ABSTRACT
<p><b>OBJECTIVE</b>To find a suitable cell line for hepatitis A antigen expressed by vaccinia virus vector and to find a way of inactivation and preservation of the HAV recombinant antigen. Methods Series of cell lines such as K4,143, HEL, Hep-2 and Vero were inoculated with vaccinia virus that can express HAV recombinant antigen. ELISA was used to determine the contents of expression antigen. The characterization of the HAV antigen expressed by vaccinia virus was then analyzed after it was treated with different methods.</p><p><b>RESULTS</b>The expression of HAV recombinant antigen in K4,143 and HEL cell lines was a little more than expression in Hep-2 and Vero cell lines. The antigenicity is obviously higher when HAV recombinant antigen was inactivated by beta-propiolactone other than it was inactivated by formalin. It was best to preserve the prepared HAV recombinant antigen under -40 degrees C condition.</p><p><b>CONCLUSIONS</b>The application of vaccinia virus vector in hepatitis A antigen preparation was very useful and promising.</p>
Subject(s)
Animals , Humans , Cell Line , Enzyme-Linked Immunosorbent Assay , Formaldehyde , Pharmacology , Genetic Vectors , Hepatitis A Antigens , Genetics , Allergy and Immunology , Hepatitis A Vaccines , Allergy and Immunology , Propiolactone , Pharmacology , Recombinant Proteins , Allergy and Immunology , Vaccines, Synthetic , Allergy and Immunology , Vaccinia virus , GeneticsABSTRACT
The inhibition of serum alanine and aspartate aminotransferases [SALT and SAST] by beta nitropropionic acid [beta NPA], toxic metabolite of some fungi higher plants] in vitro was studied. The results indicated that both SALT and SAST were competitively inhibited by beta NPA and the enzymes recovered their original activity by dialysis, indicating that the inhibitory effect of beta NPA is reversible. The inhibition of both SALT and SAST by beta NPA was found to be slow and showed the characteristic of a first order reaction up to 30 minutes. The rate constants characterizing this inhibition, namely: the binding constant [k[B]] [90 uM and 225 uM for SALT and SAST, respectively] and bimolecular velocity of inhibition ki [666 and 714 [M min][-1] for SALT and SAST, respectively were determined. Kn [rate of nitrification of the enzymes] for SALT and SAST were 0.06/min and 0.18/min, respectively
Subject(s)
Aspartate Aminotransferases , Propionates , Enzyme Inhibitors , PropiolactoneABSTRACT
A Cross-sectional study was carried out to determine the profile of animal bite cases reporting to the Anti Rabies clinic of Sassoon Hospital, rune. The data was collected using pretested questionnaire. All 250 cases who reported during the period of study were included in the analysis. The male female ratio was 1.98 : 1. Children in the age group 0-14 years were the victims in 132 (52.8%) cases. Dog was the biting animal in 94.4% cases, followed by cat (2.4%), Jackal (1.2%), mongoose (1.2%), monkey (0.4%) and horse (0.4%). Of the 236 dog bite cases 30% of bites were inflicted by pet dogs of which only 38.02% were immunized. The wound was washed with soap and water in only 3.6% of cases. 64.8% of the bites were on the lower extremity and 63.2% of cases reported within 24 hours of the bite. Of the 247 cases administered Beta Propio Lactone (BPL) inactivated vaccine only 18.8% did not have any local reaction and 58.3% had one or more systemic reaction. A three pronged strategy has been recommended to reduce the burden of morbidity and mortality associated with rabies.
