Subpoblaciones de linfocitos T en adultos mayores cubanos con leucemia linfoide crónica / Subpopulations of T lymphocytes in Cuban older adults with chronic lymphoid leukemia

Introducción: la inmunosenescencia está asociada con un mayor riesgo de desarrollo de cáncer. Dentro de las hemopatías malignas que afectan a este grupo de edad, está la leucemia linfoide crónica (LLC), caracterizada por trastornos en la inmunidad adaptativa que incluye las subpoblaciones de linfocitos T. Objetivo: Determinar la frecuencia de las subpoblaciones de linfocitos T en los pacientes adultos mayores con leucemia linfoide crónica evaluados en el Instituto de Hematología e Inmunología de Cuba. Métodos: Se realizó un estudio transversal en 30 adultos mayores con leucemia linfoide crónica. Se cuantificaron los linfocitos TCD3+CD4+ y TCD3+CD8+ en sangre periférica por citometría de flujo. Para la lectura y el análisis de los datos se empleó un citómetro de flujo Beckman Coulter Gallios. Se utilizaron los valores porcentuales, la media y la desviación estándar. Se consideró estadísticamente significativo si p≤0.05. Resultados: Hubo un predominio de hombres que representaron el 56,7 por ciento y del grupo de 70-79 años de edad. No se reportó ningún adulto mayor con LLC con valores altos ni normales de linfocitos TCD3+CD4+. Predominaron los hombres con valores bajos porcentuales de linfocitos TCD3+CD4+, TCD3+CD8+ e inversión del índice CD4/CD8 en relación con las mujeres. Conclusiones: Los adultos mayores con LLC presentan alteraciones en el número de las subpoblaciones de linfocitos T. La acción de estas células en relación al crecimiento de células B malignas aún es desconocido y resulta importante determinar si esto puede reflejar un intento de evasión de las células tumorales al control inmunológico(AU)

Introduction: Immunosenescence is associated with an increased risk of cancer development. Among the malignant hemopathies that affect this age group, it is chronic lymphoid leukemia (CLL), characterized by disorders in adaptive immunity, which include subpopulations of T lymphocytes. Objective: To determine frequency of T lymphocyte subpopulations in older adult patients with chronic lymphoid leukemia evaluated at the Institute of Hematology and Immunology of Cuba. Methods: A cross-sectional study was conducted in 30 older adults with chronic lymphoid leukemia. TCD3+CD4+ and TCD3+CD8+ lymphocytes were quantified in peripheral blood by flow cytometry. A Beckman Coulter Gallios flow cytometer was used to read and analyze the data. The percentage values, the mean and the standard deviation were used. It was considered statistically significant if p≤0.05. Results: There was a predominance of men who represented 56.7 percent and the age group of 70-79 years. No older adults with CLL with high or normal values of TCD3+CD4+ lymphocytes were reported. Men predominated with low percentage values of TCD3+CD4+, TCD3+CD8+ lymphocytes and inversion of the CD4/CD8 ratio in relation to women. Conclusions: Older adult with CLL present alterations in the number of T lymphocyte subpopulations. The role of these cells in relation to the growth of malignant B cells it is unknown and it turns out important to determine if this may reflect an attempt to evade tumor cells from immune control(AU)

Immune reconstitution after allogenic stem cell transplantation: An observational study in pediatric patients

Abstract Introduction The immune reconstitution (IR) after the allogenic hematopoietic stem cell transplantation (allo-HSCT) is a progressive process intrinsically correlated to the therapeutic success. It is essential to understand the interfering factors in IR to prevent the HSCT-related mortality. Methods We retrospectively evaluated the clinical outcomes, absolute lymphocyte counts (ALCs) and lymphocyte subtypes at different time-points of 111 pediatric patients with allogeneic HSCT for malignant and non-malignant diseases from 2013 to 2018. Results The ALCs gradually increased on D+30, D+100, and D+180 (medians 634/μL, 1022/μL and 1541/μL, respectively). On D+100, the CD3+CD8+ achieved the highest recovery rate (68%), followed by the CD16+CD56+ (47%), CD3+CD4+ (39%) and CD19+ (8%). The adequate ALC recovery was associated with age < 8 years, bone marrow grafts, myeloablative conditioning, non-use of serotherapy and non-haploidentical donors. The ALC and CD3+CD8+ on D+100 counts were higher in patients with the cytomegalovirus infection. The CD3+CD4+ recovery was associated with an age < 8 years, a non-malignant disease and a lower incidence of acute graft-versus-host disease ≥ grade 2. Furthermore, the ALC recovery on D+100 resulted in a higher overall survival, regardless of the disease type (HR 3.65, 1.05 - 12.71, p= 0.04). Conclusion Several factors influenced the IR after the allo-HSCT. The ALC ≥ 500/μL on D+100 was a simple IR predictor of survival, easily available to resource-limited centers.

