[MEFV gene mutation spectrum in familial Mediterranean fever (FMF) : a single center study in the Aegean region of Turkey]. / MEFV-Mutationsspektrum bei familiärem Mittelmeerfieber (FMF) : Monozentrische Studie aus der türkischen Ägäisregion.

OBJECTIVES: Familial Mediterranean fever (FMF), an autosomal recessive autoinflammatory disorder, is characterized by recurrent, self-limiting fever and serositis which is frequently seen in Mediterranean populations. In this study, we retrospectively evaluated the MEFV gene mutation distribution of 883 citizens of the Aegean region with preliminary diagnosis of FMF who were referred to the Tepecik Research and Education Hospital's Tissue Typing and Molecular Diagnostic Laboratory (Izmir, Turkey) between 2006 and 2009. METHODS: The FMF Strip Assay® (ViennaLab Diagnostics, Vienna, Austria) was used to determine the 12 most common MEFV gene mutations in patients prediagnosed with FMF. FINDINGS: Allelic frequencies of the major mutations in the mutation positive groups, including M694V, E148Q, M680I(G>C), and V726A, accounted for 48.4, 16.5, 13.5, and 9.7%, respectively. CONCLUSION: The M694V mutation was found to be the most common mutation among FMF patients in the Aegean region, which is in accordance with mutation studies reported from other regions of the country and different ethnic populations. An English full-text version of this article is available at SpringerLink as supplemental.

Diverse genotypical features and impacts on clinical course and severity of cystic fibrosis: early childhood experience.

AIM: Very little is known about the relationship between genotype and phenotype of cystic fibrosis (CF) from the Turkish children. The aim of the study was to analyze the genotype and phenotype of 24 children with CF and to investigate the correlation between type of mutation in cystic fibrosis transmembrane conductance regulator (CFTR) protein gene and clinical manifestation of the disease. METHODS: Patients were evaluated retrospectively and prospectively. History, clinical findings, sweat test and mutation analysis were used for the definitive diagnosis of CF. Phenotypical features of 24 cases were evaluated according to clinical findings. We compared the clinical phenotype and age at diagnosis, genotypic features. A total of 36 mutations were analyzed by polymerase chain reaction (PCR) and reverse hybridization methods. Statistical analysis was done by using χ2, Fisher exact and Pearson correlation tests. RESULTS: The mean age of the cases that were admitted to our out-patient clinic was 5.3±4 years. The median age of diagnosis was three months. Parents were consanguineous in 37.5% of cases and loss of a sibling at one year of age was stated in a quarter. The most frequent symptom was recurrent diarrhoea (79.2%) and there was severe growth retardation in 12 (50%) and pseudo-Bartter (PB) syndrome in 11 of the cases. The incidence of PB was higher in cases that were diagnosed at one year of age. Out of 18 cases with mutation analysis, nine (50%) were positive for DF508 mutation, and four cases were homozygous out of nine cases. Two separate mutations were determined in two cases with severe clinical picture. The incidence of respiratory tract infection during the admission was lower in DF508 positive cases (P=0.016). There was no statistically significant relation between DF508 positivity and diarrhea, severe growth retardation and PB (P>0.05). The other mutations that were determined in our patients were rarely seen mutations such as 3120+1 G-A, R347P, 1677delTA, 2789+5G-A, 2183AA-G, and R1066C. CONCLUSION: DF508 mutation rates in our cases diagnosed in early childhood were higher than the rates reported previously in Turkish children. The definition of molecular defect in CFTR gene has an impact on verifying the diagnosis and decreasing morbidity and mortality. An adequately large sample size is needed to evaluate the mutation profiles and genotype-phenotype characteristics in our country.

The effects of interferon-beta on interleukin-10 in multiple sclerosis patients.

