cells infected with NDV using the MAbs as primary antibodies, there was a significant difference in the percentage of cells showing positive fluorescence compared to the mock control. One of the MAbs (3E9) was found to react with hemagglutinin-neuraminidase (HN) protein on western blot.
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Anticorpos Monoclonais/biossíntese , Anticorpos Antivirais/biossíntese , Vírus da Doença de Newcastle/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Ensaio de Imunoadsorção Enzimática , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência MolecularRESUMO
Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.
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Apoptose/genética , Proteínas do Capsídeo/genética , Vírus Oncogênicos/genética , Terapia Viral Oncolítica , Proteínas não Estruturais Virais/genética , Animais , Proteínas do Capsídeo/uso terapêutico , Galinhas/virologia , Cães , Vetores Genéticos , Células HeLa , Humanos , Parvovirus Canino/genética , Proteínas não Estruturais Virais/uso terapêuticoRESUMO
Indian shad, Tenualosa ilisha, is a commercially important anadromous fish representing major catch in Indo-pacific region. The present study evaluated partial Cytochrome b (Cyt b) gene sequence of mtDNA in T. ilisha for determining genetic variation from Bay of Bengal and Arabian Sea origins. The genomic DNA extracted from T. ilisha samples representing two distant rivers in the Indian subcontinent, the Bhagirathi (lower stretch of Ganges) and the Tapi was analyzed. Sequencing of 307 bp mtDNA Cytochrome b gene fragment revealed the presence of 5 haplotypes, with high haplotype diversity (Hd) of 0.9048 with variance 0.103 and low nucleotide diversity (π) of 0.14301. Three population specific haplotypes were observed in river Ganga and two haplotypes in river Tapi. Neighbour-joining tree based on Cytochrome b gene sequences of T. ilisha showed that population from Bay of Bengal and Arabian Sea origins belonged to two distinct clusters.
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DNA Mitocondrial/genética , Peixes/genética , Variação Genética , Animais , FilogeniaRESUMO
This study was conducted to evaluate the effect of mineral and antioxidant supplementation on growth, reproductive performance and physiological adaptability of heat-stressed Malpura ewes. The study was conducted for a period of 21 days in 21 adult Malpura ewes. The ewes were randomly divided into three groups with seven animals each viz. GI (control; n = 7), GII (heat stress; n = 7) and GIII (heat stress + mineral and antioxidant supplementation; n = 7). The animals were stall fed ad libitum with the diet consisting of 70% roughage and 30% concentrate. GI ewes were maintained under normal controlled condition in the shed, while GII and GIII ewes were subjected to heat stress by exposing them to 42 °C in the climatic chamber. The parameters studied were feed intake (FI), water intake (WI), body weight, body condition score (BCS), physiological, biochemical and endocrine responses. Heat stress significantly altered FI, water intake, BCS, respiration rate and rectal temperature in the afternoon, oestrus duration, estradiol, progesterone, Hb, PCV, plasma glucose, total protein, cortisol, T3 and T4 levels while mineral and antioxidant supplementation ameliorated this heat stress effect on the parameters studied. Further, the adverse effect of heat stress on the productive and reproductive efficiency of Malpura ewes was reduced considerably by mineral mixture and antioxidant supplementation. This is evident from the non-significant difference in BCS, oestrus duration and plasma estradiol between GI and GIII in this study. Hence, it is very pertinent to conclude from this study that mineral mixture and antioxidant supplementation were able to protect Malpura ewes against heat stress.
