Flowcytometric immunophenotyping of peripheral-blood leukocytes in relation to immunopathology and cellular proliferation of pleural mesothelioma

Immunomodulation of malignant mesothelioma (MM) is variable considering host immune mechanisms. This study has investigated the immunophenotyping of peripheral-blood leukocytes of patients with mesothelioma in relation to different histopathologic types and proliferative potentiality. The study was achieved on 18 patients presented by massive effusion accompanying MM. Circulating CD4+ and CD8+ T cells were counted using two-colour flowcytometry. Pleural effusion cell block preparation and pleural biopsies were processed for immunohistochemical analysis of intratumoral CD8+ lymphocytes, cytokeratin and vimentin antigens expression, and modified one step silver stain to identify the Argyrophilic nucleolar organizer regions (AgNors) for evaluation of the proliferative potentiality of the tumor. Blood samples of five normal individuals were considered as controls. Histopathologic examination demonstrated epithelial variant in 12 cases, mixed (biphasic) variant in 4 cases and 2 cases were sarcomatoid. Flowcytometric analysis exhibited a significant increase in the mean percentage of circulating CD4+ T cell count in mesothelioma (56.7% +/- 6.99) versus control (43% +/- 1.58), p < 0.001. In contrast, cytotoxic CD8+ cells were significantly reduced (p < 0.001) in comparison to the control (25.2% +/- 2.96 versus 32% +/- 4.6 respectively). CD4/CD8 ratio was significantly increased versus control (2.06 +/- 0.57 versus 1.3 +/- 0.02 p < 0.05). Intratumoral CD8+ lymphocytes were depleted in mesothelioma cases. There was significant reduction of circulating cytotoxic T lymphocytes (CTL) in relation to mixed and sarcomatoid variants versus epithelial differentiation (mean = 25.25 +/- 4.86, 22 +/- 1.41 and 29.5 +/- 9.9 respectively with a p value < 0.05). Reduction of cytotoxic T cells was also associated with increased expression of vimentin and increased proliferative activity which has been identified by increased AgNors in mixed and sarcomatoid differentiation. In conclusion, mesothelioma might be associated with modulation of the tumoricidal effect of cytotoxic T lymphocytes in relation to tumor differentiation and its proliferative potentiality. Immunophenotyping analysis of leukocytes may reflect the competence of immune system against malignancy and act as an additive prognostic parameter for mesothelioma progression and probably expecting response to oncotherapy protocols.     

Human natural killer T cells (NKT), NK and T cells in pulmonary tuberculosis: potential indicators for disease activity and prognosis

One third of the world's population is infected with Mycobacterium tuberculosis (MTB). However, active disease can develop only in a small percentage, when the immunity is weakened. The acquired immune response to MTB is primarily mediated by T cells. Natural killer (NK) cells play a central role in innate immunity to microbial pathogens. Human NKT cells have characteristics of both T and NK cells and also exhibit antimycobacterial activity. This work aimed to enumerate T, NK and NKT cells in active pulmonary TB compared with healthy controls and to study the correlation between these cells with different factors affecting prognosis of pulmonary TB as disease severity, complications or associated diseases, antitubrculosis chemotherapy, and age & gender. Of the 22 active tuberculosis patients examined, 17 were recent cases and 5 recurrent. Healthy controls were divided into 14 individuals with detectable reaction to purified protein derivative (PPD+) and 14 individuals without detectable reaction to PPD-. The percentages of T, NK and NKT cells in erythrocyte-lysed whole blood samples were determined using flowcytometry. The percentage of NKT cells was significantly higher among the recently diagnosed MTB cases as compared with both PPD+ (P < 0.01) and PPD- (P < 0.01) healthy controls, while no significant difference could be found in the percentages of T or NK cells among these groups. However, comparing recurrent cases with recently diagnosed cases showed a significant difference only in the percentage of T cells (P < 0.01). There was also a significant difference in the percentage of T cells according to severity of disease (P < 0.01) and in the association of diabetes mellitus (P < 0.01). Age, gender and treatment with antituberculosis chemotherapy had no effect on the percentages of T, NK or NKT cells. It is concluded that T and NKT cells play an important role in immunity against TB. In active pulmonary tuberculosis, increased T cell count points to severity of the disease, while their reduced count predicts bad prognosis. Human NKT cell count is a marker of disease activity. Enumeration of these cells in peripheral blood can be used as a non-invasive prognostic indicator for patients with active pulmonary TB.     

Possible role of angiotensin-converting enzyme polymorphism on progression of hepatic fibrosis in chronic hepatitis C virus infection

Many functional polymorphisms in the rennin-angiotensin system (RAS) have been described; these polymorphisms have been postulated to contribute to fibrosis in several diseases. Our aim was to study the frequency of ACE I/D polymorphism in chronic hepatitis C virus (HCV) infection and its association with liver fibrosis and response to treatment. This study included 90 patients with chronic hepatitis C. All patients received antiviral therapy in the form of pegylated interferon and ribavirin. Patients were grouped according to the stage of liver fibrosis by biopsy into: group 1 (fibrosis: 0 or 1); group 2 (fibrosis: 2 or 3) and group 3 (fibrosis: 4 or 5). The study included also 170 healthy subjects, as a control group. Polymerase chain reaction was carried out to detect the different ACE genotypes. The D/D genotype was significantly more prevalent among HCV patients compared to controls (65.6% vs 48.2%, P = 0.006). Degree of necroinflammation was significantly higher among patients with I/I genotype when compared to patients with D/D genotype (P = 0.04). No significant difference in the distribution of the ACE I/D genotypes between the fibrosis groups and between responders and non responders to interferon therapy. The D/D genotype may increase the susceptibility to infection with hepatitis C.     

