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1.
Rev. ADM ; 78(5): 275-279, sept.-oct. 2021.
Article in Spanish | LILACS | ID: biblio-1348224

ABSTRACT

El SARS-CoV-2, causante de que estemos viviendo una pandemia mundial, tuvo sus orígenes en China, desde donde ha traspasado fronteras rápidamente, llegando a todos los rincones del mundo. Muchos han sido los equipos de investigación que se enfrentan el reto de conseguir una vacuna que logre combatir este mortal virus. Es por este motivo que en esta investigación se pretendió analizar la bibliografía referida a la vacuna Johnson & Johnson (J&J) contra COVID-19: distribución mundial de la vacuna, mecanismo de acción, indicaciones, contraindicaciones y efectos secundarios. Varios estudios demuestran que su eficacia varía de acuerdo con la edad y género de cada individuo; sin embargo, esta vacuna alcanzó un grado de certeza moderada. Los efectos adversos en su mayoría son leves y se resolvieron al cabo de dos días, siendo excepción algunos casos, ya que se registró un efecto adverso poco común denominado trombocitopenia prevalente en mujeres de 18 a 40 años, por este motivo, la FDA (Administración de Alimentos y Medicamentos de EE.UU.) recomienda la precaución en el uso de la vacuna con respecto a este efecto adverso que en algunos casos podría ser mortal (AU)


The SARS-CoV-2, which caused us to be experiencing a global pandemic, had its origins in China, from where it has crossed borders rapidly, reaching all corners of the world. Many research teams have faced the challenge of getting a vaccine to fight this deadly virus. For this reason, this research aimed to analyze the literature on the Johnson & Johnson COVID-19 vaccine: global distribution of the vaccine, mechanism of action, indications, contraindications and side effects. Several studies show that its effectiveness varies according to the age and gender of each individual, but this vaccine reached a moderate degree of certainty. The adverse effects are mostly mild and resolved within two days, with some exceptions being a rare adverse effect called prevalent thrombocytopenia in women aged 18 to 40 years. For this reason, the FDA recommends caution in the use of the vaccine with respect to this potentially fatal adverse effect in some cases (AU)


Subject(s)
Humans , Male , Female , Contraindications, Drug , COVID-19 Vaccines/adverse effects , COVID-19 Vaccines/supply & distribution , COVID-19 Vaccines/therapeutic use , COVID-19 Vaccines/pharmacology , SARS-CoV-2 , COVID-19/prevention & control , United States Food and Drug Administration , Viral Proteins , Effectiveness , RNA, Viral , Sex Factors , Age Factors , Virus Inactivation
2.
rev. cuid. (Bucaramanga. 2010) ; 12(1): e1273, ene-2021.
Article in Spanish | LILACS, BDENF, COLNAL | ID: biblio-1177826

ABSTRACT

La enfermedad por coronavirus 2019 (COVID-19), una afección respiratoria aguda causada por el SARS-CoV2, ha sido clasificada como pandemia por la Organización Mundial de la Salud (OMS) una vez que se ha expandido a 215 países del mundo, ha infectado a más de 7.800.000 personas y cerca de 440.000 personas han muerto por su causa1. El SARS-CoV2 es un coronavirus tipo ß de características genómicas similares al MERS-CoV y al SARS-CoV1 los cuales afectaron a más de 10.000 personas en las últimas dos décadas2,3. Debido a la alta tasa de contagio y propagación del SARS-CoV2, diversas medidas de mitigación han sido empleadas. Entre las medidas adoptadas se encuentran la cuarentena, el distanciamiento físico, la limpieza de superficies y aerosoles, así como el uso de equipos de protección personal (EPP)2­6 Dado que no se dispone de vacuna y tratamientos suficientemente efectivos, evitar el contagio por contacto directo con superficies, aerosoles o suspensiones contaminadas con el SARS-CoV2 es la primera y más importante medida de contención contra la pandemia7. Esto obedece a que el SARS-CoV2 puede sobrevivir por 3 h en aerosoles, 4 h en superficies de cobre, un día en cartón, dos días en acero inoxidable y hasta 3 días en plástico8. Dada la alta demanda de los EPP, que provoca escasez y obliga a la reutilización de estos7,9, métodos eficaces de limpieza son cruciales.


