Tunicamycin-induced Endoplasmic Reticulum Stress Upregulates the Expression of Pentraxin 3 in Human Retinal Pigment Epithelial Cells

PURPOSE: To investigate the production of long pentraxin 3 (PTX3) in response to tunicamycin-induced endoplasmic reticulum (ER) stress and its role in ER stress-associated cell death, PTX3 expression was evaluated in the human retinal pigment epithelial cell line, ARPE-19. METHODS: PTX3 production in ARPE-19 cells was analyzed in the absence or presence of tunicamycin treatment by enzyme-linked immunosorbent assay. PTX3 protein and mRNA levels were estimated using western blot analysis and real-time reverse transcription-polymerase chain reaction, respectively. Protein and mRNA levels of CCAAT-enhancer-binding protein homologous protein (CHOP) and ARPE-19 cell viability were measured in the presence of tunicamycin-induced ER stress in control or PTX3 small hairpin RNA (shRNA)-transfected ARPE-19 cells. RESULTS: The protein and mRNA levels of PTX3 were found to be significantly increased by tunicamycin treatment. PTX3 production was significantly decreased in inositol-requiring enzyme 1α shRNA-transfected ARPE-19 cells compared to control shRNA-transfected cells. Furthermore, pretreatment with the NF-κB inhibitor abolished tunicamycin-induced PTX3 production. Decreased cell viability and prolonged protein and mRNA expression of CHOP were observed under tunicamycin-induced ER stress in PTX3 shRNA transfected ARPE-19 cells. CONCLUSIONS: These results suggest that PTX3 production increased in the presence of tunicamycin-induced ER stress. Therefore, PTX3 could be an important protector of ER stress-induced cell death in human retinal pigment epithelial cells. Inositol-requiring enzyme 1α and the NF-κB signaling pathway may serve as potential targets for regulation of PTX3 expression in the retina. Therefore, their role in PTX3 expression needs to be further investigated.

C - reactive protein levels in cerebral infaarctioninpatients on hemodialysis

Serum C-reactive protein [CRP] concentration issensitive marker of underlying systemic inflammation. Patientsunder continuous hemodialysis have an activated inflammatoryresponse, evidenced by increased serum CRP levels especiallyin patients with cerebral stroke. The study was performed toevaluate serum levels of CRP as inflammatory markers inpatients with stroke under continuous hemodialysis. The study was included 33 patients with chronicrenal failure under continuous hemodialysis divided into twogroups. Group I included 23 patients with acute stroke. Group II [control group] included 10 patients without stroke. All patientswere subjected to complete history and clinical examinationwith special emphasis to history of co-morbid conditions,hypertension, diabetes mellitus and cardiovascular disease.Serum creatinine, urea, CBC, albumin, lipid profile, calcium,fasting and post prandial blood sugar, sodium, potassium, serumuric acid, phosphorus, C-reactive protein and other investigationwere evaluated. In addition axial CT or MRI was performed atadmission and after 72 hours. The following parameterswere significantly higher in groupe I when compared to group II;ages [t=3.5. p< 0.01], CRP [t=7.1, p<0.001], serum creatinine [tett, p<0.0] and blood urea [t=4.3, p<0.01], while serumlevels of Hb% and serum calcium were significantly lower ingroup I when compared to group II [t=3.1, p<0.01 and t=2.2,p<0.05 respectively]. On the hand no significant differences inthe other studied parameters between two groups. Serum levelsof CRP were positively correlated with INR [r=520, p<0.05] and negatively correlated with serum calcium [r=0.580, p<0.05] and serum albumin [r=-540, p<0.05]. On the other hand nosignificant correlations were found between CRP and otherstudied parameters. According to ROC curve between group Iand II in CRP the cutoff were greater than 12 with sensitivity,specificity, Positive Perdictive Value and Negative PredictiveValue were 100% by accuracy 100%. Elevatedserum CRP could be a predictor of cerebrovascular stroke indialysis patients. Therefore, regular determination of serumCRP may be helpful to detect early signs of tissue damage andasymptomatic inflammation in these patients

Patterns of co-association of C-reactive protein and nitric oxide in malaria in endemic areas of Iran

In addition to numerous immune factors, C-reactive protein (CRP) and nitric oxide (NO) are believed to be molecules of malaria immunopathology. The objective of this study was to detect CRP and NO inductions by agglutination latex test and Griess microassay respectively in both control and malaria groups from endemic areas of Iran, including Southeastern (SE) (Sistan & Balouchestan, Hormozgan, Kerman) and Northwestern (NW) provinces (Ardabil). The results indicated that CRP and NO are produced in all malaria endemic areas of Iran. In addition, more CRP and NO positive cases were observed amongst malaria patients in comparison with those in control group. A variable co-association of CRP/NO production were detected between control and malaria groups, which depended upon the malaria endemic areas and the type of plasmodia infection. The percentage of CRP/NO positive cases was observed to be lower in NW compare to SE region, which may be due to the different type of plasmodium in the NW (Plasmodium vivax) with SE area (P. vivax, Plasmodium falciparum, mixed infection). The fluctuations in CRP/NO induction may be consistent with genetic background of patients. Although, CRP/NO may play important role in malaria, their actual function and interaction in clinical forms of disease remains unclear.