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Adaptação Fisiológica/efeitos dos fármacos , Antioxidantes/farmacologia , Temperatura Alta , Minerais/farmacologia , Reprodução/efeitos dos fármacos , Ovinos/fisiologia , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Antioxidantes/administração & dosagem , Dieta/veterinária , Suplementos Nutricionais , Feminino , Minerais/administração & dosagem , Estresse FisiológicoRESUMO
Frozen shrimp continued to be the single largest item of export from India in terms of value accounting for about 44% of the total marine export earnings. Headless, peeled frozen shrimp is a common and dominant item in the market and there is need for differentiating peeled Penaeus sp from Metapenaeus, Parapenopsis and Macrobrachium sp as consumer preference and price vary. Furthermore, there is need to find out original species used in value addition of shrimp products. Hence, it is essential for development of simple and consumer friendly technique for the identification of shrimp and their products in the market. Two monoclonal antibodies (MAbs) C-15 (IgG3) and C-52 (IgG2a) reacting with 65 and 47 kD proteins of Penaeus monodon respectively in the Western blot were selected. In epitope analysis by immunodot, the two MAbs reacted and recognized specific proteins of P. monodon, Fenneropenaeus indicus and Littopenaeus vannamei and not that of Metapenaeus, Parapenopsis, Macrobrachium rosenbergii, crabs and fishes. The immunodot required 120 min for completion. The sensitivity of the immunodot to detect proteins of P. monodon was 0.225 mg with MAb C-15 and 0.028 mg with MAb C-52. The MAb based immunodot developed, could be used for identifying and differentiating meat of P. monodon, F. indicus, and L. vannamei from that of Metapenaeus, Parapenopsis, M. rosenbergii, crabs and fishes.
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Carbon foot prints (CFs) studies based on life cycle assessment between sheep farming systems and green house gases (GHG) emissions is one of the best indicators to quantify the amount of GHG emissions per kg of product. Therefore, a life cycle assessment (LCA) study was conducted for three different sheep farming systems i.e. intensive system (stall fed only), semi-intensive (grazing with supplementation) and extensive system (grazing only) under semiarid region of India to assess the carbon cost of sheep rearing. The total CFs were estimated to be 16.9, 15.8 and 17.1 kg CO2-eq in intensive, semi-intensive and extensive system of grazing indicating semi-intensive system to be most carbon (C) efficient. For 1kg mutton production in semi-intensive and intensive system, around 30% and 24% CFs were contributed from enteric fermentation and feed respectively, whereas, in extensive system, the contribution of enteric fermentation increased up to 50%. The carbon foot prints analysis gives an insight of carbon inputs used but the amount of CO2 sequestered in soil making LCA a holistic approach for estimating GHG emissions from livestock.
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Pegada de Carbono , Efeito Estufa , Animais , Ovinos , Dióxido de Carbono , Indústria de Laticínios , Agricultura , Carbono , Estágios do Ciclo de VidaRESUMO
INTRODUCTION: Acellular matrices have historically been applied as biologic scaffolds in surgery, wound care, and tissue engineering, albeit with inconsistent outcomes. One aspect that varies widely between products is the selection of decellularization protocol, yet few studies assess comparative effectiveness of these protocols in preserving mechanics, and protein content. This study characterizes bladder acellular matrix (BAM) using two different detergent and enzymatic protocols, evaluating effects on nuclei and DNA removal (≥90%), structure, tensile properties, and maintenance of extracellular matrix proteins. METHODS: Porcine bladders were decellularized with 0.5% Sodium Dodecyl Sulfate (SDS) or 0.25% Trypsin-hypotonic-Triton X-100 hypertonic (TT)-based agitation protocols, followed by DNase/RNase agents. Characterization of BAM included decellularization efficacy (DAPI, DNA quantification), structure (histology and scanning electron microscopy), tensile testing (Instron 345C-1 mechanical tester), and protein presence and diversity (mass spectrometry). SDS and TT data was directly compared to the same native bladder using two-tailed paired t-tests. Native, TT, and SDS cohorts for tensile testing were compared using one-way ANOVA; Tukey's post-hoc tests for among group differences. RESULTS: Effective nuclei removal was achieved by SDS- and TT-based protocols. However, target DNA removal was achieved with SDS but not TT. SDS more effectively maintained qualitative tissue architecture compared to TT. The tensile modulus of the TT cohort increased, and stretchability decreased after decellularization in both SDS and TT. UTS was unaffected by either protocol. Higher overall diversity and quantity of core matrisome and matrisome-associated proteins was maintained in the SDS vs TT cohort post-decellularization. CONCLUSION: The results indicated that detergent selection affects multiple aspects of the resultant BAM biologic product. In the selected protocols, SDS was superior to TT efficacy, and maintenance of gross tissue architecture as well as maintenance of ECM proteins. Decellularization increased scaffold resistance to deformation in both cohorts. Future studies applying biologic scaffolds must consider the processing method and agents used to ensure that materials selected are optimized for characteristics that will facilitate effective translational use.