Subject(s)
Humans , Virus Inactivation , Anti-Bacterial Agents
3.
Int. j. high dilution res ; 19(4): 2-9, 2020.
Article in English | LILACS, HomeoIndex | ID: biblio-1146533

ABSTRACT

IntroductionNosodes, the homeopathicpreparationssourcedfrom biological materials including clinical samples, cultures of organisms, and diseased tissues have been in use against the source-specific infections as well as other diseases. The nosodes have demonstrated some efficacy in managing epidemics, such as influenza, dengue, and leptospirosis.This article presents the need and process of development ofnosodes from the SARS-CoV-2 to explore its prophylactic and therapeutic potentials against certain related viral diseases.Materials and methodsA clinical sample of SARS-Cov-2 positive patient,based on the cycle threshold (CT) value of the qRT-PCR, heat-inactivated SARS-CoV-2, and spike glycoprotein all were processed for making nosodesas per the method described in Homoeopathy Pharmacopoeia of India.Molecular tests, such as qRT-PCR and sterility tests were performed to establish the live organisms, RNA material, and the absence of contamination.ResultsThree variants of CoronavirusNosodewere developed using a clinical sample,heat-inactivatedSARS-CoV-2, and spike glycoprotein.In potencies 3c and above, no detectableSARS-CoV-2 RNA material was found by PCR.The analytical results for nosodes were reported as compliant for sterility testing as per the IP.ConclusionThree variants of Coronavirus nosodes were preparedwhich need to be evaluated further through pre-clinical and clinical studies.(AU)


Subject(s)
Humans , Nosodes (Homeopathy)/pharmacology , Coronavirus Infections/therapy , Drug Compounding , Spike Glycoprotein, Coronavirus , Betacoronavirus , Virus Inactivation , Betacoronavirus/drug effects
5.
Article in English | WPRIM | ID: wpr-773410

ABSTRACT

OBJECTIVE@#To investigate the mechanisms underlying ozone-induced inactivation of poliovirus type 1 (PV1).@*METHODS@#We used cell culture, long-overlapping RT-PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations.@*RESULTS@#Our results indicated that ozone inactivated PV1 primarily by disrupting the 5'-non-coding region (5'-NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80-124 nucleotide (nt) region in the 5'-NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non-infectious.@*CONCLUSION@#In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80-124 nt region in the 5'-NCR is targeted by ozone to achieve this inactivation.


Subject(s)
5' Untranslated Regions , Animals , Chlorocebus aethiops , Genome, Viral , Oxidants, Photochemical , Pharmacology , Ozone , Pharmacology , Poliovirus , Vero Cells , Virus Inactivation
6.
Article in English | WPRIM | ID: wpr-690659

ABSTRACT

<p><b>OBJECTIVE</b>The current study aims to investigate the effect of Hemagglutinating virus of Japan envelope (HVJ-E) on induction of apoptosis and autophagy in human prostate cancer PC3 cells, and the underlying mechanisms.</p><p><b>METHODS</b>PC3 cells were treated with HVJ-E at various multiplicity of infection (MOI), and the generated reactive oxygen species (ROS), cell viability, apoptosis, and autophagy were detected, respectively. Next, the role of ROS played in the regulation of HVJ-E-induced apoptosis and autuphagy in PC3 cells were analysed. In the end, the relationship between HVJ-E-induced apoptosis and autuophagy was investigated by using rapamycin and chloroquine.</p><p><b>RESULTS</b>Flow cytometry assay revealed that HVJ-E treatment induced dose-dependent apoptosis and that the JNK and p38 MAPK signaling pathways were involved in HVJ-E-induced apoptosis in PC3 cells. In addition, HVJ-E was able to induce autophagy in PC3 cells via the class III PI3K/beclin-1 pathway. The data also implyed that HVJ-E-triggered autophagy and apoptosis were ROS dependent. When ROS was blocked with N-acetylcysteine (NAC), HVJ-E-induced LC3-II conversion and apoptosis were reversed. Interestingly, HVJ-E-induced apoptosis was significantly increased by an inducer of autophagy, rapamycin pretreatment, both in vitro and in vivo.</p><p><b>CONCLUSION</b>HVJ-E exerts anticancer effects via autophagic cell death in prostate cancer cells.</p>


Subject(s)
Apoptosis , Physiology , Autophagy , Physiology , Cell Line, Tumor , Cell Survival , Humans , Male , Oncolytic Virotherapy , Prostatic Neoplasms , Metabolism , Reactive Oxygen Species , Metabolism , Sendai virus , Allergy and Immunology , Physiology , Virus Inactivation
7.
Infectio ; 21(4): 251-254, Oct.-Dec. 2017. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-892739