The incidence of the neuropathological lesions and the severity of the clinical symptoms in multiple sclerosis (MS) are correlated with the amount of the transferred autoreactive T cells. The balance between the T helper 1 (Th1) and T helper 2 (Th2) cytokine phenotypes may affect the activity of the disease in MS patients. Interleukin-10 (IL-10) is a cytokine secreted by Th2 cells. Thus, it has been thought that inducing IL-10 may have therapeutic effects in the treatment of MS patients. In this study, in order determine whether different types of prophylaxis change the secretion of IL-10, we measured the levels of IL-10 in relapsing-remitting type multiple sclerosis (RRMS) patients receiving interferon-beta 1b (IFN-beta 1b) or azathioprine (AZA). Our study consisted of RRMS patients (n=45) and healthy subjects (n=15) as control group. Patients were categorized into three groups as those receiving either IFN-beta 1b or AZA and those not receiving prophylaxis. Each group was compared with the control group. Serum IL-10 levels were determined using ELISA method. IL-10 levels of those receiving IFN-beta 1b were found to be significantly higher than that of other groups. These results support that the ability of inducing anti-inflammatory cytokine IL-10 plays a role in the clinical advantage of IFN-beta 1b in MS treatment.

Two-day induction with thymoglobulin in kidney transplantation: risks and benefits.

INTRODUCTION: The aim of this study was to develop an induction protocol to reduce allograft rejection with fewer posttransplant infections and malignancies. METHODS: In this prospective randomized study, a T- and B-cell depletion protocol, consisting of IV thymoglobulin (ATG 5 mg/kg/d) plus methylprednisolone (500 mg/d) plus azathiopurine (2 mg/kg/d), was on days 0 and 1 after renal transplantation. CyA was introduced at day 3.39 among patients undergoing either primary living related (n = 16) or cadaveric (n = 23) transplants excluding recipients of full-HLA-matched sibling, or five- and six-HLA-matched cadaveric donor kidneys. The adequacy of immunosuppression was evaluated by flow cytometric analysis for total, CD3+ (T-cell), and CD19+ (B-cell) lymphocytes. RESULTS: The acute rejection rate was 6% and 37/39 patients are alive with functioning grafts at an average follow-up of 14.5 months. The overall patient and graft survival rate was 95%. Their mean creatinine value was 1.27 mg/dL. Six patients (16%) required hospitalization due to serious infections. The two deaths were attributed to septicemia and brain abcess caused by unusual agents, namely, Rhodococcus equi and Sporobolomyces. One patient presented with a cutaneous Kaposi sarcoma in the 11th month posttransplant. CONCLUSION: A Two-day induction protocol with thymoglobulin yields acceptable acute rejection rates among renal transplants. However, caution is necessary for adverse events, particularly atypical bacterial and fungal infections.

Exchange kidney transplantation: a good solution in living kidney transplantation.

OBJECTIVES: Kidney transplantation seems to be the best treatment modality for end-stage renal disease patients. But not every patient on the waiting list is able to find a kidney. To increase transplantations, centers have tried to find new options. MATERIAL AND METHODS: In the period of November 1994 through June 2004, among 265 renal transplantations, 182 (68.6%) were from living related donors, namely first- and second-degree relatives, spouses, or parents-in-law of the patients. Four patients, who did not have living related donors, had the opportunity of renal transplantation from living donors by exchanging their donors. RESULTS: All the kidneys functioned immediately. No complications and no acute rejection episodes were observed in the postoperative period up to 12 months. Serum creatinine levels were 1.9, 1.2, 1.6, and 2.4 mg/dL on postoperative day 7; 1.4, 1.0, 1.1, and 1.1 mg/dL at 1 month after transplantation; 1.5 and 1.2 mg/dL at month 6 after transplantation; 1.6 and 1.4 mg/dL at 1 year after transplantation. CONCLUSION: We believe that exchange kidney transplantations represent a good alternative for end-stage renal patients who do not have suitable close living related donors.

Uterine artery function in pregnant rats fed a diet supplemented with animal lard.

