Quantification and Comprehensive Analysis of Mesenchymal Stromal Cells in Bone Marrow Samples from Sickle Cell Disease Patients with Osteonecrosis.

The potential use of bone marrow mesenchymal stromal cells (BM-MSCs) for the treatment of osteonecrosis in sickle cell disease (SCD) patients is increasing. However, convenient BM-MSC quantification and functional property assays are critical factors for cell-based therapies yet to be optimized. This study was designed to quantify the MSC population in bone marrow (BM) samples from SCD patients with osteonecrosis (SCD group) and patients with osteoarticular complications not related to SCD (NS group), using flow cytometry for CD271+CD45-/low cell phenotype and CFU-F assay. We also compared expanded BM-MSC osteogenic differentiation, migration, and cytokine secretion potential between these groups. The mean total cell number, CFU-F count, and CD271+CD45-/low cells in BM mononuclear concentrate were significantly higher in SCD than in NS patients. A significant correlation between CD271+CD45-/low cell number and CFU-F counts was found in SCD (r = 0.7483; p = 0.0070) and NS (r = 0.7167; p = 0.0370) BM concentrates. An age-related quantitative reduction of CFU-F counts and CD271+CD45-/low cell number was noted. Furthermore, no significant differences in the morphology, replicative capacity, expression of surface markers, multidifferentiation potential, and secretion of cytokines were found in expanded BM-MSCs from SCD and NS groups after in vitro culturing. Collectively, this work provides important data for the suitable measurement and expansion of BM-MSC in support to advanced cell-based therapies for SCD patients with osteonecrosis.

Specific Cytostatic and Cytotoxic Effect of Dihydrochelerythrine in Glioblastoma Cells: Role of NF-κB/ß-catenin and STAT3/IL-6 Pathways.

BACKGROUND: A glioblastoma is a primary CNS tumor that is more aggressive and lethal than other brain tumors. Its location, rapid proliferation, invasive growth, angiogenesis and immunosuppression are the main factors that limit its treatment, making it a major challenge to neuro-oncology. OBJECTIVE: This study investigated the in vitro effects of the alkaloid dihydrochelerythrine (DHC), which is extracted from Zanthoxylum stelligerum, on the viability, proliferation, cell death and ß-catenin, NFκB, STAT3/pSTAT3 and interleukins roles. METHOD: In vitro experimental models of human (U251 and GL-15) and murine (C6) glioblastoma cells were cultured in the presence of DHC at increasing concentrations for MTT assay and exclusion trypan blue dye to determine EC50. Afterward, C6 and U251 cells were treated with 100 µM DHC or DMSO 0.1% for cell cycle, annexin and expression of ß-catenin/NFκB/STAT3/pSTAT3 by flow cytometry or immunofluorescence. Interleukin quantification was made by Cytometric Bead Array. RESULTS: A significant decrease was observed in C6 and U251 cell viability in a time and dose-dependent manner. GL-15 cell viability decreased only when treated with 200 µM DHC. This maximum concentration affected neither astrocytes nor microglia viability. A cytostatic effect of DHC was observed in C6 and U251 cells after 48 h of 100 µM DHC treatment. After 72 h of DHC treatment, C6 presented 80% of annexin-V+ cells compared to 10% of annexin-V+ U251 cells. C6 cells demonstrated significant high levels of NFκ B and ß-catenin cytoplasmic fraction. Additionally, DHC treatment resulted in higher significant levels of IL-6 than did other interleukins and STAT3 up-regulation in U251 cells. CONCLUSION: These results demonstrate that DHC acts as a chemosensitizing agent selective for glioma cells not affecting non-tumor cells. Considering tumor heterogeneity, DHC demonstrated an anti-cancer potential to activate different cell death pathways. DHC demonstrated could be used for chemotherapy and immunotherapy applications in glioblastomas in the future.

Lymphocyte subset reference intervals in blood donors from northeastern Brazil.