The Relationship between Occurrence of aGVHD in Patients with Acute Myeloid Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation and Immune Cell Components in Graft / 中国实验血液学杂志

OBJECTIVE@#To explore the relationship between occurrence of acute graft-versus-host disease (aGVHD) and various immune cell composition in patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*METHODS@#The clinical data of 104 patients with AML undergoing allo-HSCT in our hospital were retrospectively analyzed, and the hematopoietic reconstitution and occurrence of GVHD were analyzed. Flow cytometry was used to detect the proportion of various types of immune cells in the grafts, the number of graft composition in patients with different degrees of aGVHD was calculated and compared, and to analyze the correlation between the severity of aGVHD in AML patients after allo-HSCT and the immune cell components in the graft.@*RESULTS@#There was no significant difference in the time of hematopoietic reconstitution between the high number group of total number of nucleated cells (TNC) and the low number group, while the time of neutrophil and platelet reconstruction in the high number of CD34 group was significantly faster than that in the low number of CD34 group (P＜0.05), and the total hospital stay also tends to be shorten. Compared with patients in 0-Ι aGVHD group, both HLA-matched and HLA-haploidentical transplantation, the infusion amounts of CD3+ cells, CD3+CD4+ cells, CD3+CD8+ cells, NK cells and CD14+ monocytes were higher in patients of Ⅱ-Ⅳ aGVHD group， but the difference was not statistically significant (P>0.05); In addition, in patients with HLA-haploidentical transplantation, the number of CD4+CD25+ cells in Ⅱ-Ⅳ aGVHD group was significantly lower than that in 0-Ι aGVHD group (P<0.05), and the same trend was also observed in HLA-matched transplanted patients, but the difference was not significant (P=0.078).@*CONCLUSION@#High number of CD34+ cells in the graft is beneficial to hematopoietic reconstitution in AML patients. To a certain degree, high number of CD3+ cells, CD3+CD4+ cells, CD3+CD8+ cells, NK cells and CD14+ cells tend to increase the occurrence of aGVHD, but high number of CD4+CD25+ regulatory T cells is beneficial to reduce the incidence of aGVHD in AML patients.

Avances en la inmunopatogenia de la hepatitis autoinmune / Advances in the immunopathogenesis of autoimmune hepatitis

RESUMEN Introducción: en la hepatitis autoinmune los mecanismos inmunopatogénicos no están totalmente esclarecidos, múltiples son las investigaciones en este campo, con vistas a enriquecer los conocimientos y ampliar las opciones terapéuticas. Objetivo: sintetizar los conocimientos más recientes acerca de la inmunopatogenia de esta enfermedad. Material y Método: se efectúa una búsqueda exhaustiva de la bibliografía disponible en SciELO, ScienceDirect, Google Académico y PubMed, incluyendo artículos de revisión, estudios experimentales, clínicos, de cohorte y metaanálisis. Desarrollo: se explican los principales mecanismos de tolerancia central y periférica, así como el papel de las subpoblaciones linfoides, las citocinas y el microambiente en la patogenia de la enfermedad. Conclusiones: los avances en el conocimiento de la inmunopatogenia de la hepatitis autoinmune permiten una mejor comprensión de esta enfermedad y son el referente para el diseño de estrategias futuras de tratamiento.

ABSTRACT Introduction: Immunopathogenic mechanisms are not fully clarified in autoimmune hepatitis; there are many investigations in this field with a view to enriching knowledge and expanding therapeutic options. Objective: To synthesize the most recent knowledge about the immunopathogenesis of this disease. Material and Methods: An exhaustive search of the bibliography available in SciELO, ScienceDirect, Google Scholar and PubMed was carried out, including review articles, experimental, clinical, cohort and meta-analysis studies. Development: The main mechanisms of central and peripheral tolerance are explained, as well as the role of lymphoid subpopulations, cytokines and the microenvironment in the pathogenesis of the disease. Conclusions: Advances in the knowledge of the immunopathogenesis of autoimmune hepatitis allow a better understanding of this disease and are the referents in the design of future treatment strategies.