We hypothesised that maternal uterine artery vascular dysfunction could contribute to cardiovascular dysfunction in offspring of rats fed a diet rich in fat. Sprague-Dawley rats were fed for 10 days prior to pregnancy and throughout gestation either: (a) a control breeding diet, or (b) the same diet supplemented with 20 % w/w lard, vitamins, essential micronutrients and protein to control values. At 20 days gestation vascular function was assessed in uterine arteries and third-order mesenteric arteries. Vascular reactivity in response to application of potassium, noradrenaline, the thromboxane analogue U46619, acetylcholine and nitric oxide was assessed. Maternal plasma concentrations of factors likely to contribute to endothelial dysfunction were measured. Maximum acetylcholine-induced relaxation was impaired in the mesenteric arteries of the lard-fed dams (max % relaxation: lard-fed, 69.7 +/- 6.48; control, 85.37 +/- 2.69, P = 0.03). Uterine artery vascular function was similar in the two groups (max % acetylcholine-induced relaxation: lard-fed, 73.7 +/- 4.01; control, 77.5 +/- 4.72, P = 0.98). Concentrations of plasma lipids, 8-epi-PGF(2alpha) and leptin were normal, whereas insulin and corticosterone concentrations were raised in the lard-fed group (insulin (ng ml(-1)): lard-fed, 8.04 +/- 0.47; control, 1.35 +/- 0.37, P < 0.0001; corticosterone (ng ml(-1)): lard-fed, 1164.0 +/- 170.9; control, 541.9 +/- 96.3, P = 0.005). Fetal and placental weights were reduced in lard-fed dams (fetus (g): lard-fed, 4.27 +/- 0.38; control, 2.96 +/- 0.40, P = 0.025; placenta (g): lard-fed, 0.72 +/- 0.06; control, 0.57 +/- 0.04, P = 0.05). Cardiovascular dysfunction in offspring is not associated with reduced uterine artery endothelial function but is associated with activation of the hypothalamic-pituitary-adrenal axis, hyperinsulinaemia and fetoplacental growth retardation.

Protective effects of carnitine in an experimental ischemia-reperfusion injury.

BACKGROUND/AIMS: We aimed to determine the role of exogenous carnitine to prevent ischemia-reperfusion damage in liver tissue in experimental model. METHODS: Rats were divided into four groups as Sham (SG), 30% Hepatectomy (HG), ischemia-reperfusion +30% hepatectomy (IRHG) and ischemia-reperfusion+30% hepatectomy+carnitine (IRHCG). Serum AST, ALT and GGT levels have been determined in systemic blood samples (post-hepatic vena cava) and liver tissue and serum carnitine levels in blood samples from portal vein (pre-hepatic blood samples). RESULTS: Serum carnitine levels were significantly higher in IRHCG compared to SG (P < 0.01). Each of the serum AST, ALT and GGT levels were statistically higher in HG, IRHG and IRHCG than SG (P < 0.001). While these values in IRHG were also higher than those in HG (P < 0.001), in IRHCG enzyme levels were significantly lower than IRHG (P < 0.001). Liver tissue damage was less in IRHCG than IRHG statistically (P < 0.001). CONCLUSIONS: This animal model implies that exogenous carnitine supplementation may be helpful in preventing free oxygen radical damage and inflammatory reactions in liver tissue.

Intraleucocyte platelet-activating factor levels in desmopressin-treated patients with haemophilia A and von Willebrand disease.

Despite the intensive clinical use of 1-deamino-8-D-arginine vasopressin (desmopressin; DDAVP) for 20 years, its mechanism of action is still not completely explained. It has been proposed that DDAVP stimulates release of a 'second messenger' which in turn stimulates release of von Willebrand factor (vWF) from endothelial cells. Platelet-activating factor (PAF) and interleukin (IL)-6 were individually proposed to be mediators for haemostatic action. The aim of this study was to investigate cellular-based PAF levels in patients with haemophilia A (HA) and von Willebrand disease (vWD) before and after DDAVP treatment and also to look for any probable relationship between the haemostatic response of DDAVP and cellular PAF activities. In total, 20 patients (11 HA and nine vWD) were enrolled in the study. DDAVP was given subcutaneously as a single dose (0.3 microg kg(-1)). Ten patients responded to DDAVP and were defined as the 'able group' (four mild HA, six type 1 vWD). The remaining 10 patients did not respond to DDAVP and were defined as the 'unable group' (seven severe HA, three type 3 vWD). Released (extracellular) and intracellular (intraleucocyte) PAF levels under the stimulation of specific agents (A23187 and Zymosan) were measured by high-performance liquid chromatography and radioimmunoassay. Extracellular and intracellular PAF activities were not detected without stimulation in healthy children whereas significantly higher PAF levels were found in the patients (extracellular: 37.5 +/- 34.4 ng per 10(7) cells; intracellular: 24.8 +/- 23.5 ng per 10(7) cells; P=0.0001). Intracellular PAF levels obtained from in vitro unstimulated cells were significantly higher in DDAVP-responsive (able) patients in comparison to DDAVP-unresponsive (unable) patients (52.1 +/- 18.5 vs. 28.9 +/- 8.0 ng per 10(7)cells). After in vitro stimulation by A23187, intracellular PAF activities were significantly higher in patients than in controls (209.3 +/- 26.1 vs. 172 +/- 18.1 ng per 10(7) cells). Intracellular PAF levels obtained from in vitro stimulated cells by A23187 were also significantly higher in the 'able' patients in comparison to the 'unable' patients (277 +/- 43.5 vs. 225 +/- 30 ng per 10(7)cells). In conclusion, cellular PAF activities are significantly higher in patients with HA and vWD. We also suggest that PAF, especially intracellular PAF mediates intracellular signalling and may be one of the important mediators for the haemostatic response of DDAVP.