UNLABELLED: The reference intervals for leukocytes and lymphocytes currently used by most clinical laboratories present limitations as they are primarily derived from individuals of North American and European origin. The objective this study was to determine reference values for peripheral blood B lymphocytes, T lymphocyte subsets (CD4+, CD8+, naïve, memory, regulatory, TCRαß and TCRγδ+) and NK cells from blood donors in Salvador-Bahia, Brazil. RESULTS: The proportion of included male subjects was 73.7% and the median ages of males (34) and females (35) were found to be similar. Absolute counts total lymphocytes subsets to both gender was 1,956 (1,060-4,186) cells and relative values 34%. The T CD4+ and T CD8+ lymphocytes relative values was 51% (20-62) and 24% (9-28), respectively. The most statistically significant finding observed was a higher percentage of B lymphocytes (p=0.03) in females. Commonly cited subset reference intervals were found to be consistent with values in several populations from different geographic areas.

Hepatitis B virus prevalence and vaccination response in health care workers and students at the Federal University of Bahia, Brazil.

UNLABELLED: BACKGROUND AND RATIONALE FOR THE STUDY: Hepatitis B (HB) is one of the most prevalent occupational infections in health attendance environments. According to the Brazil Ministry of Health, health professionals must be vaccinated against the hepatitis B virus (HBV) and provide laboratory proof of immunization. AIMS: To evaluate the seroprevalence of HBV infection and to analyze the response to vaccine by measuring serum antibodies against HBV surface antigen (anti-HBs) levels in a sample of students and health professionals at the Federal University of Bahia. RESULTS: As part of this cross-sectional study, a campaign against occupational HB was launched in 2007 and vaccination and blood samples were collected for analysis of the following serological markers: HBV surface antigen (HBsAg) and anti-HBs (measured by enzyme-linked immunoassay) and total antibodies against HBV core antigen (anti-HBc). The study sample comprised 766 people. Global seropositivity for HBV was 1.7%: 0.5% in the students and 8.8% in the professionals. In a group of volunteers, a serological profile compatible with postvaccine immunity was shown by 95% of volunteers with proof of vaccination and by 81.8% of volunteers without proof of vaccination. CONCLUSIONS: In conclusion, this study shows that it is important to promote vaccination campaigns and improve knowledge and awareness about HB among health care workers and higher education students.

Ancestry informative markers and complete blood count parameters in Brazilian blood donors / Marcadores informativos de ancestralidade e parâmetros no hemograma de doadores de sangue brasileiros

A complete blood count is very useful in clinical diagnoses when reference ranges are well established for the population. Complete blood counts and allele frequencies of Ancestry Informative Markers (AIMs) were analyzed in Brazilians with the aim of characterizing the hematological values of an admixed population. Positive associations were observed between gender and neutrophils, monocytes, eosinophils, erythrocytes, hemoglobin, hematocrit, MCV, MCHC and platelet counts. No significant differences were found for age, alcohol consumption, educational status, ethnicity, smoking in respect to the complete blood count values. In general, men had higher red blood cell values, while women had higher values for white blood cells and platelets. The study of the population was highly heterogeneous with mean proportions (± SE) of African, European and Amerindian ancestry being 49.0 ± 3.0 percent, 44.0 ± 9.0 percent and 7.0 ± 9.0 percent, respectively. Amerindian ancestry showed limited contribution to the makeup of the population, but estimated ancestral proportions were statistically significant (r = 0.9838; P<0.001). These hematologic values are similar to Afro-Americans, another admixed population.

O hemograma é muito útil no diagnóstico quando o intervalo de referência é adequadamente estabelecido para população. Com o objetivo de verificar os valores hematológicos em população heterogênea foi analisado o hemograma e frequências alélica de marcadores informativos de ancestralidade de brasileiros. Foi observada associação positiva entre sexo e os valores de neutrófilos, monócitos, eosinófilos, eritrócitos, hemoglobina, hematócrito, MCV, MCHC e plaquetas (IC 95 por cento; P<0,05). E não houve diferenças entre idade, consumo de álcool, nível educacional, etnia, tabagismo e os valores do hemograma (IC 95 por cento; P>0,05). Os homens apresentaram valores maiores no eritrograma, enquanto no leucograma e plaquetograma foram as mulheres. Foi observado também que a população é altamente heterogênea e as médias proporcionais (±DP) de ancestralidade Africana, Europeia e Ameríndia estimada foram: 49,0 ± 3,0 por cento, 44,0 ± 9,0 por cento e 7,0 ± 9,0 por cento, respectivamente. A contribuição ancestral ameríndia se demonstrou pequena, mas a estimativa de proporções ancestrais foi estatisticamente significante (r = 0,9838; P<0,001). Os valores hematológicos aqui descritos são parecidos com os descritos em negros americanos, outra população heterogênea.