Determinación del intervalo de referencia para subpoblaciones linfocitarias CD3+, CD4+ y CD8+ y el índice CD4+/CD8+ en la población chilena adulta

Resumen Introducción: En varios estudios se ha documentado la influencia de factores étnicos en la distribución de las subpoblaciones linfocitarias; sin embargo, los intervalos de referencia utilizados en Chile fueron obtenidos de un estudio realizado en Países Bajos el año 1997. Objetivo: Determinar el intervalo de referencia para subpoblaciones linfocitarias CD3+, CD4+ y CD8+, además del índice CD4+/CD8+ en la población chilena. Metodología: Se analizó un total de 200 muestras de sangre total obtenida de hombres y mujeres adultos sanos, utilizando el método establecido por el CLSI estandarizado en el protocolo EP28-A3c desde la etapa pre analítica en adelante. Resultados: Los rangos de referencia para CD3+, CD4+ y CD8+ fueron 54,7-81,6% (789-2732 céls/μL), 28,1-57,7% (447-1703 céls/μL) y 15,1-38,8% (226-996 céls/μL), respectivamente. El índice CD4+/CD8+ fue de 0,84-3,77. Discusión: Los valores de referencia de las subpoblaciones linfocitarias en la población chilena sana son diferentes de los que se usan actualmente en Chile. Estas observaciones muestran datos locales que pudieran tener implicaciones para el tratamiento de la infección por VIH, y los rangos de referencia encontrados en este estudio pudieran ser usados para entender la situación local de algunos pacientes. Conclusiones: Otros estudios deberán ser realizados para confirmar estas observaciones dada la falta de datos previos y debido a que este es el primer estudio en población chilena.

Abstract Background: Several studies have documented the influence of ethnic factors on the distribution of lymphocyte subpopulations; however, the reference intervals used in Chile were obtained from a study carried out in the Netherlands in 1997. Aim: To determine the reference interval for CD3+, CD4+ and CD8+ lymphocyte subpopulations, as well as the CD4+/CD8+ index in the Chilean population. Methods: A total of 200 whole blood samples obtained from healthy adult men and women were analyzed using the method established by CLSI standardized in protocol EP28-A3c from the pre-analytical stage onwards. Results: The reference ranges for CD3+, CD4+ and CD8+ were 54.7-81.6% (789-2732 cells/μL), 28.1-57.7% (447-1703 cells/μL) and 15.1-38.8% (226-996 cells/μL), respectively. The CD4+/ CD8+ index was 0.84-3.77. Discussion: The reference values for lymphocyte subpopulations in the healthy Chilean population are different from those currently used in Chile. These observations show local data that could have implications for the treatment of HIV infection and the reference ranges found in this study could be used to understand the local situation of some patients. Conclusions: Others studies must be done to confirm these observations due to lack of previous data and because this is the first study in Chilean population.

Alteração na representação de subpopulações de neurônios no epitélio olfatório de camundongos Ric8b cKO / Altered representation of neuronal sub-populations in the olfactory epithelium of Ric8b conditional knock-out mice

O objetivo desse trabalho foi identificar as consequências moleculares e funcionais da falta da proteína Ric8b no epitélio olfatório de camundongos. Para esse fim, comparamos o transcriptoma de epitélio olfatório de camundongos knock-out tecido específico para a proteína RIC8B (Ric8b cKO) com o dos seus irmãos tipo selvagem (WT). Identificamos muitos genes que apresentaram expressão reduzida no epitélio olfatório do camundongo Ric8b cKO, mas também vários genes que apresentaram a sua expressão aumentada. A maioria dos genes com expressão reduzida corresponde a genes normalmente expressos em neurônios olfatórios maduros, como por exemplo os genes de receptores olfatórios, o que é compatível com o fato já conhecido de que os camundongos Ric8b cKO apresentam um menor número desses neurônios. Inesperadamente, apesar de a maioria dos genes de receptores olfatórios ter a sua expressão diminuída no camundongo Ric8b cKO, observamos que um grupo destes genes de receptores teve a sua expressão aumentada. Os camundongos Ric8b cKO apresentaram também genes marcadores de outros tipos celulares que não neurônios canônicos com expressão aumentada no seu epitélio olfatório. Dentre eles, os mais significativamente alterados foram os genes marcadores de neurônios Trpc2+ tipo B (que expressam a guanilato ciclase solúvel Gucy1b2). Sabe-se que este tipo de neurônio é responsável pela sensibilidade a diferentes gases, e concordantemente, observamos que os camundongos Ric8b cKO apresentaram um aumento da sensibilidade a gás carbônico. Como o olfato apresenta um papel importante na regulação de ingestão alimentar, analisamos como os camundongos Ric8b cKO se comportam frente a diferentes dietas. Interessantemente, observamos que esses animais não apresentam preferência por alimento rico em gorduras quando comparado aos seus irmãos tipo selvagem. Nossos resultados sugerem, portanto, que a ausência da proteína RIC8B resulta na alteração de representatividade de neurônios canônicos e não canônicos no epitélio olfatório de camundongos, o que por sua vez leva a alterações funcionais e comportamentais

