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Objective: The aim of study's goal was to look into the anticancer efficacy of a methanolic extract of Justicia gendarussa against a lung cancer cell line. Materials and Methods: Cell viability assays and cell and nuclear morphology examinations were used to evaluate the anticancer efficacy against methanolic extract of Justicia gendarussa on lung cancer cell lines. The IC50 doses were calculated using different concentrations of Justicia gendarussa extract (0, 10, 20, 40, 60, and 80 µg/mL). Results: The results of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay revealed that the percentage of viability in treated cells was significantly lower as compared with untreated control groups, which represented as 100%, and an inhibitory concentration of 40 µg/mL was observed. Under a phase-contrast microscope, morphological changes revealed cell shrinkage and cytoplasmic membrane blebbing. The apoptotic nuclei (intensely colored, broken nuclei, and compacted chromatin) were examined under a fluorescence microscope. Conclusions: The outcome of the research work on Justicia gendarussa was investigated for anticancer properties. The results revealed the proapoptotic and cytotoxic effects of Justicia gendarussa extract on lung cancer cell lines. From the above results and findings, it could be concluded that the Justicia gendarussa methanolic leaf extract exhibited potent anticancer activity against a lung cancer cell line. Further study needs to be conducted to investigate the active chemicals in the extract as well as the molecular mechanisms underlying its anticancer benefits.
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Background: Cancer rates continue to climb, owing largely to the world population's aging and growth, as well as economically developing countries, a surge in cancer-causing behavior, particularly smoking. The third or fourth most prevalent type of cancer is colon cancer. Cancer of the large intestine (colon) is one of the primary causes of death from cancer. Colorectal cancer prevention is mostly based on adenomatous disease screening approaches. The cytotoxic and pharmacological properties of Phoenix pusilla are widely documented. As a result, there is little recorded evidence of its cytotoxic activity against colon cancer cells. Therefore, we planned to study the efficacy of a methanolic leaf extract of Phoenix pusilla against in vitro colon cancer cells. Aim: To evaluate the anti-cancer effects of the methanolic leaf extract of Phoenix pusilla on colon cancer cell lines. Materials and Methods: In vitro screening and anti-cancer effects of the methanolic effect of Phoenix pusilla on colon cancer cell lines were assessed by cell viability assays and cell and nuclear morphological studies. For the in vitro cell culture study, different concentrations of Phoenix pusilla leaf extract (0, 25, 50, 75, 100, 150 µg/ml) were used, and IC50 doses were calculated. Results: The results of the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that the fraction of viability cells significantly decreased in treated cells when compared to untreated control groups, was expressed as 100%, and an inhibitory concentration of µg/ml was identified. A phase-contrast microscope was used to observe cell shrinkage and cytoplasmic membrane blebbing. A fluorescent microscope was used to examine the apoptotic nuclei (internally dyed nuclei, shattered nuclei, and condensed chromatin). Conclusion: In conclusion, the present study results showed that the leaf extracts of Phoenix pusilla had a strong cytotoxic effect and induced significant apoptosis in the colon cancer cell lines at a concentration of 75 µg/ml in the 24 h incubation period. More research is needed to investigate the extract's active components as well as the molecular mechanisms underlying its anti-cancer properties.
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In this paper, we fabricated poly(3,4-ethylenedioxythiophene) (PEDOT)-graphene oxide-polyphenol oxidase (PEDOT-GO-PPO) as a dopamine sensor. The morphology of PEDOT-GO-PPO was observed using scanning electron microscopy. Cyclic voltammetry was conducted to study the oxidation-reduction characteristics of dopamine. To optimize the pH, potential and limit of detection of dopamine, the amperometric technique was employed. The found limit of detection was 8 × 10-9 M, and the linear range was from 5 × 10-8 to 8.5 × 10-5 M. The Michaelis-Menten constant (K m) was calculated to be 70.34 µM, and the activation energy of the prepared electrode was 32.75 kJ mol-1. The electrode shows no significant change in the interference study. The modified electrode retains up to 80% of its original activity after 2 months. In the future, the biosensor can be used for the quantification of dopamine in human urine samples. The present modified electrode constitutes a tool for the electrochemical analysis of dopamine.