Serum antibody levels against Porphyromonas gingivalis extract and its chromatographic fraction in chronic and aggressive periodontitis.

UNLABELLED: Periodontitis is an inflammatory oral disease caused by multifactorial intrinsic and extrinsic agents, including Gram-negative bacteria such as Porphyromonas gingivalis. OBJECTIVE: To evaluate if there is difference in the serum levels of IgA, IgG and IgG subclasses reactive to Porphyromonas gingivalis in subjects with different periodontal conditions. METHODS: The study included 89 patients divided into four groups: 29 subjects with moderate or severe chronic periodontitis (CP), 12 with aggressive periodontitis (AP), 22 with gingivitis or mild periodontitis (GP), and 26 healthy controls (HC). Humoral response was assayed by enzyme-linked immunosorbent assay (ELISA) to verify serum levels of IgG, IgG1, IgG2, IgG3, IgG4 and IgA serum levels reacting to crude P. gingivalis ATCC 33277 sonicate extract and fraction IV, an enrichment of the immunoreactive bands of the crude extract obtained by chromatography. RESULTS: IgA, IgG (p < 0.01), IgG2, IgG3 and IgG4 serum level reactions to fraction IV were higher in the CP group compared with the healthy control. The CP group had higher levels of IgG and IgG4 to both antigens than the GP group, and higher levels of IgG and IgG4 to sonicate extract than the AP group. There were statistically significant differences in serum levels of IgG to both antigens (p < 0.01), IgG2 to fraction IV (p < 0.01), IgG3 to fraction IV (p < 0.05) and IgG4 to both antigens (p < 0.05) between AP and HC groups. IgG1 titers to sonicate extract were significantly higher (p < 0.05) in the GP group in comparison to the AP group. CONCLUSIONS: The findings of this study suggest that there are differences in the serum levels of IgA, IgG and IgG subclasses in patients with different periodontal conditions.

Soroprevalência de infecção pelo vírus da hepatite B e resposta de anticorpos anti-HBS à vacinação em cirurgiões-dentistas da grande Salvador-Bahia / Hepatitis B virus seroprevalence and anti-HBS serum response in dental surgeons in salvador-Bahia

De acordo com o Ministério da Saúde do Brasil, a hepatite B destaca-se como uma doença de transmissão ocupacional na prática odontológica, causando interrupções precoces na vida profissional dos dentistas. Portanto é fundamental recorrer-se às imunizações antes do início da vida profissional. Os objetivos deste trabalho foram avaliar a situação vacinal e soroprevalência para hepatite B em uma amostra de 120 cirurgiões-dentistas (CDs) de Salvador - Bahia. Os dados clínico-epidemiológicos foram obtidos por questionário individual, auto-aplicativo. Os marcadores sorológicos anti-HBs, anti-HBc total e AgHBs foram dosados por ensaio imunoenzimático quimioluminescente automatizado. Os resultados mostraram que 5,83por cento apresentaram positividade para o anti-HBc total, porém nenhuma das amostras mostrou positividade no teste de identificação do AgHBs. Observou-se que a maioria dos indivíduos estudados receberam quatro doses da vacina (53,4por cento). Respeitaram os intervalos entre as doses 72,1por cento dos indivíduos e 21,7por cento apresentaram níveis séricos de anti-HBs inferiores a 10mUI/ml s. O índice de acidentes com material pérfuro-cortante foi elevado, muito embora apenas um diminuto número de profissionais procurou tomar medidas preventivas recomendadas pós-acidente. O estudo sugere a realização de campanha de conscientização a respeito da importância da avaliação sorológica pós-vacinal, assim como da profilaxia pós-exposição.

Existe correlação entre níveis séricos de anticorpos reativos a porphyromonas gingivalis e a condição periodontal clínica? / Is there any correlation between serum antibody levels reactive to porphyromonas gingivalis and clinical periodontal status?

