Facilitating the gut brain axis by probiotic bacteria to modulate neuroimmune response on lead exposed zebra fish models

Unravelling the efficacy of gut biome has a major impact on health. An unbalanced microbiome composition is linked to many common illnesses such as gut dysbiosis, mental deformities and immunological imbalance. An optimistic influence on the gut biome can be made by consumingprobiotics. This would stimulate neuroprotection and immunomodulation intended by heavy metals pollution. Lead is a major source of neurotoxin that can induce neural deformities. Lactobacillusspecies isolated from curd were characterized to confirm its specificity. Zebra fish was reared at standard conditions and preclinical assessment on the intensity of induced neurotoxin lead was performed. The embryo toxic assay, immunomodulation effects and animal behavioural models endorsed the consequence of neurotoxicity. Different concentrations of bacterial isolate with standard antidepressant was considered for analysing the vigour of toxicity and its influence on cognitive behaviour by novel tank diving method. The restrain in the animal behaviour was also conferred by all the test samples with a decreased bottom dwelling time which was authenticated with haematology and histopathological studies. The alterations in morphology of the lymphocytes were balanced by the treated test samples. This study paves a twofold potential of probiotic as neuroprotectant and immune modulator against heavy metal toxicity.

Caracterização fenotípica e funcional das células T CD4+ e CD8+ VB12 na resposta imune em paciente com Leishmaniose Cutânea Localizada / Phenotypic and functional characterization of CD4 + and CD8 + VB12 T cells in the immune response in a patient with Localized Cutaneous Leishmaniasis

As leishmanioses constituem um complexo de doenças causada pelo protozoário intracelular, do gênero Leishmania, sendo a resposta imune celular essencial para controle, eliminação e proteção contra a infecção. A teoria clonal da imunidade celular propõe que as respostas imunológicas são estabelecidas através do aumento na frequência de clones específicos ao antígeno. Para avaliar a resposta das células T à infecção por Leishmania, investigamos, por citometria de fluxo, a expressão de cadeias Vβ de receptores de células T (TCRs), estado de ativação, capacidade de adesão ao endotélio e potencial funcional de clones específico. Em um grupo de pacientes com Leishmaniose Cutânea Localizada (LCL), avaliamos diferentes subpopulações de células T através da expressão da região Vβ, no sangue periférico e na biópsia da lesão. Utilizamos células mononucleares de sangue periférico (CMSPs), de pacientes LCL e controles saudáveis, nas quais avaliamos, ex vivo, a expressão de moléculas de ativação (CD25, CD69 e HLA-DR), adesão (LFA-1, VLA-4 e CD62L), co-estimulatória (CD27 e CD28)...

Leishmaniasis is a desease caused by infection with the Leishmania protozoan parasite. The cellular immune response is essential for controlling, eliminating and protection of the Leishmania infection. The clonal theory of cellular immunity proposes that immunological responses are established by increasing the frequency of antigen-specific clones. In order to measure the host T cell response to Leishmania infection, we have investigated by flow cytometry, the expression of Vβ chains of T-cell receptors (TCRs), activation state, adhesion to endothelium of capacity and functional potential of specific T. In a group of localized cutaneous leishmaniasis (LCL) patients, we evaluated different T cell subpopulations as identified by their Vβ region expression, in peripheral blood and biopsy. We used peripheral blood mononuclear cells (PBMCs), from CL patients and healthy volunteers, in which we evaluate, ex vivo, the expression of activation molecules (CD25, CD69 and HLA-DR), adhesion (LFA-1, VLA-4 and CD62L), co-stimulatory (CD27 and CD28)...

Efecto in vitro de la Matricaria recutita L. sobre la respuesta de linfocitos y neutrófilos / In vitro effect of Matricaria recutita L. on Lymphocytes and Neutrophils response

La manzanilla de Castilla, dulce o cimarrona (Matricaria recutita o Matricaria chamomilla), es una planta herbácea anual de la familia de las asteráceas, nativa de Europa y de regiones templadas de Asia, que se ha naturalizado en algunas regiones de América, África y Australia. Ha sido utilizada por el hombre desde hace miles de años con diferentes fines medicinales. Se estudió el efecto in vitro de un extracto fluido de esta planta sobre los linfocitos de 20 donantes voluntarios de sangre y de 20 enfermos con diagnóstico de inmunodeficiencia celular, mediante la cuantificación de los linfocitos T por las técnicas de formación de roseta espontánea y activa y el ultramicrométodo inmunocitoquímico (UMICIQ), así como la función fagocítica (índice opsonofagocítico) de los neutrófilos. No se hallaron diferencias estadísticamente significativas en los parámetros estudiados entre las condiciones experimentales con Matricaria y sin esta

Castilla Chamomile, sweet, or maroon (Matricaria recutita or Matricaria chamomilla) is an annual herbaceous plant of the Asteraceae family, native to Europe and temperate zones of Asia, which has become naturalized in parts of America, Africa, and Australia. It has been used by man for thousands of years with different medicinal purposes. We studied the in vitro effect of an extract fluid of this plant on lymphocytes from 20 blood donors and 20 patients with a diagnosis of cellular immunodeficiency. We quantified T lymphocytes by the techniques of spontaneous and active rosette formation, and the immunocytochemical ultramicromethod (UMICIQ) as well as the phagocytic function (opsonophagocytic index) of neutrophils. There were no statistically significant differences in the studied parameters between experimental conditions with Matricaria and without it

Clastogenic effects of different Ureaplasma urealyticum serovars on human chromosomes