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Prophenoloxidase (proPO) is very important to protect the invertebrates from microbial infections. Our previous studies revealed that proPO was up-regulated in WSSV-injected Macrobrachium rosenbergii and is responsible for protecting M. rosenbergii from WSSV. In order to prove this mechanism, an attempt was made in the present study to silence the proPO gene in freshwater prawn by injection of dsRNA-proPO followed by WSSV challenge. Two partial fragments of proPO with the size of 251 and 331 bp were used to synthesize dsRNA using LITMUS38i vector and E. coli. The bacterially synthesized dsRNA-proPO was used to silence proPO gene to determine its involvement in developing resistance in prawn against WSSV. In proPO gene-silenced prawn, 100% mortality was observed after WSSV challenge whereas no mortality was observed in prawn injected with WSSV alone. The WSSV infection in gene-silenced prawn was confirmed by PCR, and its propagation was quantified by ELISA and real-time PCR at different time intervals. Real-time PCR assay revealed a significant reduction in the expression of proPO gene in WSSV-challenged proPO-silenced prawn when compared to normal prawn. Level of proPO was reduced significantly in the haemolymph of proPO-silenced prawn when compared to prawn injected with PBS.
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Proteínas de Artrópodes/genética , Catecol Oxidase/genética , Precursores Enzimáticos/genética , Inativação Gênica , Palaemonidae/virologia , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Proteínas de Artrópodes/metabolismo , Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Palaemonidae/enzimologia , Palaemonidae/genéticaRESUMO
The conventional image segmentation techniques have a lot of issues with highest computational cost and low level accuracy for medical image diagnosis and genome analysis. The deep learning based optimization models utilize to predict the liver cancer with RNA genome using CT images and the prediction of genome classification with NGS is a higher probable in recent medical disease classification. This paper proposes a hybrid deep learning technique constructs with SegNet, MultiResUNet, and Krill Herd optimization (KHO) algorithm to perform the extraction of the liver lesions and RNA sequencing that the optimization techniques used into the deep learning method. The proposed technique implements the SegNet for segregating the liver with genome from the CT scan; the MultiResUNet is constructed to perform the extractions of liver lesions. The KHO algorithm is combined with the deep learning approaches for tuning the hyper parameters to every Convolutional neural network model and enhances the segmentation process which may elaborately identifies the sequence that causes the liver classification disease. The proposed technique is compared with the related techniques on liver lesion classification (LL) for NGS in genome. The performance results show that the proposed technique is better to other algorithms on various performance metrics.
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Aprendizado Profundo , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/genética , Algoritmos , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Processamento de Imagem Assistida por Computador , Hepatopatias/diagnóstico por imagem , Hepatopatias/genética , Análise de Sequência de RNA , Tomografia Computadorizada por Raios XRESUMO
Cervical cancer is the fourth most communal malignant disease amongst women worldwide. In maximum circumstances, cervical cancer indications are not perceptible at its initial stages. There are a proportion of features that intensify the threat of emerging cervical cancer like human papilloma virus, sexual transmitted diseases, and smoking. Ascertaining those features and constructing a classification model to categorize, if the cases are cervical cancer or not is an existing challenging research. This learning intentions at using cervical cancer risk features to build classification model using Random Forest (RF) classification technique with the synthetic minority oversampling technique (SMOTE) and two feature reduction techniques recursive feature elimination and principle component analysis (PCA). Utmost medical data sets are frequently imbalanced since the number of patients is considerably fewer than the number of non-patients. For the imbalance of the used data set, SMOTE is cast-off to solve this problem. The data set comprises of 32 risk factors and four objective variables: Hinselmann, Schiller, Cytology and Biopsy. Accuracy, Sensitivity, Specificity, PPA and NPA of the four variables remains accurate after SMOTE when compared with values obtained before SMOTE. An RSOnto ontology has been created to visualize the progress in classification performance.
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Máquina de Vetores de Suporte , Neoplasias do Colo do Útero/epidemiologia , Fatores Etários , Algoritmos , Feminino , Contracepção Hormonal/estatística & dados numéricos , Humanos , Aprendizado de Máquina , Análise de Componente Principal , Fatores de Risco , Sensibilidade e Especificidade , Comportamento Sexual , Infecções Sexualmente Transmissíveis/epidemiologia , Fumar/epidemiologiaRESUMO
White leg shrimp, Penaeus vannamei, were collected on a monthly basis from grow-out ponds located at Tamil Nadu and Andhra Pradesh states along the east coast of India for screening of viral and other pathogens. Totally 240 shrimp samples randomly collected from 92 farms were screened for white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV), infectious myonecrosis virus (IMNV) and Enterocytozoon hepatopenaei (EHP). The number of shrimp collected from shrimp farms ranged from 6 to 20 based on the body weight of the shrimp. All the shrimp collected from one farm were pooled together for screening for pathogens by PCR assay. Among the samples screened, 28 samples were WSSV-positive, one positive for IHHNV and 30 samples positive for EHP. Among the positive samples, four samples were found to be positive for both WSSV and EHP, which indicated that the shrimp had multiple infections with WSSV and EHP. This is the first report on the occurrence of multiple infections caused by WSSV and EHP. Multiplex PCR (m-PCR) protocol was standardized to detect both pathogens simultaneously in single reaction instead of carrying out separate PCR for both pathogens. Using m-PCR assay, naturally infected shrimp samples collected from field showed two prominent bands of 615 and 510 bp for WSSV and EHP, respectively.