Subject(s)
Adolescent , Adult , Aged , Animals , Bites and Stings/epidemiology , Child , Child, Preschool , Cross-Sectional Studies , Disinfectants/administration & dosage , Dogs , Female , Humans , India/epidemiology , Infant , Male , Middle Aged , Propiolactone/administration & dosage , Surveys and Questionnaires , Rabies/prevention & control , Rabies Vaccines , Sex DistributionABSTRACT
<p><b>OBJECTIVE</b>To study the method for cultivation and inactivation of SARS-CoV.</p><p><b>METHODS</b>In order to choose the sensitive cell strain and the best infection dose of the virus, Vero, Vero-E6 and 2BS cell lines were infected with SARS-CoV. The cultivation temperature was selected among 25 degrees C, 33 degrees C and 37 degrees C. The best inactivating time and effect were observed with beta-propiolactone whose concentration ranged from 1:2000 to 1:20,000 at room temperature.</p><p><b>RESULTS</b>Vero and Vero-E6 cell lines were sensitive to SARS-CoV. The cytopathic changes of the cells were 75% at 37 degrees C in 5 percent CO2 incubator after infection. SARS-CoV was inactivated completely in beta-propiolactone (1:4000). The toxicity of beta-propiolactone was hydrolyzed completely when the inactivated virus was cultured for 16 hours at 2 degrees C, 8 degrees C and in water bath for 2 hours at 37 degrees C.</p><p><b>CONCLUSION</b>The titer of SARS-CoV was the highest when it was cultured in Vero or Vero-E6 cells for 72 hours at 37 degrees C in 5 percent CO2 incubator. SARS-CoV was inactivated completely in beta-propiolactone when its concentration was 1:4000 and the interaction time was 1 hour.</p>
Subject(s)
Animals , Chlorocebus aethiops , Dose-Response Relationship, Drug , Propiolactone , Pharmacology , Severe acute respiratory syndrome-related coronavirus , Temperature , Time Factors , Vero Cells , Virus InactivationABSTRACT
To investigate the relationship between the morphologic changes and the expression of keratin and proto-oncogene induced by Beta-propiolactone (BPL), we assessed on the expression of keratins (K8, K10, K13) and proto-oncogenes (c-fos, c-jun, c-myc) in human HaCaT cell line. The cells were treated with 0, 0.1, 1 microgram/ml BPL for 2 or 6 hours. Morphologic studies revealed that BPL changed the cells into fibrocyte-shaped, caused highly lobulated nuclei and reduced desmosomes in their number. Findings of immunofluorescence and Northern blotting indicated that BPL consistently decrease expression of K10 representing a normal differentiation marker of keratinocytes, while increasing expression of K8 and K13 associated with a pathologic differentiation. This reagent also up-regulated expression of c-fos and c-jun, and down-regulated expression of c-myc. Together with staining for each keratin or proto-oncogene and DNA content in flow cytometry, BPL increased K8 expression dramatically at S-G2-M phase. The induction of c-Fos at S-G2-M phase appeared within 2 hours, and c-Jun gradually occurred. However, c-Myc was inhibited regardless of phases of cell cycle. In conclusion, these changes caused by BPL demonstrate a close relationship between the morphologic evidence and the altered expression of each keratin and proto-oncogene.
Subject(s)
Humans , Blotting, Northern , Cell Cycle , Cell Line , Desmosomes , DNA , Flow Cytometry , Fluorescent Antibody Technique , Keratinocytes , Propiolactone , Proto-OncogenesABSTRACT
A reducao da ativacao do complemento atraves de uma alteracao do fragmento Fc das imunoglobulinas pela beta-propiolactona foi obtida em soros hiperimunes equinos antivirus rabico, venenos Bothrops e toxina difterica. Os resultados foram avaliados por teste de anafilaxia em cobaias, e comparados com aqueles obtidos com os mesmos soros purificados por precipitacao salina (sulfato de amonio), seguidos ou nao por digestao enzimatica com pepsina. Os niveis de pureza proteica foram para o soro antibotropico de 184,5 mg/g e 488,5 mg/g tratado pela beta-propiolactona e digeridos pela pepsina, respectivamente...(au)
Subject(s)
Humans , Animals , Complement Activation/drug effects , Immune Sera/isolation & purification , Propiolactone/pharmacology , Complement Fixation Tests , Complement Pathway, Alternative , Diphtheria Toxin , Immunization , Immunoglobulin Fragments/analysis , Rabies Vaccines , Snake VenomsABSTRACT
Cytogenetic effects of the two inactivated viral vaccines (polio and antirabies) were studied in adult male mice by the micronucleus test. Polio salk vaccine did not induce micronuclei formation at both human (0.5 ml) and 1/5th human doses. Antirabies vaccine induced micronuclei in poly and total erythrocytes only at human dose of 2 ml. Beta-propiolactone (BPL) induced micronuclei at higher dose of 5.7 mg, but not at 0.57 mg (approximate concentration present in 2 ml of rabies vaccine). The P/N ratio was not affected in vaccinated and BPL inoculated animals. Antirabies vaccine induced micronuclei percentage was more than the BPL value.