The objective of this work was to identify the molecular and functional consequences of the lack of the RIC8B protein in the main olfactory epithelium of mice. To this end, we compared the olfactory epithelium transcriptome of Ric8b tissue-specific knock-out mice (Ric8b cKO) with that of their wild-type littermates (WT). We identified many genes with differential expression, many of which were downregulated and also some which were upregulated in the olfactory epithelium of the Ric8b cKO mice. Most of the downregulated genes correspond to genes normally expressed in mature olfactory sensory neurons, such as olfactory receptor genes. This is compatible with the already known fact that the Ric8b cKO mice have less of this kind of neuron. Unexpectedly, even though most of the olfactory receptor genes were downregulated, we observed a subset of these genes that had their expression upregulated in the Ric8b cKO mice. The Ric8b cKO mice also showed upregulation for genes that are markers for cell types other than canonic neurons in their olfactory epithelium. Among these, the most significantly altered were the markers for neurons Trpc2+ type B (that express the soluble guanylate cyclase Gucy1b2). It is known that this kind of neuron is responsible for sensitivity to different gases. Accordingly, we observed that the Ric8b cKO mice presented a higher sensitivity to carbon dioxide. Since olfaction has an important role in food intake, we analyzed how the Ric8b cKO mice behaved with different diets. Interestingly, we observed that the Ric8b cKO mice lack preference for high fat diet when compared to their wild-type littermates. Our results indicate, therefore, that the lack of the RIC8B protein results in altered representativity of canonic and non-canonic neurons in the olfactory epithelium of mice, which then leads to altered function and behavior

Back to Tityus serrulatus Lutz & Mello, 1922 (Scorpiones: Buthidae): new comments about an old species

A synopsis on the historical, geographical and ecological aspects related to the most conspicuous scorpion species of the genus Tityus known from Brazil is proposed. Tityus serrulatus Lutz & Mello, 1922 was described precisely one century ago, nevertheless many questions related to its ecological adaptations and geographical expansion remain without a precise response. This species, well known for its infamous reputation of noxious species, is also known for its capacity to reproduce asexually, by parthenogenesis. Although the individuals of a given population are considered clones, a new hypothesis could suggest the occurrence of mutations within isolated individuals, leading to distinct subpopulations that could present better phenotypic performances in ecological habitats distinct from those of the original area of distribution of the species.(AU)

Flow cytometry evaluation of CD14/CD16 monocyte subpopulations in systemic sclerosis patients: a cross sectional controlled study

Abstract Background: Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by vasculopathy and fibrosis, which can be subclassified into diffuse cutaneous (dSSc) and limited cutaneous (lSSc) subtypes. Previous studies suggest that an increase in monocytes can be a hallmark of various inflammatory diseases, including SSc. Our aim was to evaluate circulating blood monocyte subpopulations (classical, intermediate and non-classical) of SSc patients and their possible association with disease manifestations. Methods: Fifty consecutive patients fulfilling the 2013 ACR/EULAR classification criteria for SSc were included in a cross-sectional study. Monocyte subpopulations were identified based on their expression of CD64, CD14 and CD16, evaluated by flow cytometry, and were correlated with the clinical characteristics of the patients; furthermore, the expression of HLA-DR, CD163, CD169 and CD206 in the monocytes was studied. Thirty-eight age- and sex-matched healthy individuals were recruited as a control group. Results: SSc patients had an increased number of circulating peripheral blood monocytes with an activated phenotypic profile compared to healthy subjects. Absolute counts of CD16+ (intermediary and non-classical) monocyte subpopulations were higher in SSc patients. There was no association between monocyte subpopulations and the clinical manifestations evaluated. Conclusion: We identified higher counts of all monocyte subpopulations in SSc patients compared to the control group. There was no association between monocyte subpopulations and major fibrotic manifestations. CD169 was shown to be more representative in dSSc, being a promising marker for differentiating disease subtypes.