ABSTRACT

Resumen Objetivo: Evaluar al método de inactivación del carbapenémico (MIC*) frente a técnicas como el Test de Hodge modificado (THM), ácido 3-aminofenilborónico (APB) y la reacción en cadena de la polimerasa en enterobacterias productoras de carbapenemasas (EPC) tipo KPC. Materiales y métodos: Se seleccionaron 88 aislados clínicos de K. pneumoniae, K. oxytoca, E.coli, S. marcescens, C. freundii sensibles y 91 resistentes a los carbapenémicos. El APB y el método MIC* se realizaron siguiendo las publicaciones originales. El THM se realizó de acuerdo al CLSI 100S Edición 26-2016. El gen blaKPC se identificó por multiplex PCR. Resultados: El MIC* en EPC tipo KPC presentó una sensibilidad/especificidad cercana al 100% y kappa de 1 comparado con la PCR; se observó la ausencia de halo en todas los aislados EPC tipo KPC a diferencia de los aislados sensibles a los cabapenémicos que presentaron halo > 19mm. Se observó el 3 % de resultados falsos positivos y el 5 % de falsos negativos en THM y ABP respectivamente. Discusión y conclusiones: El MIC* y la PCR demuestran superioridad al THM y ABP para identificar carbapenemasas tipo KPC en EPC. Se recomienda su uso de forma rutinaria dentro del algoritmo para la contención de infecciones por este tipo de patógenos.


Abstract Objective: To compare the carbapenem inactivation method (CIM *) with the Modified Hodge Test (MHT), the acid 3-aminophenylboronic test(APB) and the polymerase chain reaction (PCR) detection of the blaKPC gene for the identification of KPC carbapenemase producing Enterobacteriaceae (ECP). Materials and Methods: We selected 88 susceptible and 91 carbapenems resistant clinical isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Escherichia coli, Serratia marcescens and Citrobacter freundii. We performed APB and CIM* according to previously published methods and the MHT according to CLSI 100S Edition 26-2016. The blaKPC gene was identified by PCR multiplex. Results: The CIM* had a sensitivity and specificity close to 100% and a kappa score of 1 compared with gold standard PCR. The absence of zone diameter was observed in all isolated KPC producers, unlike in isolates susceptible to carbapenems, where a zone diameter >19mm was observed. Three percent of false positive and five percent of false negative was observed in THM and ABP respectively. Discussion and conclusions: The CIM* and the PCR were better than MHT and ABP at identifying carbapenemases in ECP. We recommend the routine use of the CIM* within the algorithm for ECP infection control.


Subject(s)
Humans , Carbapenem-Resistant Enterobacteriaceae , Polymerase Chain Reaction , Low Cost Technology , Virus Inactivation , Enterobacteriaceae
8.
Iranian Journal of Veterinary Research. 2017; 18 (1): 6-12
in English | IMEMR | ID: emr-189260

ABSTRACT

Reoviruses are important pathogens responsible for poor growth performance and silent losses in the poultry industry. They are associated with many disease and syndromes such as malabsorption [runting and stunting syndrome], respiratory diseases and immunosuppression. Broiler birds are most susceptible to viral infections during the early post hatching period. Therefore, the transfer of maternal immunity to embryonated eggs is proved to be a primary means of protection from viral infections. In the present investigation, growth performance and pathology in breeder vaccinated and non-vaccinated chicks were studied after a challenge with the homologous malabsorption strain of the reovirus. Improvements in growth performance [mean live body weight, feed conversion ratio, broiler performance efficiency index, and protein efficiency index] in breeder vaccinated chicks were compared with non-vaccinated breeder chicks. The non vaccinated chicks showed various signs and lesions indicative of the reoviral malabsorption syndrome [MAS], whereas the vaccinated chicks showed very minimal alterations, demonstrating that the vaccination of breeders with homologous strains of the reovirus is profitable, and can help to increase the performance of broiler birds


Subject(s)
Animals , Reoviridae , Vaccination , Birds , Virus Inactivation , Chickens
9.
Article in English | WPRIM | ID: wpr-296528