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Densovirinae/isolamento & purificação , Enterocytozoon/isolamento & purificação , Penaeidae/microbiologia , Penaeidae/virologia , Vírus da Síndrome da Mancha Branca 1/isolamento & purificação , Animais , Aquicultura , Coinfecção , Infecções por Vírus de DNA , Índia , Microsporidiose , Reação em Cadeia da Polimerase Multiplex/métodosRESUMO
The VP28 gene of white spot syndrome virus was amplified by PCR using gene specific primer set and cloned into pRSET B vector to produce recombinant VP28 (r-VP28) in E. coli GJ1158. The chitosan tripolyphosphate nanoparticles (CS/TPP) were prepared by ionic gelation process and characterized. The purified r-VP28 protein was encapsulated by CS/TPP nanoparticles. The encapsulation efficiency of CS/TPP nanoparticles was found to be 84.8% for r-VP28 protein binding with CS/TPP nanoparticles. The in vitro release profile of encapsulated r-VP28 was determined after treating with protease and chitosanase. The different types of feed were formulated and named as normal feed with PBS, Feed A coated with crude r-VP28, Feed B with purified r-VP28 and Feed C with CS/TPP encapsulated r-VP28 (Purified). Tissue distribution and clearance of r-VP28 at different time intervals were examined in shrimp fed with different types of feed by ELISA and the results showed the presence of r-VP28 protein in different organs. Various immunological parameters were assessed in experimental shrimp. The mRNA expression of five immune-related genes was analysed by qPCR in order to investigate their response to all types of feed in shrimp. A cumulative percentage mortality was also recorded in treated shrimp challenged with WSSV.
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Quitosana/química , Nanopartículas/química , Proteínas do Envelope Viral/genética , Vírus da Síndrome da Mancha Branca 1/genética , Animais , Quitosana/farmacologia , Escherichia coli/genética , Géis/química , Penaeidae/genética , Penaeidae/virologia , Proteínas Recombinantes/genética , Proteínas do Envelope Viral/química , Vírus da Síndrome da Mancha Branca 1/patogenicidadeRESUMO
In recent years, researchers have focused on viral and plant immunostimulants which could have beneficial effects in disease prevention and control in shrimp culture. At present, the application of the recombinant VP28 protein (r-VP28) and herbal immunostimulant has been considered as a more effective approach to prevent white spot syndrome (WSS) by enhancing the immune response in shrimp. In the present study, expression of selected immune related genes in response to r-VP28 and herbal immunostimulant mix (HIM) were separately studied qualitatively and quantitatively by RT-PCR and real time PCR, respectively during ontogenetic development from nauplius to juvenile stage in Litopenaeus vannamei. The mRNA expression level of immune related genes such as anti-lipopolysaccharides (ALF), Lysozyme, cMnSOD, Crustin, Prophenoloxidase, Tumor necrosis factor receptor-associated factor 6 (TRAF6) and Haemocyanin were found to be up-regulated significantly in different ontogenetic development stages of shrimp fed with r-VP28 and HIM formulated diets. Relative percent survival (RPS) was determined in shrimp fed with immunostimulants formulated diets after oral challenge with WSSV. The survival of WSSV challenged shrimp was found to be higher in immunostimulants treated groups when compared to untreated group. The results of PCR, ELISA and real time PCR revealed the absence of WSSV in WSSV-challenged shrimp after 20 days of treatment with immunostimulants. Among these immunostimulants, HIM was found to be more effective when compared to r-VP28. After a survey of literature, we are of the opinion that this might be the first report on the expression of immune genes during ontogenetic development of L. vannamei in response to immunostimulants.