Subject(s)
Animals , Bone Marrow/drug effects , Bone Marrow Cells , Erythrocytes/ultrastructure , Male , Mice , Micronucleus Tests , Poliovirus Vaccine, Inactivated/pharmacology , Propiolactone/pharmacology , Rabies Vaccines/pharmacology , Vaccines, Inactivated/pharmacologyABSTRACT
Foram analisados os efeitos da beta-propiolactona em testes hematológicos. A incubaçäo do sangue com este agente resultou numa significante diminuiçäo da VHS e num aumento da concentraçäo da hemoglobina globular média e da contagem dos bastöes. A realizaçäo dos esfregaços logo após a adiçäo da droga evita este artefato. Os demais parâmetros permaneceram inalterados
Subject(s)
Humans , Antiviral Agents , HIV/drug effects , Laboratory Infection/prevention & control , Propiolactone/pharmacologyABSTRACT
Foi estudada a interferência da beta-propiolactona (BPL) em testes imunológicos in vitro. As dosagens das imunoglobulinas, dos componentes C3 e C4 do complemento e a a pesquisa de fatores antinucleares näo foram afetadas pela presença deste agente químico. No entanto, foram observadas alteraçöes significativas nas determinaçöes do título do complemento hemolítico total (CH50), do percentual de linfócitos totais e de suas subpopulaçöes, e na pesquisa de células LE
Subject(s)
Humans , HIV/drug effects , Immunoglobulins/analysis , In Vitro Techniques , Laboratory Infection/prevention & control , Propiolactone/pharmacologyABSTRACT
Foram estudados os efeitos da beta-propiolactona em testes bioquímicos, não sendo observadas alterações significantes em qualquer dos testes realizados.
Subject(s)
Humans , Blood Chemical Analysis , HIV/drug effects , Propiolactone/pharmacology , HIV Infections/prevention & controlABSTRACT
Foram estudados os efeitos resultantes da adição da beta-propiolactona (BPL) ao sangue total e soro nas dosagens de íons, na capacidade de fixação do ferro (TIBC), nas mucoproteínas, na eletroforese de proteínas e em algumas provas de funções hepáticas e pancreáticas. A adição desta substância ao sangue pode levar à hemólise, com conseqüentes alterações de análises bioquímicas. De todos os testes estudados apenas as dosagens de mucoproteínas nas alíquotas de soro contendo BPL apresentaram alterações com significado estatístico. Os autores recomendam o uso da BPL antes da realização das análises bioquímicas de rotina em soros de indivíduos infectados pelo HIV ou pertencentes a grupos de risco.
Subject(s)
Humans , Blood Chemical Analysis , HIV , Propiolactone/pharmacology , Blood Protein Electrophoresis , Liver/physiology , HIV Infections/prevention & control , Ions , Iron/metabolism , Mucoproteins , Pancreas/physiologyABSTRACT
Neste trabalho säo revistos os principais métodos físicos e químicos empregados para a inativaçäo do HIV. Discutem-se as características físico-químicas e a toxicidade da beta-propiolactona para, ao final, sugerir-se que as amostras de sangue, soro e plasma destinadas a exames laboratoriasi sejam previamente tratadas por esta substância