Porphyromonasgingivalis has been shown to be shown to be one of the most important pathogen in chronic periodontitis development. The aim of this study was to evaluate the correlation among IgA, IgG and IgC and IgG isotype serum levels reactive Porphyromonas gingivalis ATTCC33277 and the clinical periodontal status. Twenty nine periodontitis patients were evaluated according to clinical parameters: probing depth, bleeding on probing, and clinical attachment loss measurements. Humoral immune response was assayed by enzyme linked immunosorbent assay (ELISA). There was no correlation between bleeding on probing (criterion 1) and the evaluated antibodies. Significant positive correlations were found between IgA levels and percentual of sites with clinical attachment loss (CAL) >3mm (criterion 2), percentual of sites with CAL > 5 mm (criterion 3), and percentual of sites with CAL > 3mm associated to probing depth > 4mm and bleeding on probing at the same site (criterion 4) (r=0,311; r=0,336; r=0,358, respectively; p<0,05). There were significant positive correlations between IgG serum levels and criteria 3 and 4 (r=0,400 and r=0,372, respectively; p<0,05), and between IgG1 and criteria 2, 3 and 4 (r=0,333, r=0,323 and r=0,340, respectively; p<0,05). The findings indicate that the antibody response to Porphyromonas gingivalis depends on the periodontal status, determined by clinical parameters

Humoral immune response to antigens of Porphyromonas gingivalis ATCC 33277 in chronic periodontitis

Introduction: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. Objective: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. Methods: Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP), 10 patients with gingivitis (G) and 21 periodontally healthy individuals (H) were measured by ELISA and Western immunoblotting. Results: In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p<0.001; Dunnet t3 p= 0.001 and Dunnet t3 p= 0.0001). There was no statistically significant difference between G and HP reactivity (Dunnett t3 p=0.617). Among individuals with chronic periodontitis, the IgG-anti-P. gingivalis serum levels were positively correlated with percentage of clinical attachment level =5mm (r s = + 0.375, p<0.05) and a negative correlation was found between IgG-anti-P. gingivalis levels and percentage of probing pocket depth 0-3mm (r s = - 0. 411, p< 0.05). The analysis of sera immunoreactivity profiles to sonicate antigen by Western blotting showed differences between the sera of CP, G and H group individuals. The serum from CP frequently reacted with high molecular weight (103 kDa, 86 kDa, 72 kDa, 60 kDa, 58 kDa, 52 kDa) protein fractions. Conclusions: Serum levels of IgG anti-P. gingivalis distinguished individuals with chronic periodontitis, gingivitis and healthy periodontium. There was a correlation between clinical parameters and serum IgG levels against P. gingivalis. There was a difference in the recognition profile of protein fractions among the studied groups and some bands were more specific.

Humoral immune response to antigens of Porphyromonas gingivalis ATCC 33277 in chronic periodontitis.

INTRODUCTION: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. OBJECTIVE: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. METHODS: Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP), 10 patients with gingivitis (G) and 21 periodontally healthy individuals (H) were measured by ELISA and Western immunoblotting. RESULTS: In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p<0.001; Dunnet t3 p= 0.001 and Dunnet t3 p= 0.0001). There was no statistically significant difference between G and HP reactivity (Dunnett t3 p=0.617). Among individuals with chronic periodontitis, the IgG-anti-P. gingivalis serum levels were positively correlated with percentage of clinical attachment level =5mm (r s = + 0.375, p<0.05) and a negative correlation was found between IgG-anti-P. gingivalis levels and percentage of probing pocket depth 0-3mm (r s = - 0. 411, p< 0.05). The analysis of sera immunoreactivity profiles to sonicate antigen by Western blotting showed differences between the sera of CP, G and H group individuals. The serum from CP frequently reacted with high molecular weight (103 kDa, 86 kDa, 72 kDa, 60 kDa, 58 kDa, 52 kDa) protein fractions. CONCLUSIONS: Serum levels of IgG anti-P. gingivalis distinguished individuals with chronic periodontitis, gingivitis and healthy periodontium. There was a correlation between clinical parameters and serum IgG levels against P. gingivalis. There was a difference in the recognition profile of protein fractions among the studied groups and some bands were more specific.