Obtención y evaluación del líquido ascítico murino hemoclasificador anti-B en las líneas de ratones Ofbalb (F1) y Balb/c / Obtainment and evaluation of the murine ascitic liquid anti-B hemo-classificatory in Ofbalb (F1) and Balb/c mice strains

La línea de ratón Balb/c ha sido utilizada tradicionalmente para la producción de líquido ascítico murino; sin embargo, actualmente se están introduciendo líneas híbridas, como la Ofbalb (F1).En el presente trabajo, se evaluó el comportamiento del hibridoma C6G4 (anti-B) expandido en ratones de la línea Balb/c y en la línea híbrida Ofbalb (F1). De cada línea se conformaron cuatro grupos, a los que se les administró por vía intraperitoneal las concentraciones celulares del inóculo: 1; 2,5; 3 y 4 x 10(6) células/0,5 mL/ratón. Los parámetros analizados fueron, el por ciento de prendimiento, el volumen y el rendimiento del líquido ascítico murino por ratón. Se demostró que con la línea Balb/c se obtienen mejores resultados para este hibridoma, en particular. El inóculo más adecuado fue el correspondiente a 2,5 x 10(6) células. La línea híbrida Ofbalb (F1) con tratamientos similares no mostró diferencias estadísticamente significativas en los parámetros considerados

The Balb/c mouse strain has been traditionally used for the production of murine ascitic fluid; however, nowadays hybrid strains are introduced including the Ofbalb (F1). The aim of present paper was to assess the behavior of C6G4 (anti-B) hybridoma scattered in mice of Balb/c strain and in the Ofbalb (F1) hybrid strain. From each strain four groups were created which received by intraperitoneal route the cell concentrations of the inoculum: 1; 2,5; 3 and 4 x 10(6) cells/0,5 mL/mouse. The analyzed parameters were the seizure percentage, the volume and the performance of murine ascitic fluid by mouse. It was demonstrated that with the Balb/c strain better results are obtained for this hybridoma in particular. The more appropriate inoculum was that corresponding to 2,5 x 106 cells. The Ofbalb (F1) hybrid strain with similar treatments not showed statistically significant differences in the analyzed parameters

Pérdida de donaciones sanguíneas por positividad de virus de inmunodeficiencia humana en la provincia de Santiago de Cuba 2001- 2005 / Loss of blood donations due to HIV positive testing in Santiago de Cuba province 2001 - 2005

Se efectuó un estudio descriptivo y retrospectivo de las 197 723 donaciones de sangre efectuadas entre 2001 y 2005 en la provincia de Santiago de Cuba, con vista a mejorar la calidad del tamizaje de ese líquido y determinar la frecuencia del síndrome de inmunodeficiencia adquirida en la población donante, procedente de los diferentes centros de extracciones ubicados en los diferentes municipios y recogidas en el Banco de Sangre Provincial Renato Guitart Rosell. Se detectó la presencia del virus en 1 865 casos (0,95 por ciento), equivalente a 932 litros de sangre perdidos.

A descriptive and retrospective study of the 197 723 blood donations between 2001 and 2005 in Santiago de Cuba province was made, with the aim of improving the quality of the screening of that liquid and of determining the frequency of the acquired immunodeficiency syndrome in the donating population, from the different donation institutions located in the different municipalities and from collections in Renato Guitart Rosell Provincial Blood Bank. The presence of the virus was detected in 1 865 cases (0,95 per cent), equivalent to 932 liters of blood lost.

Non-enzymatic in vitro DNA labeling and label immunoquantification.

In the present work, a label immunoquantification procedure was developed in order to determine the number of markers introduced into DNA. A non-enzymatic, in vitro labeling method for introducing the p-bromobenzoyl radical (label), through transamination and acylation reactions of the cytidine nucleotides in calf thymus DNA, was carried out. Three spacer arms with different lengths were used for separating the label from the nucleotide and three labeled DNA were obtained. Anti-p-bromobenzoyl chicken IgY polyclonal antibodies were obtained. The antibodies detected the label, into three-labeled DNA, with different sensitivities, in relation to spacer arm length used. About 3-11 labels per 4 x 10(6) bases into thermally denatured DNA were immunoquantified.

Extraction of a monospecific Coombs-reagent from chicken eggs.

During the last ten years the extraction of specific antibodies (ab) from the yolk of eggs of immunised chickens is more and more accepted as an useful alternative to the immunisation of mammals. The subject of this work is the immunisation of chickens with human IgG and the extraction of specific anti human IgG ab from egg yolk in order to obtain monospecific Coombs reagent. 12 Leghorn hens (25 weeks old) were immunised with intact human IgG (INTACGLOBIN). The chickens were immunised with 100 microg IgG/animal once per week for a period of seven weeks. The highest titre was observed after the 5th immunisation, the following immunisations achieved no further titre increase. The IgY purification was performed according to the method of Akita and Nakai (1993). The resulting IgY preparation was tested for the presence of hetero-agglutinine by means of direct agglutination using human erythrocytes of all blood groups. Thereafter 58 blood donors were tested by means of direct or indirect Coombs-test using a reference reagent (DAKO) and a Coombs reagent isolated from chicken eggs (IgY antibodies). No differences have been found between the results obtained using both Coombs reagents. Presented results show that there is a possibility to produce Coombs-reagent in chickens. Advantages of this method are: 1) non invasive antibody sampling by egg collection instead of bleeding the animal (refinement of antibody production); 2) decreasing amount of animals necessary to produce high amounts of reagent; 3) IgY-preparation contains no hetero-agglutinine in contrast to serum ab from mammals, therefore additional step in reagent production e.g. the absorption of hetero-agglutinins is not necessary.