ABSTRACT

<p><b>OBJECTIVE</b>This paper aims to investigate the apoptotic effect of inactivated Sendai virus (hemagglutinating virus of Japan-enveloped, HVJ-E) on murine melanoma cells (B16F10) and the possible mechanisms involved in the putative apoptotic reactions.</p><p><b>METHODS</b>B16F10 cells were treated with HVJ-E at various multiplicities of infection (MOI), and the reactive oxygen species (ROS), cell viability, and apoptosis were measured. Next, the roles of ROS in the regulation of Bcl-2/Bax and the activation of mitogen-activated protein kinase (MAPK) pathways in HVJ-E-treated B16F10 cells were analyzed. To further evaluate the cytotoxic effect of HVJ-E-generated ROS on B16F10 cells, HVJ-E was intratumorally injected, both with and without N-acetyl-L-cysteine (NAC), into melanoma tumors on BALB/c mice. Tumor volume was then monitored for 3 weeks, and the tumor proteins were separated for immunoblot assay.</p><p><b>RESULTS</b>Treatment of B16F10 cells with HVJ-E resulted in a dose-dependent inhibition of cell-viability and an induction of apoptosis. The latter effect was associated with the generation of ROS. Inhibition of ROS generation by NAC resulted in a significant reduction of HVJ-E-induced Erk1/2, JNK, and p38 MAPK activation. Additionally, ROS inhibition caused a decrease in the Bcl-2/Bax ratio as well as promoting activation of apoptosis both in vitro and in vivo.</p><p><b>CONCLUSION</b>These results suggest that HVJ-E possesses potential anticancer activity in B16F10 cells through ROS-mediated mitochondrial dysfunction involving the MAPK pathway.</p>


Subject(s)
Animals , Apoptosis , Cell Line, Tumor , Mice , Mitogen-Activated Protein Kinase 1 , Genetics , Metabolism , Reactive Oxygen Species , Metabolism , Respirovirus Infections , Virology , Sendai virus , Physiology , Virus Inactivation
10.
Braz. j. biol ; 75(4,supl.2): 37-42, Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769600

ABSTRACT

The present study analyzed the efficiency of the photo-electro-oxidation process as a method for degradation and inactivation of adenovirus in water. The experimental design employed a solution prepared from sterile water containing 5.107 genomic copies/L (gc/L) of a standard strain of human adenovirus type 5 (HAdV-5) divided into two equal parts, one to serve as control and one treated by photo-electro-oxidation (PEO) for 3 hours and with a 5A current. Samples collected throughout the exposure process were analyzed by real-time polymerase chain reaction (qPCR) for viral genome identification and quantitation. Prior to gene extraction, a parallel DNAse treatment step was carried out to assess the integrity of viral particles. Integrated cell culture (ICC) analyses assessed the viability of infection in a cell culture. The tested process proved effective for viral degradation, with a 7 log10 reduction in viral load after 60 minutes of treatment. The DNAse-treated samples exhibited complete reduction of viral load after a 75 minute exposure to the process, and ICC analyses showed completely non-viable viral particles at 30 minutes of treatment.


Resumo O presente estudo analisou a eficiência do processo de fotoeletrooxidação como metodologia para a degradação e inativação de adenovírus em água. A concepção experimental emprega uma solução preparada a partir de água estéril contendo 5,107 cópias genômicas/L (gc/L) de uma amostra padrão de adenovírus humano tipo 5 (HAdV-5), dividida em duas partes iguais, uma para servir como controle e outra tratada por fotoeletrooxidação (PEO) durante 3 horas e com uma corrente de 5A. As amostras recolhidas durante o processo de exposição foram analisadas por PCR quantitativo em tempo real (qPCR) para identificação e quantificação do genoma viral. Antes da extração de ácidos nucleicos, um passo de tratamento com DNAse paralelo foi realizado para avaliar a integridade das partículas virais. Um ensaio de qPCR integrado à cultura de células (ICC-qPCR) permitiu analisar a viabilidade de infecção em uma cultura de células. O processo mostrou-se eficaz testada para a degradação viral, com uma redução de 7 log10 da carga viral após 60 minutos de tratamento. As amostras tratadas com DNAse exibiram redução completa da carga viral após uma exposição de 75 minutos ao processo, e a análise de ICC-qPCR mostrou partículas virais completamente não-viáveis ​​em 30 minutos de tratamento.


Subject(s)
Adenoviruses, Human/isolation & purification , Virus Inactivation , Waste Disposal, Fluid/methods , Water Purification/methods , Electrochemical Techniques , Oxidation-Reduction , Photolysis , Real-Time Polymerase Chain Reaction
11.
Chinese Journal of Virology ; (6): 500-506, 2015.
Article in Chinese | WPRIM | ID: wpr-296256

ABSTRACT

Enterovirus 71 (EV71) is a major agent of hand, foot and mouth disease that can cause a severe burden of disease to children. To identify an effective method for the control and prevention of EV71, we studied the effect of exposure to heat and ultraviolet (UV) light upon EV71 inactivation. We found that exposure to 50 degrees C could not inactivate the infectivity of EV71. However, exposure to 60 degrees C and 70 degrees C could inactivate EV71 effectively. EV71 could be inactivated after exposure to UV light at a distance between the sample and a lamp of 30 cm for 30 min or 60 min because viral genomic RNA was destroyed. However, fetal bovine serum (FBS) could attenuate the inactivation proffered by heat and UV light. Attenuation effects of FBS were correlated positively with FBS concentration. Hence, EV71 can be inactivated by exposure to heat and UV light, and our results could provide guidance on prevention of the spread of EV71.