Distribution of peripheral blood lymphocyte subpopulations in 654 children of preschool age / 国际儿科学杂志

Objective@#To investigate the distribution of peripheral blood lymphocyte subpopulations in 654 children aged 28 days to 7 years, and to provide a basis for establishing a normal reference range.@*Methods@#A total of 654 healthy Han children aged 28 days to 7 years were enrolled.The children were divided into infant group(28 days-12 months)(180 cases, 27.52%), toddler group(1-3 years)(184 cases, 28.13%), and preschooler group(3-7 years)(290 cases, 44.34%). Peripheral blood samples were collected, and the percentages of lymphocyte subpopulation were detected by flow cytometry.@*Results@#There were statistically significant differences between boys and girls in CD3+ CD4+ T cells, CD3+ CD8+ T cells percentages and the CD4+ /CD8+ ratio in infant group(Z=-2.595, 3.317, -3.492, all P<0.05); in CD3+ CD4+ T cells percentage in toddler group(Z=2.312, P<0.05); in CD3+ T cells, CD3+ CD4+ T cells, CD3-CD19+ B cells, CD3-CD16+ CD56+ NK cells percentages and the CD4+ /CD8+ ratio in preschool age group(Z=4.088, 4.991, 3.129, -6.949, 2.141, all P<0.05). The comparison in all age groups showed significant differences in CD3+ T cells, CD3+ CD4+ T cells, CD3-CD19+ B cells, CD3-CD16+ CD56+ NK cells percentages and the CD4+ /CD8+ ratio in boys(χ2=6.925, 51.543, 39.563, 87.751, 30.334, all P<0.05), in CD3+ CD4+ T cells, CD3+ CD8+ T cells, CD3-CD16+ CD56+ NK cells percentages and the CD4+ /CD8+ ratio in girls(χ2=27.646, 44.046, 26.066, 54.238, all P<0.05). The CD3+ CD4+ T cells and CD3-CD19+ B cells percentages declined with age(χ2=82.345, 40.214, all P<0.05); The CD3+ CD8+ T cells and CD3-CD16+ CD56+ NK cells percentages increased with age(χ2=38.43, 108.302, all P<0.05).@*Conclusion@#The peripheral blood lymphocyte subpopulation values differ by gender and age.It is necessary to establish the reference range of lymphocyte subpopulations for children in Lanzhou according to gender and age.

Distribution of peripheral blood lymphocyte subpopulations in 654 children of preschool age / 国际儿科学杂志

Objective To investigate the distribution of peripheral blood lymphocyte subpopulations in 654 children aged 28 days to 7 years,and to provide a basis for establishing a normal reference range.Methods A total of 654 healthy Han children aged 28 days to 7 years were enrolled.The children were divided into infant group (28 days-12 months) (180 cases,27.52％),toddler group (1-3 years) (184 cases,28.13 ％),and preschooler group (3-7 years) (290 cases,44.34％).Peripheral blood samples were collected,and the percentages of lymphocyte subpopulation were detected by flow cytometry.Results There were statistically significant differences between boys and girls in CD3+ CD4 + T cells,CD3 + CD8 + T cells percentages and the CD4 +/CD8 + ratio in infant group (Z =-2.595,3.317,-3.492,all P ＜ 0.05);in CD3 +CD4 + T cells percentage in toddler group (Z =2.312,P ＜ 0.05);in CD3 + T cells,CD3 + CD4 + T cells,CD3-CD19 + B cells,CD3-CD16 + CD56 + NK cells percentages and the CD4 +/CD8 + ratio in preschool age group (Z =4.088,4.991,3.129,-6.949,2.141,all P ＜ 0.05).The comparison in all age groups showed significant differences in CD3 + T cells,CD3 + CD4 + T cells,CD3-CD19 + B cells,CD3-CD16 +CD56 + NK cells percentages and the CD4 +/CD8 + ratio in boys (x2 =6.925,51.543,39.563,87.751,30.334,all P ＜ 0.05),in CD3 + CD4 + T cells,CD3 + CD8 + T cells,CD3-CD16 + CD56 + NK cells percentages and the CD4 +/CD8 + ratio in girls (x2 =27.646,44.046,26.066,54.238,all P ＜ 0.05).The CD3 +CD4+ T cells and CD3-CD19+ B cells percentages declined with age (x2 =82.345,40.214,all P ＜0.05);The CD3+ CD8+ T cells and CD3-CD16+ CD56+ NK cells percentages increased with age (x2 =38.43,108.302,all P ＜0.05).Conclusion The peripheral blood lymphocyte subpopulation values differ by gender and age.It is necessary to establish the reference range of lymphocyte subpopulations for children in Lanzhou according to gender and age.