Analysis of apoptosis and a Th1/Th2 phenotype in HIV-infected patients

Lymphocytes from HIV patients, unlike those from normal HIV-negative subjects, underwent apoptosis upon in vitro culture. We found that the percentage of lymphocytes undergoing apoptosis was significantly higher (P = 0.005) in patients with low CD4 cell counts (< 200 CD4 cells/μl) (60%) than in patients at earlier stage (> 500 CD4 cells/μl) (35%). Serum IgE levels increased in two of six patients at last stage and in two of five patients at earlier stage. Spontaneous production of both IL-2 and IL-10, by peripheral blood mononuclear cells (PBMC) after 48 h in culture, was greater in HIV-infected subjects and increased with disease progression. IFN-γ production was greater in HIV-infected subjects but there was no evident change with disease progression. IL-4 production was barely detectable or not detected in both HIV-infected and HIV-negative individuals. These results indicate that spontaneous apoptosis is associated with advanced disease. However, there was no evidence of in vivo switch from the Th1 to Th2 phenotype in HIV-infected patients.     

Health benefits of cyanidin-3-glucoside as a potent modulator of Nrf2-mediated oxidative stress

Inflammopharmacology - Berries are natural sources of anthocyanins, especially cyanidin-3-glucoside (C3G), and exhibit significant antioxidant, antidiabetic, anti-inflammatory, and cytoprotective...     

The science of Durban,AIDS 2016

Introduction : The science presented at the 21st International AIDS Conference in Durban, South Africa, in July 2016, addressed the state of the field across basic, clinical, prevention, law and policy and implementation science. Methods and Results : The AIDS response has seen remarkable achievements in scientific advances, in translation of those advances into prevention, treatment and care for affected individuals and communities, and in large scale implementation – reaching 18 million people with antiviral therapy by mid‐year 2016. Yet incident HIV infections in adults remain stubbornly stable and are increasing in some regions and among adolescents and adults in some key populations, challenging current science, policy and programming. There have been important advances in both preventive vaccines and in cure research, but both areas require ongoing investment and innovation. Clinical research has flourished with new agents, regimens, delivery modes and diagnostics but has been challenged by aging and increasingly complex patient populations, long‐term adherence challenges, co‐infections and co‐morbidities, and unresolved issues in TB management and epidemic control. It is an extraordinary period of innovation in prevention, yet the promise of new tools and combination approaches have yet to deliver epidemic HIV control. Conclusions : Proven interventions, most notably pre‐exposure prophylaxis, PrEP, have been limited in rollout and impact. Treatment as prevention has the promise to improve clinical outcomes but remains uncertain as a prevention tool to reduce population‐level HIV incidence. The improvement of legal, policy and human rights environments for those most at risk for HIV acquisition and most at risk for lack of access to essential services; sexual and gender minorities, sex workers of all genders, people who inject drugs, and prisoners and detainees remain among the greatest unmet needs in HIV/AIDS. Failure to do better for these individuals and communities could undermine the HIV response.     

Chemically modified carbon-based electrodes for the determination of paracetamol in drugs and biological samples

Paracetamol is a non-steroidal, anti-inflammatory drug widely used in pharmaceutical applications for its sturdy, antipyretic and analgesic action. However, an overdose of paracetamol can cause fulminant hepatic necrosis and other toxic effects. Thus, the development of advantageous analytical tools to detect and determine paracetamol is required. Due to simplicity, higher sensitivity and selectivity as well as costefficiency, electrochemical sensors were fully investigated in last decades. This review describes the advancements made in the development of electrochemical sensors for the paracetamol detection and quantification in pharmaceutical and biological samples. The progress made in electrochemical sensors for the selective detection of paracetamol in the last 10 years was examined, with a special focus on highly innovative features introduced by nanotechnology. As the literature is rather extensive, we tried to simplify this work by summarizing and grouping electrochemical sensors according to the by which manner their substrates were chemically modified and the analytical performances obtained.     

Hemato-immunologic impact of subchronic exposure to melamine and/or formaldehyde in mice

The aim of the present study was to explore the potential hematotoxic and immunotoxic effects of melamine (MA) in the absence and presence of formaldehyde (FA) in mice. Forty adult Swiss mice were equally allocated into four groups and daily treated with water, MA (50?mg/kg), FA (25?mg/kg), and MA?+?FA respectively via feeding needle for 60 consecutive days. Hematological status was evaluated using erythrogram and leukogram profiling. Innate immune functions were assessed by measuring white blood cells lysozyme and phagocytic activities. Serum immunoglobulin levels were evaluated as indicators of humoral immunity. In addition, histologic and immunohistochemical evaluations of splenic tissues were performed. The results indicated that either MA or FA treatment resulted in significant decreases in RBCs, Hb, MCHC, total WBC, lymphocyte, and basophile levels as well as in WBCs phagocytosis and lysozyme activity. In contrast, MCV, PCV%, and reticulocyte levels were significantly increased in these hosts. The total IgM level was significantly reduced in the MA-only-exposed mice but markedly increased in the FA-only-treated ones. A significant decrease in serum IgG levels was detected following either MA or FA treatment. The combined exposure to MA and FA, compared to levels of either toxicant alone, was revealed to evoke a significant improvement in Hb, PCV%, MCV, MCHC, neutrophil, eosinophil, total IgM level, and lysozyme activity; however these values did not reach that of the controls. Furthermore, compared to control mice, both MA-only- and FA-only-treated mice showed a strong distribution of CD4⁺ and CD8⁺ cells in their spleens, while a moderate presence of the former cells was obvious at their co-exposure. Taken together, these findings revealed that exposure to MA or FA resulted in significant alterations in hemato-immune parameters at variable degrees while a co-exposure resulted in the mitigation of most effects of either toxicant alone.