Subject(s)
Disinfection , Methods , Enterovirus A, Human , Genetics , Physiology , Radiation Effects , Enterovirus Infections , Virology , Hot Temperature , Humans , Ultraviolet Rays , Virus Inactivation , Radiation Effects
12.
Braz. j. microbiol ; 45(1): 209-214, 2014. graf, tab
Article in English | LILACS | ID: lil-709459

ABSTRACT

The bovine viral diarrhoea virus (BVDV) is suggested as a model for antiviral studies of the hepatitis C virus (HCV). The antiviral activity of the essential oil of Ocimum basilicum and the monoterpenes camphor, thymol and 1,8-cineole against BVDV was investigated. The cytotoxicities of the compounds were measured by the MTT (3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) test, and the antiviral activities were tested by the plaque reduction assay. The oil or compounds were added to the assay in three different time points: a) pre-treatment of the virus (virucidal assay); b) pre-treatment of the cells; or c) post-treatment of the cells (after virus inoculation). The percentage of plaques inhibition for each compound was determined based on the number of plaques in the viral control. The results were expressed by CC50 (50% cytotoxic concentration), IC50 (inhibitory concentration for 50% of plaques) and SI (selectivity index = CC50/IC50). Camphor (CC50 = 4420.12 µgmL-1) and 1,8-cineole (CC50 = 2996.10 µgmL-1) showed the lowest cytotoxicities and the best antiviral activities (camphor SI = 13.88 and 1,8-cineol SI = 9.05) in the virucidal assay. The higher activities achieved by the monoterpenes in the virucidal assay suggest that these compounds act directly on the viral particle.


Subject(s)
Antiviral Agents/pharmacology , Monoterpenes/pharmacology , Ocimum basilicum/chemistry , Oils, Volatile/pharmacology , Pestivirus/drug effects , Plant Extracts/pharmacology , Virus Inactivation , Antiviral Agents/isolation & purification , Antiviral Agents/toxicity , Cell Survival/drug effects , Colorimetry/methods , Microbial Sensitivity Tests , Monoterpenes/isolation & purification , Monoterpenes/toxicity , Oils, Volatile/isolation & purification , Oils, Volatile/toxicity , Pestivirus/growth & development , Plant Extracts/isolation & purification , Plant Extracts/toxicity , Tetrazolium Salts/metabolism , Thiazoles/metabolism , Viral Plaque Assay
13.
Article in English | WPRIM | ID: wpr-343083

ABSTRACT

This paper aimed to study the ability of baicalein to block human cytomegalovirus (HCMV) infection in extravillous cytotrophoblasts (EVT) and its effect on the vasoactive intestinal peptide (VIP) expression in HCMV-infected EVT in vitro. A human trophoblast cell line (HPT-8) was chosen in this study. HCMV with 100 TCID50 was added into culture medium to infect HPT-8 cells, and then HCMV pp65 antigen was assayed by immunofluorescence staining. The infection status was determined by virus titration. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect virus DNA load in the infected cells. The expression of VIP mRNA and protein in the infected cells was measured by qRT-PCR, immunocytochemistry and Western blotting. Concentration of VIP secreted in supernatants was determined by ELISA. Red-stained HCMV pp65 antigens were found in infected HPT-8 cells 48 h after infection. HCMV replicated in large quantity in infected HPT-8 cells 4 days after infection, reaching a peak at day 6 post-infection. After treatment with baicalein, virus DNA load in infected HPT-8 cells was decreased (P<0.05), and the levels of VIP mRNA and protein, and the concentration were raised to the normal (P>0.05). Our study suggested that baicalein exerts a positive effect on the VIP expression in HCMV-infected EVT at maternal-fetal interface.


Subject(s)
Antiviral Agents , Pharmacology , Cell Line , Cytomegalovirus , Physiology , Flavanones , Pharmacology , Humans , Trophoblasts , Cell Biology , Metabolism , Virology , Vasoactive Intestinal Peptide , Metabolism , Virus Inactivation
14.
Rev. bras. hematol. hemoter ; 34(3): 231-235, 2012. ilus, tab
Article in English | LILACS | ID: lil-640874