Levels of leukotriene B4 in gingival crevicular fluid and gingival tissue in specific periodontal diseases.

BACKGROUND: Leukotriene B4 (LTB4), a product of the lipoxygenase pathway of arachidonic acid metabolism, exhibits numerous activities that can account for most of the features of host responses seen in periodontal diseases. The aim of the present study was to examine the role of LTB4 in the pathogenesis of specific periodontal diseases. METHODS: LTB4 levels were investigated in gingival crevicular fluid (GCF) and gingival tissue (GT) samples of 10 patients with chronic periodontitis (CP), 12 patients with generalized aggressive periodontitis (GAgP), 6 patients with localized aggressive periodontitis (LAgP), 6 patients with gingivitis (G), and 6 periodontally healthy subjects (H). Periodontal status was evaluated by measuring probing depth, gingival index, papillary bleeding index, and plaque index. LTB4 was extracted from the samples by solid-phase method using C18 cartridge and was purified by high performance liquid chromatographic method and then analyzed by radioimmunoassay. RESULTS: All patient groups had significantly higher levels of GCF and GT LTB4 compared to the control group (P<0.005). The CP patients had the highest LTB4 levels compared to those in other patient groups (P<0.005). GAgP, LAgP, and G groups had similar amounts of GCF and GT LTB4 (P>0.005). When the data were expressed as concentration, the CP group was found to have higher concentration of LTB4, compared to that of control group (P<0.005). GAgP, LAgP, and G groups had similar LTB4 concentration compared to that of control group (P>0.005). No significant difference was found between GAgP, LAgP, and G groups (P>0.005). The CP group had higher LTB4 concentration compared to both GAgP and LAgP groups (P<0.005). Although the CP group had a higher GCF LTB4 concentration compared to G group, this difference did not reach significance (P>0.005). No significant correlation was found between GCF and GT LTB4 levels and clinical parameters. CONCLUSIONS: The results of the present study indicate that LTB4 is likely to be an important mediator in regulating inflammatory responses in the human periodontal tissues. This lipid mediator may play an important role in the pathophysiology of periodontal disease.

Is mannitol effective against platelet-activating factor (PAF)-induced liver damage in obstructive jaundice?

BACKGROUND/AIMS: Platelet-activating factor, is a unique phospholipid with a broad range of biological activities that may be relevant in the development of inflammatory reactions. Platelet-activating factor has been suspected to play an important role in liver pathophysiology. The cultured Kupffer and endothelial cells produce and release platelet-activating factor in order to facilitate communication between hepatic sinusoidal and parenchymal cells. In this study, in the experimental jaundice model, platelet-activating factor levels were measured in liver tissue and plasma and the possible effects of mannitol on this mediator were assessed. METHODOLOGY: The experimental model consisted of 7 rats in the control group (CG), 7 rats in the sham operation group (ShG), and 7 rats in the obstructive jaundice group (JG) created by ligating the common bile duct. The last group was the mannitol-treated jaundiced group (MJG) and all animals in this group received 20% mannitol in doses of 2 mL/day, intraperitoneally, following common bile duct ligation. A week later all animals were sacrificed and plasma and liver tissue samples were collected. Platelet-activating factor levels were measured by radioimmunoassay technique. RESULTS: Liver tissue platelet activating factor levels (pg/mg tissue protein) were 72 +/- 18 in the CG, 183 +/- 51 in the JG, 84 +/- 17 in ShG, and 124 +/- 36 in MJG. Plasma levels were 460 +/- 13, 1600 +/- 40, 560 +/- 19, and 1200 +/- 23, respectively. In both sample types, MJG and JG values were significantly different from CG and ShG as well. MJG levels were also different from JG. CONCLUSIONS: These results showed that plasma and liver tissue platelet-activating factor levels are increased in experimental obstructive jaundice; and activation of this mediator contributes to the ongoing liver injury. Mannitol may improve or lessen this damage.

