Cytotoxic effect of the immunotoxin constructed of the ribosome-inactivating protein curcin and the monoclonal antibody against Her2 receptor on tumor cells.

The toxicity of the curcin on cancer cells allows to consider this protein as the toxic component of an immunotoxin directed to Her2, which is associated with cancer. Reductive amination was proposed to conjugate curcin and an anti-Her2; the binding was tested using Polyacrylamide gel electrophoresis, western blot, and immunocytochemistry. The in vitro cytotoxicity of curcin and the immunotoxin was assessed on breast cancer cell lines SK-BR-3 (Her2(+)) and MDA-MB-231 (Her2(-)). IC50 values for curcin were 15.5 ± 8.3 and 18.6 ± 2.4 µg/mL, respectively, statistically equivalent (p < 0.05). While to the immunotoxin was 2.2 ± 0.08 for SK-BR-3 and 147.6 ± 2.5 µg/mL for MDA-MB-231. These values showed that the immunotoxin was seven times more toxic to the SK-BR-3 than curcin and eight times less toxic to the MDA-MB-231. The immunotoxin composed of curcin and an antibody against Her2 and constructed by reductive amination could be a therapeutic candidate against Her2(+) cancer.

Anti-hyperglycemic effect, inhibition of inflammatory cytokines expression, and histopathology profile in streptozotocin-induced diabetic rats treated with Arracacia tolucensis aerial-parts extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Arracacia tolucensis is a medicinal plant used in northeast of Mexico as a remedy to treat people with Diabetes mellitus (DM); however, there are no scientific studies that support this information. Thus, we evaluated the anti-hyperglycemic effect of the hexane, ethyl acetate and ethanol extracts from aerial parts in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: DM was induced in Wistar male rats by single intraperitoneal injection of streptozotocin (STZ 50mg/kg). After STZ-induction, hyperglycemic rats were treated with all three extracts orally at a single dose (250 mg/kg) each 48 h for 21 days. Glibenclamide (1mg/kg) was used as a reference drug. The fasting blood glucose levels, the hematic biometry and biochemical profiles, and the inhibition of inflammatory cytokines expression were estimated. Histopathology analysis of pancreas, liver, spleen, and kidney tissue was carried out. RESULTS: Ours results showed that ethyl acetate extract decreased blood glucose levels significantly (75%, p< 0.05) when compared to diabetic rats and controlled the body weight loss; the lipids level did not change, but the enzyme levels of aspartate aminotransferase and alanine aminotransferase decreased significantly (60.83% and 66.16%, respectively, p< 0.05) and inhibited the expression of inflammatory cytokines,with respect to diabetic rats. Histopathology injury was not observed; by contrast repair of islet of Langerhans was exhibited. CONCLUSION: These results validate the use of Arracacia tolucensis as a treatment against DM and suggests it is suitable to continue studies for its safe therapeutic use.

Immunological evaluation of captive green sea turtle (Chelonia mydas) with ulcerative dermatitis.

Ulcerative dermatitis (UD) is common in captive sea turtles and manifests as skin erosions and ulcers associated with gram-negative bacteria. This study compared clinically healthy and UD-affected captive turtles by evaluating hematology, histopathology, immunoglobulin levels, and delayed-type hypersensitivity assay. Turtles with UD had significantly lower weight, reduced delayed-type hypersensitivity (DTH) responses, and higher heterophil:lymphocyte ratios. This study is the first to assay DTH in green turtles (Chelonia mydas) and suggests that UD is associated with immunosuppression.

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas.

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6k Da band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demonstration of CD3+ cells in reptiles using specific antibodies.

Caracterización antigénica de cepas del virus del ectima contagioso (orf), sus interacciones y sus relaciones con parapoxvirus bovinos de México / Antigenic characterization of contagious ecthyma virus (orf) in Mexico, their relationships and interactions with bovine parapoxvirus samples

Treinta y tres muestras de costra positivas a ectima contagioso (orf) (19 de caprino y 14 ovinas) y dos de parapoxvirus bovinos (una de estomatitis papular y otra de seudoviruela) fueron analizadas antigénicamente y comparadas por doble inmunodifusión (DD) y contrainmunoelectroforesis (CIE). Las muestras bovinas fueron obtenidas de animales que convivían con rebaños de borregos y cabras. La presencia de partículas virales fue confirmada por microscopía electrónica en todos los casos. Las pruebas fueron corridas usando un suero policlonal antiectima preparado en conejo. En la prueba de DD la muestra homóloga produjo tres líneas de precipitación, las otras muestras variaron entre dos y una líneas de precipitación y algunas muestras no formaron ninguna, se observaron diferentes resultados con las pruebas de DD y CIE. Las cepas bovinas formaron dos líneas de precipitación, una de identidad entre ellas y otra en identidad con la muestra homóloga de orf caprino y con una de orf ovino, por DD. La composición de 15 muestras de orf (6 ovinas y 9 caprinas) y dos bovinas fueron examinadas por electroforesis, y 7 de ellas evaluadas antigénicamente por inmunotransferencia, usando el suero policlonal de conejo. La muestra homóloga fue más compleja, pues mostró 12 proteínas. La mayor variación en los corrimientos electroforéticos de las muestras se observó entre las proteínas de los pesos de 37 kd a 44 kd. Una proteína de 55 kd se identificó en todas las muestras, mientras que una de 9.5 kd se evidenció en 13 muestras y otra de 17 kd en 12, incluyendo las bovinas. Por inmunotransferencia fueron reconocidas dos proteínas comunes en las siete muestras analizadas con pesos de 55 y 54 kd, otras dos proteínas de 45 y 35 kd fueron reconocidas por el suero de conejo en cinco de las muestras. Estas proteínas pueden explicar las reacciones cruzadas observadas entre las muestras, en las pruebas de DD y CIE y ser de importancia en el reconocimiento inmune de estos virus.

Patrones electroforéticos y antigénicos de los serotipos 1, 2, 5 y 7 de Actinobacillus pleuropneumoniae / Elecrtophoretic and antigenic patterns of Actinobaccillus pleuropneumoniae serotypes 1, 2, 5 and 7

Se estudiaron extractos bacterianos solubilizados con Triton X-100 de los serotipos 1,2, 5 y 7 de Actinobacillus pleuropneumoniae. Los extractos se analizaron mediante electroforesis en geles de sodio-duodecil-sulfato (SDS) poliacrilamida e inmunotransferencia con sueros hiperinmunes de conejo y 9 sueros de cerdo A. pleuropneumoniae positivos. Los patrones electroforéticos mostraron las siguientes proteínas comunes a todos los serotipos: 26-29, 30-33, 43, 50, 66 y 81-83 Kd. En cuanto a los patrones antigénicos con sueros hiperinmunes de conejo, se observó un antígeno de 30-31 Kd de peso molecular reconocido por todos los antisueros con todos los serotipos (con excepción del serotipo 7, cuando reaccionó con el antisuero contra el serotipo 5).En relación con los antígenos reconocidos por sueros de cerdo, se onservó un grupo de antígenos cuyo peso molecular osciló entre 29 y 33 Kd, reconocido por todo los sueros con todos los serotipos, salvo la reacción de 3 sueros con el serotipo 2.