ABSTRACT

The aim of this study was to conduct a systematic review on the photoinactivators used in hemotherapy, with action on viral genomes. The SciELO, Science Direct, PubMed and Lilacs databases were searched for articles. The inclusion criterion was that these should be articles on inactivators with action on genetic material that had been published between 2000 and 2010. The key words used in identifying such articles were "hemovigilance", "viral inactivation", "photodynamics", "chemoprevention" and "transfusion safety". Twenty-four articles on viral photoinactivation were found with the main photoinactivators covered being: methylene blue, amotosalen HCl, S-303 frangible anchor linker effector (FRALE), riboflavin and inactin. The results showed that methylene blue has currently been studied least, because it diminishes coagulation factors and fibrinogen. Riboflavin has been studied most because it is a photoinactivator of endogenous origin and has few collateral effects. Amotosalen HCl is effective for platelets and is also used on plasma, but may cause changes both to plasma and to platelets, although these are not significant for hemostasis. S-303 FRALE may lead to neoantigens in erythrocytes and is less indicated for red-cell treatment; in such cases, PEN 110 is recommended. Thus, none of the methods for pathogen reduction is effective for all classes of agents and for all blood components, but despite the high cost, these photoinactivators may diminish the risk of blood-transmitted diseases.


Subject(s)
Nucleic Acids , Chemoprevention , Virus Inactivation , Blood Safety
15.
Article in English | WPRIM | ID: wpr-235509

ABSTRACT

<p><b>OBJECTIVE</b>This paper aims to investigate the anti-tumor mechanism of inactivated Sendai virus (Hemagglutinating virus of Japan envelope, HVJ-E) for murine melanoma (B16F10).</p><p><b>METHODS</b>The murine dendritic cells (DCs) were treated with HVJ-E, and then the cytokines secreted from DCs and costimulation-related molecules on DCs were measured. Meanwhile, the expression of β-catenin in HVJ-E treated murine melanoma cells was detected. In addition, HVJ-E was intratumorally injected into the melanoma on C57BL/6 mice, and the immune cells, CTL response and tumor volume were analyzed.</p><p><b>RESULTS</b>HVJ-E injected into B16F10 melanoma obviously inhibited the growth of the tumor and prolonged the survival time of the tumor-bearing mice. Profiles of cytokines secreted by dendritic cells (DCs) after HVJ-E stimulation showed that the number of cytokines released was significantly higher than that elicited by PBS (1P<0.05). The co-stimulation-related molecules on DCs were comparable to those stimulated by LPS. Immunohistochemical examinations demonstrated the repression of β-catenin in B16F10 melanoma cells after HVJ-E treatment. Meanwhile, real-time reverse transcription PCR revealed that HVJ-E induced a remarkable infiltration of CD11c positive cells, chemokine ligand 10 (CXCL10) molecules, interleukin-2 (IL-2) molecule, CD4(+) and CD8(+) T cells into HVJ-E injected tumors. Furthermore, the mRNA expression level of β-catenin in the HVJ-E injected tumors was also down-regulated. In addition, B16F10-specific CTLs were induced significantly after HVJ-E was injected into the tumor-bearing mice.</p><p><b>CONCLUSION</b>This is the first report to show the effective inhibition of melanoma tumors by HVJ-E alone and the mechanism through which it induces antitumor immune responses and regulates important signal pathways for melanoma invasion. Therefore, HVJ-E shows its prospect as a novel therapeutic for melanoma therapy.</p>


Subject(s)
Animals , Cell Line, Tumor , Cytokines , Genetics , Metabolism , Dendritic Cells , Allergy and Immunology , Physiology , Virology , Down-Regulation , Gene Expression Regulation, Neoplastic , Melanoma , Allergy and Immunology , Pathology , Virology , Mice , Mice, Inbred C57BL , Neoplasms, Experimental , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Sendai virus , Physiology , Virus Inactivation , Virus Replication , beta Catenin , Genetics , Metabolism
16.
Article in English | WPRIM | ID: wpr-233161

ABSTRACT

This study examined the anti-hepatitis B virus (HBV) effect of wild-type (WT) vacuolar protein sorting 4B (VPS4B) and its dominant negative (DN) mutant VPS4B-K180Q in vivo in order to further explore the relationship between HBV and the host cellular factor VPS4. VPS4B gene was amplified from Huh7 cells by RT-PCR and cloned into the eukaryotic expression vector pXF3H. Then, the VPS4B plasmid and the VPS4B-K180Q mutation plasmid were constructed by using the overlap extension PCR site-directed mutagenesis technique. VPS4B and HBV vectors were co-delivered into mice by the hydrodynamic tail-vein injection to establish HBV vector-based models. Quantities of HBsAg and HBeAg in the mouse sera were determined by ElectroChemiLuminescence (ECL). HBV DNA in sera was measured by real-time quantitative PCR. Southern blot analysis was used to assay the intracellular HBV nuclear capsid-related DNA, real-time quantitative PCR to detect the HBV-related mRNA and immunohistochemical staining to observe the HBcAg expression in the mouse liver tissues. Our results showed that VPS4B and its mutant VPS4B-K180Q could decrease the levels of serum HBsAg, HBeAg and HBV-DNA. In addition, the HBV DNA replication and the mRNA level of HBV in the liver tissues of treated mice could be suppressed by VPS4B and VPS4B-K180Q. It was also found that VPS4B and VPS4B-K180Q had an ability to inhibit core antigen expression in the infected mouse liver. Furthermore, the anti-HBV effect of mutant VPS4B-K180Q was more potent than that of wild-type VPS4B. Taken together, it was concluded that VPS4B and its DN mutant VPS4B-K180Q have anti-HBV effect in vivo, which helps develop molecular therapeutic strategies for HBV infection.