Favorable responses to treatment with 5 mg Sbv/kg/day meglumine antimoniate in patients with American tegumentary leishmaniasis acquired in different Brazilian regions

Abstract INTRODUCTION: Favorable responses in American tegumentary leishmaniasis (ATL) patients to treatment with 5 mg Sbv/kg/day meglumine antimoniate (MA) has been reported in Rio de Janeiro, but little is known regarding the therapeutic response to low doses in patients from other locations. METHODS: A retrospective review of medical records was conducted to compare the therapeutic response to 5 mg Sbv/kg/day MA treatment among 36 patients who acquired ATL in Brazilian states other than Rio de Janeiro (OS group) and 72 patients from Rio de Janeiro (RJ group). RESULTS: One course of 5 mg Sbv/kg/day MA cured 72.8% of 81 cutaneous (CL) and 66.6% of 27 mucosal (ML) leishmaniasis-infected patients: 70% in the CL/RJ group, 81% in the CL/OS group, 50% in the ML/RJ group, and 80% in the ML/OS group. After up to two additional treatment courses at the same dose, 88.9% and 85.2% of the CL and ML patients were cured, respectively. Adverse events were observed in 40% of patients in the CL/RJ group, 57% of the CL/OS group, 58% of the ML/RJ group, and 80% of the ML/OS group. No significant differences were observed in the cure rates or adverse effects between the RJ and OS groups. No patients required permanent discontinuation of treatment due to adverse events. CONCLUSIONS: Patients with ATL acquired in both RJ and OS may respond to low-dose MA. While high-dose MA should remain the standard treatment for ATL, low-dose MA might be preferred when toxicity is a primary concern.

Comparison of T-lymphocyte subpopulations,complement C3,C4 and HBV-DNA in theperipheral blood of patients with chronic hepatitis B under different hepatic function / 国际检验医学杂志

Objective To compare the T-lymphocyte subpopulations,complement C3,C4 and HBV-DNA in the peripheral blood of patients with chronic hepatitis B under different liver function.Methods A total of 136 patients with chronic hepatitis B were selected,the patients were divided into improved group and deteriorated group according to the changes of hepatic function.T-lymphocyte subpopulations,complement C3,complement C4 and HBV-DNA in two groups were determined.Results A total of 72 patients were included in improved group,64 patients were included in deteriorated group.CD4+ in deteriorated group was significantly decreased,and CD8+ was significantly increased compared with those in improved group(P0.05).Complement C3 and complement C4 in deteriorated group decreased compared with those in the improved group(t=12.124,P=0.003;t=4.041,P=0.010).However,HBV-DNA had no statistical difference between two groups(t=-2.598,P=0.793 ).Conclusion Compared with T-lymphocyte subpopulations,complement C4 and HBV-DNA,complement C3 has a better sensitivity to reflect the damage of the hepatic function in patients with chronic hepatitis B.

Effect of angiotensin II on phenotypic transformation in vascular adventi-tial fibroblast subpopulations / 中国病理生理杂志

AIM:To explore the effect of angiotensin II ( Ang II) on phenotypic transformation in adventitial fibroblast subpopulations isolated from rat thoracic aorta .METHODS: Vascular adventitial fibroblasts were individually expanded by the method of cloning rings .The isolated cells were identified by reverse transcription-polymerase chain reac-tion ( RT-PCR) and immunofluorescence staining .The rat vascular adventitial fibroblasts were cultured in vitro and incuba-ted with or without Ang II.Using the methods of fluorescence quantitative polymerase chain reaction (FQ-PCR), immuno-fluorescence staining and Western blot , the expression of α-smooth muscle actin (α-SMA) was detected .RESULTS:The spindle cells and round cells were isolated by the method of cloning rings .From passage 3 to passage 8,α-SMA expression was decreased in the spindle cells , and increased significantly in the round cells .Induced by Ang II , the expression of α-SMA was increased in the 2 cell subpopulations, and that in the round cells was significantly increased (P<0.01).The results of Western blot indicated that Ang II stimulated the subpopulation of the round cells to transfer to myofibroblasts . CONCLUSION:Ang II promotes the phenotypic transformation of adventitial fibroblast subpopulations , and the 2 subpop-ulations may play an important role in vascular remodeling and reparative process .

Sperm subpopulations in avian species: A comparative study between the rooster (Gallus domesticus) and Guinea fowl (Numida meleagris) / 亚洲男科学杂志(英文版)