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Adjuvantes Imunológicos/administração & dosagem , Imunidade/genética , Penaeidae/genética , Penaeidae/imunologia , Vírus da Síndrome da Mancha Branca 1/imunologia , Animais , Expressão Gênica/imunologia , Ontologia Genética , Penaeidae/virologia , RNA Mensageiro/imunologia , Proteínas Recombinantes/imunologia , Proteínas do Envelope Viral/imunologiaRESUMO
A halotolerant fungal isolate Aspergillus tubingensis GR1 was isolated from the man-made solar saltern located at Khambhat, Gujarat, India, and identified using 28S rDNA partial genome sequencing. This isolate was studied for ß-galactosidase production under solid state fermentation using wheat bran and deproteinized acid cheese whey. The influence of various agro-industrial wastes, nitrogen source and other growth conditions on ß-galactosidase production was investigated using 'one-factor-at-a-time' approach. Among various variables screened along with wheat bran and deproteinized acid cheese whey as major growth substrate, corn steep liquor and MgSO4 were found to be most significant. The optimum concentrations of these significant parameters were determined employing the response surface central composite design, revealing corn steep liquor concentration (2 mL) and magnesium sulphate (50 mg) per 5 g of wheat bran and 20 mL of deproteinized acid cheese whey for highest enzyme production (15,936 U/gds). These results suggest the feasibility of industrial large-scale production of ß-galactosidase known to be valuable in whey hydrolysis and removal of galactosyl residue from polysaccharide.
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Urinary calculi constitute one of the oldest afflictions of humans as well as animals, which are occurring globally. The calculi vary in shape, size and composition, which influence their clinical course. They are usually of the mixed-type with varying percentages of the ingredients. In medical management of urinary calculi, either the nature of calculi is to be known or the exact composition of calculi is required. In the present study, two selected calculi were recovered after surgery from two different patients for detailed examination and investigated by using Fourier-Transform infrared spectroscopy (FT-IR), thermo-gravimetric analysis (TGA), powder X-ray diffraction (XRD), scanning electron microscopy and energy dispersive analysis of X-rays (EDAX) techniques. The study demonstrated that the nature of urinary calculi and presence of major phase in mixed calculi could be identified by FT-IR, TGA and powder XRD, however, the exact content of various elements could be found by EDAX only.
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Oxalato de Cálcio/química , Microscopia Eletrônica de Varredura/métodos , Espectrometria por Raios X/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Termogravimetria/métodos , Cálculos Urinários/química , Difração de Raios X/métodos , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , PósRESUMO
The indiscriminate use of pesticides and herbicides to enhance crop production has aroused great concern, because these products are likely to reach the aquatic environment, thereby posing a health concern for humans and aquatic species. Cypermethrin (CYP), a type II pyrethroid insecticide, is widely used in agriculture and for other purposes. Therefore a study was conducted for the assessment of cytotoxic, genotoxic and oxidative stress of CYP in IEG, CB, ICG, LRG and CSG cell lines at 24h exposure. The cytotoxic effect of CYP in IEG, CB, ICG, LRG and CSG cell lines was assessed using MTT, NR, AB and CB assays. Linear correlations between each EC50 values, of CYP resulting in 50% inhibition of cytotoxicity parameters after 24h exposure to CYP were calculated for IEG, CB, ICG, LRG and CSG cell lines using MTT, NR, AB and CB assays. Statistical analysis revealed good correlation with R(2)=0.90-0.939 for all combinations between endpoints employed. The percentage of DNA damage was assessed by comet assay in IEG, CB, ICG, LRG and CSG cells exposed to CYP. The results of antioxidant parameters obtained show a significant increase in lipid peroxidation (LPO) level and decreased level of GSH, SOD and CAT in IEG, CB, ICG, LRG and CSG cell lines after exposure to increasing CYP in a concentration-dependent manner. This work proves that fish cell lines could be used not only for cytotoxicity and genotoxicity studies but also for studying oxidative stress when exposed to environmental contaminants such as pesticides and other pollutants.