Effect of metabolic control on the in vitro proliferation of peripheral blood mononuclear cells in type 1 and type 2 diabetic patients / Efeito do controle metabólico na proliferação in vitro de células mononucleares de sangue periférico de pacientes diabéticos tipo 1 e 2

CONTEXT AND OBJECTIVE: Diabetes mellitus is a clinical syndrome that frequently leads to the development of chronic complications and high susceptibility to infections. It is probably due to defective immunological defense, which may be related to metabolic control of the disease. The aim of this study was to evaluate the effect of metabolic control on immune-cell behavior in type 1 and type 2 diabetic patients. For this, the in vitro proliferation of peripheral blood mononuclear cells (PBMC) was analyzed in patients with inadequate and adequate metabolic control. DESIGN AND SETTING: Experimental/laboratory study at a university hospital. METHODS: Eleven type 1 and thirteen type 2 diabetic patients were studied, together with 21 healthy individuals divided in two groups (11/10), who were matched by sex and age with those diabetic patients. PBMC cultures stimulated with concanavalin-A (Con-A) were used to measure ³H-thymidine incorporation after 72 hours of cell culturing. For patients with inadequate metabolic control, culturing was performed on the first day of patient hospitalization and again after intensive treatment to achieve adequate control. RESULTS: The proliferation index for Con-A-stimulated cultures from type 1 diabetic patients was significantly greater than that for cultures from healthy individuals and type 2 diabetic patients, independent of metabolic control. A negative correlation between the proliferation cell index and body mass index and serum C-reactive protein levels was also observed. CONCLUSION: The increase in the proliferation capacity of type 1 diabetic T lymphocytes was probably not caused by hyperglycemia and/or insulinopenia related to inadequate metabolic control.

CONTEXTO E OBJETIVO: Diabetes mellitus é uma síndrome clínica que freqüentemente leva ao desenvolvimento de complicações crônicas, e também, alta susceptibilidade a infecções, provavelmente devido a um defeito na defesa imunológica, que pode ser relacionada ao controle metabólico da doença. Neste estudo, propomos avaliar o efeito do controle metabólico no comportamento imunocelular de pacientes diabéticos tipo 1 e 2 através da proliferação in vitro de células mononucleares de sangue periférico (PBMC) em pacientes com controle metabólico inadequado e adequado. TIPO DE ESTUDO E LOCAL: Estudo experimental e laboratorial, realizado em hospital universitário. MÉTODOS: Estudamos 11 diabéticos do tipo 1 e 13 do tipo 2 além de 21 controles saudáveis, divididos em dois grupos (11/10), pareados para sexo e idade aos diabéticos tipo 1 e 2. Usamos culturas de PBMCs estimuladas com concanavalina (Con-A) para medir a incorporação de ³H-timidina após 72 horas de cultura de células. As culturas foram realizadas no primeiro dia de internação para os pacientes em controle metabólico inadequado, e repetidas após o controle metabólico adequado. RESULTADOS: O índice de estimulação das culturas estimuladas com Con-A nos diabéticos tipo 1 foi significantemente maior que na cultura de indivíduos saudáveis e diabéticos tipo 2, independentemente do controle metabólico. Além disso, foi observada uma correlação negativa entre o índice de proliferação e o índice de massa corporal e níveis de proteína-C. CONCLUSÃO: O aumento na capacidade de proliferação de linfócitos de diabéticos tipo 1 não é causado por hiperglicemia e/ou insulinopenia relacionada a controle inadequado.

Evaluation of C-reactive protein and other inflammatory markers on acute coronary syndromes

The aim of the study is to evaluate the role of C-reactive protein [CRP], ESR, total WBC and fibrinogen in acute myocardial infarction [AMI] and in unstable angina [UA], the two very common manifestations of acute coronary syndromes [ACS]. Subjects and The present study included 100 ACS patients with age ranging from 42-60 years [mean 53 +/- 6 years], of which 35 were presented with UA a n d 65 with AMI. The study also included 25 normal subjects, age and sex matched as controls. CRP levels were elevated in both A M I [40.8 +/- 15.4mg/l] as well as in UA [32.9 +/- 17.7mg/l] patients when compared to that of the normal c o n t rols [12.6 +/- 2.8mg/l]. But the value of fibrinogen was found to be elevated significantly only in AMI patients [381.9 +/- 32.1mg%]. Other markers such as WBC and ESR were found to be elevated in both AMI and UA patients. Elevation of CRP levels can be used as adjuncts in the diagnosis of ACS