The main aims of this research were to study possible differences in objective morphometric sperm characteristics, establish normative sperm morphometry standards, and evaluate the presumed different subpopulation distribution of avian spermatozoa from the rooster (Gallus domesticus ) and Guinea fowl (Numida meleagris ) as model avian species. Seventy-two ejaculates (36 per species studied) were obtained manually, following a training period involving gently combined dorso-abdominal and lumbo-sacral massage of the birds. Ejaculates were processed for volume, sperm concentration, viability, motility, and morphology. Moreover, samples were submitted for sperm morphometric assessment using objective Computer-Assisted Semen Analysis for Morphometry (CASA-Morph) methods, with sperm morphometric descriptors evaluated by Principal Component Analysis (PCA) and multivariate clustering analyses. There were several differences observed between the avian species in values obtained for ejaculate volume and sperm concentration (P < 0.001). Irrespective of species, PCA revealed two Principal Components (PCs) explaining more than 80% of the variance. In addition, the number of subpopulations differed with species (three and five subpopulations for rooster and Guinea fowl, respectively). Moreover, the distribution of the sperm subpopulations was found to be structurally different between species. In conclusion, our findings from using CASA-Morph methods indicate pronounced sperm morphometric variation between these two avian species. Because of the strong differences observed in morphometric parameter values and their subpopulation distribution, these results suggest that application of objective analytical methods such as CASA-Morph could substantially improve the reliability of comparative studies and help establish valid normative sperm morphological values for avian species.

Normozoospermic versus teratozoospermic domestic cats: Differential testicular volume, sperm morphometry, and subpopulation structure during epididymal maturation / 亚洲男科学杂志(英文版)

Teratozoospermia (<40% morphologically normal spermatozoa/ejaculate) is a frequent phenomenon in feline species. This research was carried out to study the possible differences in testicular volume, differential sperm morphometric traits, and potential differences regarding the sperm subpopulational structure during epididymal sperm maturation in teratozoospermic feline donors. Epididymal sperm samples were collected from the caput (R1), corpus (R2), and cauda (R3) epididymidis in two donor groups (N: normozoospermic; T: teratozoospermic). Aliquots were assessed for concentration, viability, motility, and acrosomal integrity. Sperm morphometric descriptors from CASA-Morph analysis were analyzed by the Principal Component Analysis (PCA) and clustering analyses. Irrespective of the group analyzed, PCA revealed two Principal Components (PCs) for each epididymal region explaining more than the 93% of the variance. Surprisingly, the number of subpopulations remained constant in regions R1-R2-R3 irrespective of the donor group analyzed. However, the distribution of these subpopulations was found to be structurally different and strongly influenced by the epididymal region and the donor group. In conclusion, testicular morphometry and the sperm subpopulation structure were different in N and T donors. The alterations in subpopulations during epididymal maturation could be used as a potential clinical indicator of teratozoospermic individuals since an important influence of teratozoospermia on sperm subpopulation structure has been demonstrated.

Morphometry and subpopulation structure of Holstein bull spermatozoa: Variations in ejaculates and cryopreservation straws / 亚洲男科学杂志(英文版)

Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the «classical» statistical approach, based on variance analysis (revealing differences between animals, ejaculates, and straws), principal component (PC) analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.

Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men / 亚洲男科学杂志(英文版)

This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis (CASA-Mot), and of sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations (slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]), and three morphometric subpopulations (large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]). The distribution of kinematic sperm subpopulations was different among ejaculate fractions (P < 0.001), with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions (P < 0.001), with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

Comparison of different statistical approaches to evaluate morphometric sperm subpopulations in men / 亚洲男科学杂志(英文版)

This study was designed to characterize morphometric sperm subpopulations in normozoospermic men by using different statistical methods and examining their suitability to classify correctly different sperm nuclear morphologies present in human ejaculates. Ejaculates from 21 normozoospermic men were collected for the study. After semen collection and analysis, samples were prepared for morphometric determination. At least 200 spermatozoa per sample were assessed for sperm morphometry by computer-assisted sperm morphometry analysis (CASA-Morph) using fluorescence. Clustering and discriminant procedures were performed to identify sperm subpopulations from the morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three morphometric subpopulations (large-round 30.4%, small-round 46.6%, and large-elongated 22.9%). In the second analysis, using discriminant methods, the classification was made independently of size and shape. Three morphological categories according to nuclear size (small 13.07 μm2) and four categories were defined on 400 canonical cells (100 × 4) from 10 men according to sperm nuclear shape (oval, pyriform, round, and elongated). Thereafter, the resulting classification functions were used to categorize 4200 spermatozoa from 21 men. Differences in the class distribution were observed among men from both clustering and discriminant procedures. It was concluded that the combination of CASA-Morph fluorescence-based technology with multivariate cluster or discriminant analyses provides new information on the description of different morphometric sperm subpopulations in normal individuals, and that important variations in the distribution of morphometric sperm subpopulations may exist between men, with possible functional implications.