Subject(s)
ATPases Associated with Diverse Cellular Activities , Adenosine Triphosphatases , Physiology , Animals , Endosomal Sorting Complexes Required for Transport , Physiology , Female , Genes, Dominant , Genetics , Hepatitis B , Metabolism , Virology , Hepatitis B virus , Physiology , Liver , Virology , Mice , Mice, Inbred BALB C , Mutation , Genetics , Virus Inactivation
17.
Rev. cuba. hematol. inmunol. hemoter ; 27(3): 333-341, jul.-set. 2011.
Article in Spanish | LILACS | ID: lil-615362

ABSTRACT

La validación de la capacidad de aclaramiento viral de los procesos de fabricación de productos biológicos constituye un requisito regulatorio en Cuba. Se recomienda introducir la pasteurización en los procesos de producción de la albúmina como un método capaz de inactivar virus; por ello, el objetivo del estudio fue validar la capacidad de inactivación viral de la etapa de pasteurización del proceso de producción de la albúmina humana al 20 y 25 por ciento. Los modelos virales que abarcan los posibles contaminantes de la materia prima, se diluyeron 1:10 en la albúmina en sus 2 concentraciones y se sometieron a tratamiento térmico a 60 °C durante 10 h. Se tomaron muestras a diferentes intervalos de tiempo para la confección de las curvas de cinética de inactivación. Se determinó el factor de reducción aportado por la pasteurización para cada virus. El tratamiento a 60 °C de la albúmina al 20 y 25 por ciento disminuyó significativamente la carga viral inicial con que se retó la etapa, con valores de p< 0,002 y p< 0,021, respectivamente, y se obtuvieron factores de reducción superiores a 4 log del título de todos los virus. La etapa de pasteurización le aportó a la albúmina humana al 20 y 25 por ciento un adecuado nivel de seguridad


The validation of the capacity of viral clearance in the manufacturing processes of biopharmaceuticals is a regulatory requirement in Cuba. It is recommended to introduce the pasteurization in the manufacturing processes of serum albumin as a method of inactivating viruses. The objective of this study was to validate the capacity of viral inactivation in the phase of pasteurization of the 20 percent and 25 percent human albumin production process The viral models covering the possible pollutants of the raw materials were diluted at 1:10 in albumin in 2 concentrations and they were heat-treated at 60 °C for 10 h. Several samples at different time intervals were taken to design the inactivation kinetic curves. The reduction factor of pasteurization for each virus was estimated. The treatment of 20 percent and 25 percent albumin at 60 °C decreased significantly the initial viral load in the stage, with p< 0.002 and p< 0.021 respectively. The reduction factors exceeded 4 log of the titers of all viruses. The stage of pasteurization gave adequate level of safety to the 20 percent and 25 percent human albumin


Subject(s)
Humans , Serum Albumin/analysis , Serum Albumin/biosynthesis , Pasteurization/methods , Virus Inactivation/ethics , Validation Studies as Topic
18.
Chinese Journal of Biotechnology ; (12): 1541-1548, 2011.
Article in Chinese | WPRIM | ID: wpr-304547

ABSTRACT

Four out of 10 patients of X-linked severe combined immunodeficiency (X-SCID) were finally developed leukemia after receiving the treatment of gene therapy delivered by gamma-retroviral vectors. This is due to the vector integrated to the proximity of lmo2 etc proto-oncogene promoters, leading to the activation of onco-gene expression, which raises the concern of the bio-safety of gene therapy vectors. Lentiviral vectors, especially self-inactivating lentiviral vectors, are considered to be much safer than gamma-retroviral vectors. However self-inactivating lentiviral vectors also have encountered with some unsafe factors and one of them is the problem of transcriptional "read-through" . During the past years, achievements have been made to reduce lentiviral vector transcriptional read-through, which are reviewed herein.