Aldehyde analysis by high performance liquid chromatography/tandem mass spectrometry.

This paper describes the development of a high performance liquid chromatography/tandem mass spectrometric (MS/MS) procedure for the specific qualitative and quantitative analysis of lipid aldehydes in biological matrices. A derivatisation method, which results in molecules that exhibit a common product ion on MS/MS, permits informative precursor ion scans, at high sensitivity. This has been applied to the examination of plasma in order to examine the production of aldehydes consequent on in vitro lipid oxidation. Quantitative analysis of target molecules using multiple reaction monitoring has been developed to permit quantitation in the same matrices.

Phenotypic and functional analysis of peripheral blood mononuclear cells in generalised aggressive and chronic periodontitis patients.

BACKGROUND: The aetiology and pathogenesis of periodontal disease may involve dysfunctions in the cellular immune responses. The aim of the present study was to study the phenotypic and functional activities of peripheral blood mononuclear cells (PBMC) from 16 patients with generalised aggressive periodontitis (G-AP), 13 patients with chronic periodontitis (CP) and 20 periodontally healthy subjects (H). METHODS: The relative counts of CD3+, CD4+, CD8+ T cells, T cells exhibiting HLA DR antigen and interleukin-2 receptor, CD19+ B cells and natural killer cells were determined by two colour flow cytometry using monoclonal antibodies. Blastogenic response of PBMC to mitogen phytohemagglutinin (PHA) was assessed after 96 h incubation by measuring 3(H)-thymidine incorporation and the results were expressed as net counts per minute. Spontaneous PBMC proliferation was also evaluated in unstimulated culture and the results were reported as mean counts per minute. Comparisons of G-AP, CP and H groups were performed using the Kruskal-Wallis and Bonferroni-corrected Mann-Whitney U test. RESULTS: No significant differences in the relative counts of PBMC subsets were found between G-AP, CP and H groups (P > 0.05) with the exception of lower relative amount of CD3+ T cells found in the CP group compared with healthy subjects (P = 0.0048). Blastogenic response to PHA was significantly suppressed in G-AP and CP groups relative to that of H group (P < 0.02). However, there was no significant difference in the blastogenic response to PHA between G-AP and CP groups (P > 0.05). Spontaneous proliferative response of G-AP and CP groups was also significantly lower compared to that of H group (P < 0.02). Similarly, no significant difference in spontaneous PBMC response was observed between G-AP and CP groups (P > 0.05). CONCLUSIONS: Although G-AP and CP patients have similar amount of immune cells compared to healthy subjects, reduced functional activities of these cells may suggest the existence of defective cellular immune mechanism for the susceptibility to periodontal disease. One has to keep in mind that periodontal diseases have a genetic basis and the present functional analysis might not be connected to the actual genetic predisposition to the disease.

Increase of the isoprostane 8-isoprostaglandin f2alpha in maternal and fetal blood of rats with streptozotocin-induced diabetes: evidence of lipid peroxidation.

OBJECTIVE: Pregnancy complicated by diabetes is associated with maternal complications and fetal abnormalities. Animal models of diabetes suggest that heightened free radical production may be implicated in the pathogenesis of this condition. The purpose of this investigation was to evaluate oxidative stress in plasma from diabetic rats and their fetuses through measurement of concentrations of 8-isoprostaglandin F(2alpha), a stable marker of lipid peroxidation. STUDY DESIGN: Diabetes was induced in virgin and pregnant rats with streptozotocin. Blood samples were collected after 20 days of diabetes. Adult and fetal plasma 8-isoprostaglandin F(2alpha) concentrations were determined by gas chromatography-mass spectroscopy. RESULTS: Significantly higher plasma 8-isoprostaglandin F(2alpha) concentrations were observed in the virgin rats with diabetes and in both the pregnant dams with diabetes and their fetuses when compared with their respective control groups without diabetes (P <.001). CONCLUSION: Oxidative stress was induced in both mother and fetus in rodent pregnancy complicated by diabetes. This finding may have implications for fetal dysmorphogenesis and in fetal programming for adulthood disease.