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Viral gene oncotherapy is emerging as a biotherapeutic cancer treatment modality based on targeted killing of cancer cells by viral genes. Newcastle disease virus (NDV) has the property to cause selective oncolysis of tumor cells sparing normal cells. NDV has a single stranded negative sense RNA genome, which is 15,186 nucleotide long and consists of six genes, which codes for eight proteins. NDV like other paramyxoviruses has the ability to generate multiple proteins from the P gene. P protein is encoded by an unedited transcript of the P gene, whereas the V and W protein are the results of RNA editing event in which one and two G residues are inserted at a conserved editing site within the P gene mRNA resulting in V and W transcripts, respectively. Although NDV is known to cause oncolysis by triggering apoptosis, the role of different viral proteins in selective oncolysis is still unclear. P gene edited products are known for its anti-apoptotic property in homologous host. In the present study, NDV P gene and its RNA edited products were amplified, cloned, sequenced and in vitro expression was done in HeLa cells. Further constructs were assayed for their apoptosis inducing ability in HeLa cells. Preliminary study suggested that P, V and W proteins are not apoptotic to HeLa cells.
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Genes Virais/genética , Vírus da Doença de Newcastle/genética , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Animais , Anexina A5/metabolismo , Sequência de Bases , Galinhas , Clonagem Molecular , Regulação Viral da Expressão Gênica , Células HeLa , Humanos , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Fosfoproteínas/química , Reprodutibilidade dos Testes , Proteínas Virais/químicaRESUMO
Forty two Malpura lambs (21 d old) were divided into three groups of 14 each consisting of 8 females and 6 males. Lambs were allowed to suckle their respective dams twice daily up to weaning (13 wks) and offered free choice concentrate and roughage in a cafeteria system. The lambs in control group were fed conventional concentrate mixture, in RBO group concentrate mixture fortified with 4% industrial grade rice bran oil and in Ca-soap rice bran oil (as in RBO group) was supplemented in the form of calcium soap. The concentrate intake decreased(p≤0.05) in RBO group as a result total dry matter, crude protein and metabolizable energy intake decreased compared to control whereas Ca-soap prepared from the same rice bran oil stimulated the concentrate intake leading to higher total dry matter, crude protein and energy intakes. The digestibility of dry matter (p≤0.05), organic matter (p≤0.05) and crude protein (p≤0.05) was higher in RBO group followed by Ca-soap and control whereas no effect was observed for ether extract digestibility. Higher cholesterol (p≤0.05) content was recorded in serum of oil supplemented groups (RBO and Ca-soap) while no effect was recorded for other blood parameters. Rice bran oil as such adversely affected and reduced the body weight gain (p≤0.001) of lambs in comparison to control whereas the Ca-soap of rice bran oil improved body weight gain and feed conversion efficiency in lambs. Fat supplementation decreased total volatile fatty acids (p≤0.05) and individual volatile fatty acid concentration which increased at 4 h post feeding. Fat supplementation also reduced (p≤0.05) total protozoa count. Ca-soap of rice bran oil improved pre slaughter weight (p≤0.05) and hot carcass weight (p≤0.05). It is concluded from the study that rice bran oil in the form of calcium soap at 40 g/kg of concentrate improved growth, feed conversion efficiency and carcass quality as compared to rice bran oil as such and control groups.