The possibility that Ureaplasma urealyticum might play an important role in human infertility was first raised more than 20 years ago, but this association remains speculative. Considering the hypothesis that the pathogenicity of Ureaplasma urealyticum may depend on its serotypes, the clastogenic effcts of different strains of Ureaplasma urealyticum, at concentrations of 10(3) CCU (color changing units)/ml, 10(4) CCU/ml and 10(5) CCU/ml, were evaluated in vitro in short-term cultures of human lyphocytes. Total or partial mitotic inhibition was produced by Ureaplasma urealyticum serotypes 2,3 and 10 independent of the concentration (10(3) CCU/ml, 10(4) CCU/ml or 10 (5) CCU/ml) of the microorganisms employed. In contrast, the clastogenic effects observed with serotypes 1,7 and 12 varied according to the concentration employed in the test. Mitotic alterations were observed in Ureaplasma urealyticum serotypes 5,6,7,8,9,11 and 12. Chromatid gaps (53.0 percent) and chromatid breaks (13.9 percent) were the most frequent types of alterations observed. The results of this in vitro assay demonstrated that the clastogenic effects varied with the Ureaplasma urealyticum serotypes evaluated.

Modificación de un método colorimétrico que usa XTT, para la determinación de linfoproliferación / Modification of a colorimetric method using XTT, for determination of lymphoproliferation

Los métodos para la determinación de proliferación celular se han basado tradicionalmente en la incorporación y detección de productos marcados con radioactividad. En este trabajo se describe una modificación de una técnica colorimétrica capaz de detectar la linfoproliferación. El estudio se llevó a cabo con linfocitos de 23 estudiantes universitarios voluntarios aparentemente sanos y se obtuvieron diferentes significativas (Tt>Tc) entre linfocitos estimulados y no estimulados. El método permite cuantificar la linfoproliferación por medio de una medición espectrofotométrica directa en un lector de ELISA. El número de células presente en cada pozo correlaciona con la capacidad de reducir la sal de tetrazol (XTT) a un producto coloreado y soluble. La sal es reducida principalmente por la actividad enzimática mitocondrial presente básicamente en las células vivas. La técnica presenta las ventajas de ser simple, económica y segura si se compara con los métodos que emplean radioactividad comúnmente usados; esto podría facilitar su introducción en Costa Rica, en los laboratorios de investigación y en el área hospitalaria, para el apoyo en el diagnóstico de problemas del sistema inmune.

Hairy cell transformation of human peripheral blood lymphocytes by Coxiella burnetii

Hairy cell (HC) transformation of human peripheral blood lymphocytes (PBL) by Coxiella burnetii was studied to clarify the significance of persistency of C. burnetii in a hairy cell line (designated "TOL"). TOL cells which exhibited HC characteristics in hairy cell leukemia (HCL) were persistently infected with C. burnetii. Two strains of C. burnetii, our isolate from TOL cells and the original isolate in 1935, the Nine Mile strain from American Type Culture Collection (ATCC, U.S.A), were inoculated to PBL cultures. HC transformation not only by our isolates (87%) but also by Nine Mile strain (100%) was demonstrated in an average of 20 days. The original observation that Coxiella induced HC transformation in vitro was also confirmed in experiments with PBL exposed to C. burnetii in vivo. Spontaneous development of HC were observed in cultures of PBL only from coxiellemic cases (12/24) but not from C. burnetii negative cases (0/57). All HC cell lines (34) as determined by their morphology and cytochemical markers of HC in HCL remained infected with C. burnetii invariably.

Estudio al microscopio electrónico de los linfocitos en la sangre periférica de pacientes epilépticos / Electronmicroscope study of lymphocites in the peripheral blood of epileptic patients

Se tratan de establecer los probables mecanismos inmunológicos involucrados en la patogenia de la epilepsia y por ello se investigaron los linfocitos de pacientes con epilepsia de etiología desconocida, con y sin tratamiento para estudiar el grado de diferenciación celular linfocitaria. Se aislaron las poblaciones de células mononucleares de sangre periférica por centrifugación en gradiente de densidad, de Ficoll-Hypaque de acuerdo a la técnica de Boyun. Se estudiaron 22 pacientes, de los cuales 16 (72.73 por ciento) habían recibido tratamiento con medicamento anticonvulsivante, y 6 (27.27 por ciento) estaban sin tratamiento. Se tomaron además 10 controles sanos. Las fracciones de células aisladas se procesaron por las técnicas convencionales para microscopia electrónica de transmisión. El estudio de las micrografías electrónicas de los controles mostró 65 por ciento de linfocitos en reposo, 22.5 por ciento de linfocitos activados y 12.13 por ciento degenerados. En los pacientes epilépticos de etiología desconocida, tratados y no tratados, se observó un alto porcentaje de linfocitos activados. 89.44 por ciento y 73.33 por ciento respectivamente. Esta activación consistió en linfocitos con predominio de ribosomas libres, con desarrollo del retículo endoplásmico rugoso (RER) y liso plasmocitoide. En los pacientes epilépticos con tratamiento y de etiología conocida se observaron linfocitos en reposo, linfocitos activados con aumento de ribosomas libres y con desarrollo del RER, pero se observaron linfocitos plasmocitoides. El aumento del retículo endoplásmico liso encontrado en algunos linfocitos de pacientes epilépticos con tratamiento pudiera atribuirse al tratamiento con fenobarbital. La difenihidaltonia parece inducir cambios degenerativos