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Inseticidas/farmacologia , Inseticidas/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Piretrinas/farmacologia , Piretrinas/toxicidade , Animais , Catalase/metabolismo , Linhagem Celular , Ensaio Cometa , Dano ao DNA/efeitos dos fármacos , Peixes , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacosRESUMO
The cytotoxicity, genotoxicity and oxidative stress of malachite green (MG) was investigated using the fish Channa striata kidney (CSK) and Channa striata gill (CSG) cell lines. Five concentrations ranging from 0.001 to 10 µg mL(-1) were tested in three independent experiments. Cytotoxicity was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Rhodamine 123 and Alamar Blue. The mitochondrial changes and apoptosis of MG-exposed cells were observed by Rhodamine 123 and acridine orange/ethidium bromide (AO/EB) staining, respectively. In vitro potential DNA damaging effect of MG was tested using comet assay. Mitochondrial damage, apoptosis and DNA fragmentation increased in a concentration-dependent manner. Additionally, DNA electrophoretic mobility experiments were carried out to study the binding effect of MG to double-stranded DNA (dsDNA) of cells. DNA shift mobility experiments showed that MG is capable of strongly binding to linear dsDNA causing its degradation. Biochemical parameters such as lipid peroxidation (MDA), catalase (CAT) activity and reduced glutathione (GSH) levels were evaluated after exposure to MG. In CSK and CSG cell lines exposed to MG for 48 h, a significant increase in lipid peroxidation, which might be associated with decreased levels of reduced glutathione and catalase activity in these cell lines (p < 0.001), was observed.
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Brânquias/efeitos dos fármacos , Rim/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Perciformes , Corantes de Rosanilina/toxicidade , Animais , Linhagem Celular , Ensaio Cometa , Dano ao DNA , Peixes/metabolismo , Água Doce , Brânquias/metabolismo , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Oxirredução , Perciformes/metabolismoRESUMO
Extracellular glucoamylase of Colletotrichum sp. KCP1 produced through solid state fermentation was purified by two steps purification process comprising ammonium sulphate precipitation followed by gel permeation chromatography (GPC). The Recovery of glucoamylase after GPC was 50.40 % with 19.3-fold increase in specific activity. The molecular weight of enzyme was found to be 162.18 kDa by native-PAGE and was dimeric protein of two sub-units with molecular weight of 94.62 and 67.60 kDa as determined by SDS-PAGE. Activation energy for starch hydrolysis was 26.45 kJ mol(-1) while temperature quotient (Q 10 ) was found to be 1.9. The enzyme was found to be stable over wide pH range and thermally stable at 40-50 °C up to 120 min while exhibited maximum activity at 50 °C with pH 5.0. The pKa1 and pKa2 of ionisable groups of active site controlling V max were 3.5 and 6.8, respectively. V max , K m and K cat for starch hydrolysis were found to be 58.82 U ml(-1), 1.17 mg (starch) ml(-1) and 449 s(-1), respectively. Activation energy for irreversible inactivation (E a(d)) of glucoamylase was 74.85 kJ mol(-1). Thermodynamic parameters of irreversible inactivation of glucoamylase and starch hydrolysis were also determined.
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The isolated and identified triterpenoid, 1-hydroxytetratriacontane-4-one (C34H68O2), obtained from the methanolic leaf extract of Leucas aspera Linn. was explored for the first time for antisnake venom activity. The plant (L. aspera Linn.) extract significantly antagonized the spectacled cobra (Naja naja naja) venom induced lethal activity in a mouse model. It was compared with commercial antiserum obtained from King Institute of Preventive Medicine (Chennai, Tamil Nadu, India). N. naja naja venom induced a significant decrease in antioxidant superoxide dismutase, glutathione (GSH) peroxidase, catalase, reduced GSH and glutathione-S-transferase activities and increased lipid peroxidase (LPO) activity in different organs such as heart, liver, kidney and lungs. The histological changes following the antivenom treatment were also evaluated in all these organs. There were significant alterations in the histology. Triterpenoid from methanol extract of L. aspera Linn. at a dose level of 75 mg per mouse significantly attenuated (neutralized) the venom-induced antioxidant status and also the LPO activity in different organs.
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Antioxidantes/farmacologia , Venenos Elapídicos/toxicidade , Triterpenos/farmacologia , Animais , Catalase/metabolismo , Venenos Elapídicos/antagonistas & inibidores , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Dose Letal Mediana , Camundongos , Extratos Vegetais/farmacologia , Superóxido Dismutase/metabolismoRESUMO
The present study examines the use of CS/TPP nanoparticles for gene delivery in different tissues of shrimp through oral route. The viral gene of WSSV was used to construct DNA vaccines using pcDNA 3.1, a eukaryotic expression vector and the constructs were named as pVP28. The CS/TPP nanoparticles were synthesized by ionic gelation process and these particles were characterized. The structure and morphology of the nanoparticles were studied by field emission scanning electron microscopy (FE-SEM) and FTIR (Fourier Transform Infrared Spectra). The cytotoxicity of CS/TPP nanoparticles was evaluated by MTT assay using fish cell line. The expression of gene was confirmed by Immuno-dot blot, ELISA and RT-PCR analyses. The results indicate that DNA can be easily delivered into shrimp by feeding with CS/TPP nanoparticles.