The Effects of Antidepressant Treatment on Serum Cytokines and Nutritional Status in Hemodialysis Patients

The aim of this study was to investigate the effects of antidepressant treatment on serum cytokines and nutritional status in hemodialysis patients. Twenty-eight hemodialysis patients with a depressed mood were given 20 mg of fluoxetine for 8 weeks. The degree of depressive symptoms, the serum levels of interleukin-1beta, interleukin- 2, interleukin-6, tumor necrosis factor-alpha, c-reactive protein, and markers of nutritional status were assessed at baseline and after treatment. The outcome was assessed in terms of response to treatment (>50% reduction in the score of the Hamilton depression rating scale). Antidepressant treatment decreased the serum level of interleukin- 1 in both response and nonresponse groups, and increased the serum level of interleukin-6 only in the response group. At baseline, the level of interleukin-6 in the response group was lower than in the nonresponse group. Antidepressant treatment also increased fat distribution significantly in the response group which might have slightly improved the nutritional status. This study suggests that antidepressant treatment improve depressive symptoms and may affect immunological functions and nutritional status in chronic hemodialysis patients with depression.

High Dose Therapy Followed by Autologous Peripheral Blood Stem Cell Transplantation as a First Line Treatment for Multiple Myeloma: a Korean Multicenter Study

We conducted a phase II multicenter trial to estimate the response and survival of patients with newly diagnosed multiple myeloma to high dose melphalan therapy followed by autologous peripheral blood stem cell transplantation. Eligible patients who had undergone induction with vincristine, adriamycin and dexamethasone (VAD) should have adequate cardiac, pulmonary and renal function (creatinine <2 mg/dL). Melphalan at 200 mg/m2 was used as a conditioning regimen. Eighty patients were enrolled from 13 centers. The median age of the patients was 53 yr (range; 20 to 68 yr). The initial stage was IA/IIA/IIB/IIIA/IIIB in 3/8/1/54/14 patients, respectively. Beta2-microglobulin, CRP and LDH were increased in 74, 42 and 34% of the patients examined. Cytogenetic data were available in 30 patients, and 6 patients showed numeric or structural abnormalities. Two therapy-related mortalities occurred from infection. Among the 78 evaluable patients, CR/PR/MR/NC/PD were achieved in 48/26/2/1/1patients, respectively. After a median follow-up of 30 months, the median overall and event-free survivals were 66 months (95% CI: 20-112) and 24 months (95% CI: 18-29), respectively. This study verifies the efficacy and feasibility of high dose melphalan therapy with autologous stem cell transplantation in newly diagnosed multiple myeloma.

Determinación de receptores de haptoglobina y proteína C reactiva en linfocitos de pacientes con patología autoinmune e infecciosa / Determination of haptoglobin and C reactive protein receptors in lymphocytes of patients with autoimmune and infectious diseases

En la actualidad, se desconoce el rol del aumento de proteína C recativa (PCR) y haptoglobina (Hp) en la respuesta de fase aguda. Algunos autores han postulado la posibilidad que estas proteínas intervengan en la regulación del sistema inmune. Nuestro estudio estuvo orientado a demostrar la presencia de receptores para Hp y PCR en linfocitos procedentes de niños sanos y niños con patología infecciosa y autoinmune. Para este efecto, se obtuvieron células mononucleares de 48 niños (24 sanos, 14 con infecciones demostradas y 10 con enfermedades autoinmune), se incubaron por 72 horas a 37 grados Celsius y 5 por ciento de CO2, con estímulo de fitohemaglutinina (PHA) y con diferentes concentraciones de PCR y Hp en el medio. Se separaron las subpoblaciones CD4 y CD8 mediante anticuerpos monoclonales unidos a partículas magnéticas y se analizó la presencia de receptores a distintos tiempos (0, 24, 48 y 72 horas) mediante una técnica de inmunofluorescencia indirecta

Elevation of C-reactive protein in serum of Channa punctatus as an indicator of water pollution.

Effect of some pollutants like heavy metals, non-metals and pesticides on the circulating level of C-reactive protein (CRP) which is an acute phase plasma protein was studied in a freshwater murrel C. punctatus. Fish was exposed to nonlethal doses of these xenobiotics which were apparently safe. But the level of CRP detected by sensitive single radial immunodiffusion (SRID) technique showed that within 12 hr of exposure the nonlethal doses of xenobiotics could initiate the acute phase response in terms of elevated CRP titre. Heavy metals caused the acute phase within 24 hr, nonmetals and Metacid-50 within 48 hr exposure. The carbamate compound, carbaryl demonstrated a biphasic response to CRP level which may be correlated with the reversible type of anticholinesterase property of this compound while Metacid-50 is an irreversible type of anticholinesterase agent. The assessment of the CRP level in the serum of fish may be utilized as a primary bioindicator of a contaminated environment toxic enough to mount an acute phase response.