Subject(s)
Animals , Genetic Therapy , Methods , Genetic Vectors , Genetics , Humans , Lentivirus , Genetics , Metabolism , Promoter Regions, Genetic , Proto-Oncogene Proteins , Genetics , Transcription, Genetic , Genetics , Virus Inactivation , Virus Integration
19.
Article in Chinese | WPRIM | ID: wpr-244949

ABSTRACT

This study was aimed to evaluate the efficacy of riboflavin photochemical inactivation of virus in red blood cells by using animal models. human cytomegalovirus (HCMV) plus red blood cells were used as indicator, 30 BALA/c mice were divided into the experimental group (n = 10), virus control group (n = 10), visible light control group (n = 5) and red blood cell control group (n = 5). Mice in experimental group were inoculated with red blood cells inactive by the riboflavin photochemical, mice in virus control group was injected with red blood cells without riboflavin photochemical inactivation treatment, and mice in light control group was infused with red blood cells irradiated by visible light, and mice in red blood cells control group was injected with normal red blood cells. The virus was isolated in vitro from mice of various groups, the HCMV UL83 gene was detected by PCR, the PP65 antigen was identified by indirect immunofluorescence. The results indicated that the virus isolation, PCR detection and indirect immunofluorescence identification all showed positive in virus control group and visible light control group, while the results of detection in experimental and red blood cell control groups were negative. It is concluded that riboflavin photochemical viral inactivation of red blood cells is effective.


Subject(s)
Animals , Erythrocytes , Virology , Humans , Mice , Mice, Inbred BALB C , Models, Animal , Photochemistry , Riboflavin , Pharmacology , Virus Inactivation
20.
Braz. j. pharm. sci ; 46(4): 777-783, Oct.-Dec. 2010. ilus, graf, tab
Article in English | LILACS | ID: lil-622878

ABSTRACT

Highly purified intravenous immunoglobulin G concentrate (IV IgG) was produced with the use of polyethylene glycol associated to a single-stage precipitation by ethanol, instead of the classic Cohn-Oncley process, which employs cold alcohol as the precipitating agent, in a three-stage process. Precipitation of crude fraction containing more than 95% of immunoglobulin G was performed by liquid chromatography with a cation exchanger, CM-Sepharose, as a stationary phase. During the process, the product was subjected to two-stage viral inactivation. The first stage was performed by the action of sodium caprylate, 30 mM at pH 5.1+/- 0.1, and the second stage was performed by the action of a solvent-detergent mixture. The finished product was formulated at 5% with 10% sucralose as the stabilizing agent. The process yields 3.3g of IgG/liter of plasma. The finished product analysis showed an anti-complementary activity lower than 1CH50. Polymer and aggregate percent levels were lower than 3% in the five batches studied. The analysis of neutralizing capacity showed the presence of antibacterial and antiviral antibodies in at least three times higher concentrations than the levels found in source plasma. The finished product fulfilled all purity requirements stated in the 4th edition of the European pharmacopeia.


Obteve-se concentrado de imunoglobulina G intravenosa IgGIV, altamente purificado, utilizando-se polietilenoglicol associado a uma única etapa de precipitação por etanol, em substituição ao tradicional método descrito por Cohn-Oncley, que emprega, em três etapas, o mesmo álcool resfriado, como agente precipitante. A purificação da fração bruta contendo mais de 95% de imunoglobulina G foi realizada utilizando-se cromatografia líquida com um trocador de cátion, a CM-Sepharose, como fase estacionária. Durante o processamento o produto foi submetido a dupla inativação viral sendo a primeira pela ação do caprilato de sódio, 30 mM a pH 5,1+/- 0,1 e a segunda por ação de mistura de solvente/detergente. O produto acabado foi formulado a 5% utilizando-se sucralose 10% como estabilizante. O rendimento da metodologia foi de 3,3g de IgG/litro de plasma. A análise do produto acabado demonstrou atividade anti-complementar inferior a 1CH50. O valor percentual de polímeros e agregados em cinco lotes realizados foi inferior a 3%. O estudo da capacidade de neutralização demonstrou a presença de anticorpos anti-bacterianos e anti-virais em concentração pelo menos três vezes maior que o plasma de origem. O produto acabado apresentou conformidade com todos os requisitos de pureza dispostos na farmacopéia européia IV edição.


Subject(s)
Immunoglobulins, Intravenous/isolation & purification , Solutions/analysis , Virus Inactivation , Chromatography, Ion Exchange , Good Manipulation Practices , Polyethylene/blood , Ultrafiltration/methods
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