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Quitosana/administração & dosagem , Crustáceos/genética , Técnicas de Transferência de Genes , Nanopartículas/administração & dosagem , Polifosfatos/administração & dosagem , Vacinas de DNA/administração & dosagem , Vacinas Virais/administração & dosagem , Administração Oral , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quitosana/toxicidade , Peixes , Microscopia Eletrônica de Varredura , Nanopartículas/toxicidade , Nanopartículas/ultraestrutura , Polifosfatos/toxicidade , Espectroscopia de Infravermelho com Transformada de Fourier , Vacinas de DNA/genética , Vacinas de DNA/toxicidade , Vacinas Virais/genética , Vacinas Virais/toxicidade , Vírus da Síndrome da Mancha Branca 1/genéticaRESUMO
Hepatopancreatic parvovirus (HPV) which causes infection in many species of penaeid shrimp is a serious viral pathogen in the young life stages of shrimp. An attempt was made to develop an in vitro system using C6/36 subclone of Aedes albopictus cell line for propagation of HPV. The results revealed that C6/36 cells were susceptible to this virus and the infected cells showed CPE in the form of vacuole formation. The results of PCR, immunocytochemistry and Western blot revealed the HPV-infection in C6/36 cell line. The RT-PCR analysis confirmed the replication of HPV in C6/36 cell line. The HPV load was quantified at different time intervals by ELISA and real time PCR, and the results showed the increase of viral load in C6/36 cell line in time course of infection. HPV propagated in C6/36 cell line was used to infect post-larvae of shrimp and the results showed that the twentieth passage of HPV propagated in C6/36 cell line caused 100% mortality in post-larvae after 6 weeks post infection (d.p.i.). The infected post-larvae showed clinical signs of reduced growth, reduced preening, muscle opacity and atrophy of hepatopancreas. The HPV-infection was confirmed by PCR. The results of the present study showed that C6/36 cell line can be used as an in vitro model for HPV replication instead of whole animal.
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Aedes , Densovirinae/fisiologia , Penaeidae/virologia , Cultura de Vírus/métodos , Animais , Linhagem Celular , Modelos Biológicos , Replicação ViralRESUMO
An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV) of shrimp in different organs of freshwater rice-field crab, Paratelphusa hydrodomous (Herbst), using bioassay, PCR, RT-PCR, ELISA, Western blot and real-time PCR analyses, and also to use this crab instead of penaeid shrimp for the large-scale production of WSSV. This crab was found to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT-PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real-time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production.
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Braquiúros/virologia , Replicação Viral , Vírus da Síndrome da Mancha Branca 1/fisiologia , Animais , Água Doce , Perfilação da Expressão Gênica , Regulação Viral da Expressão Gênica , Fatores de Tempo , Proteínas do Envelope Viral/genética , Vírus da Síndrome da Mancha Branca 1/genéticaRESUMO
PURPOSE: Combined modality treatment using multidrug chemotherapy (CTh) and radiotherapy (RT) is currently considered the standard of care in early stage Hodgkin's disease. Its role in advanced stages, however, continues to be debated. This study was aimed at evaluating the role of consolidation radiation in patients achieving a complete remission after six cycles of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) chemotherapy using event-free survival (EFS) and overall survival (OS) as primary end points. PATIENTS AND METHODS: Two hundred and fifty-one patients with Hodgkin's disease attending the lymphoma clinic at the Tata Memorial Hospital (Mumbai, India) from 1993 to 1996 received induction chemotherapy with six cycles of ABVD after initial staging evaluation. A total of 179 of 251 patients (71%) achieved a complete remission after six cycles of ABVD chemotherapy and constituted the randomized population. Patients were randomly assigned to receive either consolidation radiation or no further therapy. RESULTS: With a median follow-up of 63 months, the 8-year EFS and OS in the CTh-alone arm were 76% and 89%, respectively, as compared with 88% and 100% in the CTh+RT arm (P =.01; P =.002). Addition of RT improved EFS and OS in patients with age < 15 years (P =.02; P =.04), B symptoms (P =.03; P =.006), advanced stage (P =.03; P =.006), and bulky disease (P =.04; P =.19). CONCLUSION: Our study suggests that the addition of consolidation radiation helps improve the EFS and OS in patients achieving a complete remission after six cycles of ABVD chemotherapy, particularly in the younger age group and in patients with B symptoms and bulky and advanced disease.