Characterization of Senescence of Culture-expanded Human Adipose-derived Mesenchymal Stem Cells

BACKGROUND AND OBJECTIVES: Adipose-derived mesenchymal stem cells (ADSCs) are promising candidates in regenerative medicine. The need for in vitro propagation to obtain therapeutic quantities of the cells imposes a risk of impaired functionality due to cellular senescence. The aim of the study was to analyze in vitro senescence of previously cryopreserved human ADSCs subjected to serial passages in cell culture. METHODS AND RESULTS: ADSC cultures from 8 donors were cultivated until proliferation arrest was reached. A gradual decline of ADSC fitness was observed by altered cell morphology, loss of proliferative, clonogenic and differentiation abilities and increased β-galactosidase expression all of which occurred in a donor-specific manner. Relative telomere length (RTL) analysis revealed that only three tested cultures encountered replicative senescence. The presence of two ADSC subsets with significantly different RTL and cell size was discovered. The heterogeneity of ADSC cultures was supported by the intermittent nature of aging seen in tested samples. CONCLUSION: We conclude that the onset of in vitro senescence of ADSCs is a strongly donor-specific process which is complicated by the intricate dynamics of cell subsets present in ADSC population. This complexity needs to be carefully considered when elaborating protocols for personalized cellular therapy.

Ejercicio y redes fisiológicas y bioquímicas en la hipertensión / Exercise training and functional networks of physiological and biochemical variables in hipertensión / Treinamento físico e redes funcionais de variáveis bioquímicas fisiológicas em hipertensão

El objetivo de este estudio fue evaluar el efecto del ejercicio sobre varios biomarcadores. Por primera vez se investigaron los cambios causados por el ejercicio en algunas variables bioquímicas y fisiológicas utilizando un análisis estructural. Los pacientes hipertensos fueron divididos en grupo de ejercicio (EXG) y grupo de no ejercicio (NoEXG). Se determinaron el perfil lipídico, subpoblacionesde HDL, apoAI, MMP-2 y MMP-9, al inicio del estudio y después de 8 semanas de intervención. Los datos muestran una variación significativa relacionada con el ejercicio en HDL3 (p=0,0236) y la presión sistólica (p=0,0448). También muestran que antes de hacer ejercicio las variables bioquímicas y fisiológicas estaban compartimentalizadas e independientes, mientras que después del ejercicio se establecieron relaciones entre ellas. Este estudio demuestra que un programa de entrenamiento regula la presión arterial y las HDL3, HDL2 y MMP-2. Pero la principal conclusión es que el ejercicio físico es eficaz en la generación o restauración de la red de relaciones entre las variables fisiológicas y las variables bioquímicas tales como presión sanguínea MMP-2, MMP-9, apoA-I, HDL2 y HDL3.

The aim of this study was to evaluate the effect of exercise on several biomarkers. For the first time, the changes caused by exercise training in physiological relationships were investigated using structural analysis. Hypertensive patients were divided into exercise group (EXG) and not exercise group (NEXG)). Lipid profile, HDL subpopulations, apoAI, MMP-2 and MMP-9 were assessed at baseline and after 8 weeks of intervention. Data show a significant exercise-related variation in HDL3 (p=0.0236) and SBP (p=0.0448). The results showed that, before exercise, biochemical and physiological variables were compartmentalized and independent; while after exercise these variables were able to establish relationships. This study demonstrates that an exercise training program significantly reduces systemic BP and HDL3 and increases HDL2 and MMP-2. But the main finding is that physical exercise is effective in generating or restoring a network of relationships between physiological variables loke BP and biochemical variables like MMP-2, MMP9, apoA-I, HDL2 and HDL3.

O objetivo deste estudo foi avaliar o efeito do exercício físico sobre vários biomarcadores. Pela primeira vez, as alterações causadas pelo exercício físico em algumas variáveis bioquímicas e fisiológicas foram investigadas através de uma análise estrutural. Os pacientes hipertensosforam divididos em grupo de exercício (EXG) e grupo de não-exercício (NãoEXG). Perfil lipídico, subpopulações de HDL, apoAI, MMP-2 e MMP-9, foram determinados no início do estudo e após oito (8) semanasde intervenção. Os dados mostram uma variação significativa associada ao exercício em HDL3 (p=0,0236) e pressão sistólica (p=0,0448). Mostram, também, que antes de fazer exercício, variáveis bioquímicas e fisiológicas estavam compartimentadas e independentes, ao passo que após o exercício foram estabelecidas relações entre elas. Este estudo demonstra que um programa de treinamento regula a pressão arterial e HDL3, HDL2 e MMP-2. Mas a principal conclusão é que o exercício físico é eficaz gerando ou restaurando a rede de relações entre variáveis fisiológicas e bioquímicas, tais como a pressão sanguínea MMP-2, MMP-9, apo AI, HDL2 e HDL3.