Raised saturated-fat intake worsens vascular function in virgin and pregnant offspring of streptozotocin-diabetic rats.

Adult offspring of severely diabetic pregnant rats are insulin resistant and display cardiovascular dysfunction. When pregnant they develop mild hyperglycaemia. Diets high in saturated fat have been implicated in the development of cardiovascular disease and vascular dysfunction. In the present study we have determined vascular function in small mesenteric arteries from offspring of normal (OC) and diabetic (OD) rats fed standard chow and offspring of diabetic rats fed a diet high in saturated fats (OD-HF) from weaning to adulthood, and throughout their subsequent pregnancies. OD rats displayed an increased sensitivity to noradrenaline (P < 0.05) and impaired sensitivity to the endothelium-dependent vasodilator, acetylcholine. The component of acetylcholine-induced relaxation attributable to endothelium-derived hyperpolarizing factor was reduced in OD-HF rats. Pregnant OD rats also demonstrated impaired maximum relaxation to acetylcholine (pregnant OD rats v. pregnant OC rats P < 0.05). In pregnant OD-HF rats noradrenaline sensitivity was enhanced and endothelium-dependent relaxation further reduced (pregnant OD-HF rats v. pregnant OC rats P < 0.001). The isoprostane, 8-epi-prostaglandin F2alpha, a marker of oxidative stress, was increased in pregnant OD rats (pregnant OD rats v. pregnant OC rats P

Levels of leukotriene B4 and platelet activating factor in gingival crevicular fluid in renal transplant patients receiving cyclosporine A.

BACKGROUND: Cyclosporine A (CsA) is a potent immunosuppressant effectively used to prevent organ transplant rejection and also to treat several systemic diseases. CsA-induced gingival overgrowth (CsA GO) is the most widely seen side effect of this drug; its pathogenesis is not completely understood. The aim of the present study was to identify the role of leukotriene B4 (LTB4) and platelet activating factor (PAF) in the pathogenesis of CsA GO. METHODS: LTB4 and PAF levels were detected in gingival crevicular fluid (GCF) samples from renal transplant patients receiving CsA therapy and exhibiting CsA GO, from patients with gingivitis and from periodontally healthy subjects. Plaque index, papilla bleeding index, and hyperplastic index were recorded at each study site. GCF samples and clinical data were obtained from: 2 sites exhibiting CsA GO (CsA GO+) and 2 sites not exhibiting CsA GO (CsA GO-) in each CsA-treated patient; 2 diseased sites in each patient with gingivitis; and 2 healthy sites in each subject with clinically healthy periodontium. LTB4 was extracted from the samples by solid-phase method using C18 cartridge and purified by high-performance liquid chromatographic (HPLC) method and analyzed by radioimmunoassay (RIA). PAF was extracted from GCF samples passing through amberlit resin columns, purified by HPLC, and analyzed by RIA. RESULTS: Total amounts of LTB4 and PAF in GCF were higher in CsA GO+ sites compared to the healthy sites from healthy controls. However, the amount of LTB4 and PAF elevation in CsA GO+ sites was not significantly higher than those in diseased sites. Clinical degrees of gingival inflammation were also similar between CsA GO+ and diseased sites. LTB4 and PAF total amounts in GCF were higher in CsA GO+ sites compared to CsA GO- sites in the same subjects, but this difference just failed to reach significance. Similar findings were obtained with concentration data. CONCLUSIONS: The results of this study indicate that CsA therapy does not have a significant effect on GCF LTB4 and PAF levels and that gingival inflammation seems to be the main reason for their elevation. In CsA-treated patients, alterations in LTB4 and PAF levels might play a role in CsA GO through some asyet unknown mechanism. To our knowledge, this is the first report describing the levels of lipid mediators in GCF of CsA-treated patients. We assume that further studies will contribute to the understanding of the pathogenesis of CsA-induced gingival overgrowth.

Platelet-activating factor concentrations in healthy and septic neonates.

Platelet-activating factor levels were measured in nine preterm infants with Klebsiella pneumonia septicaemia, eight healthy preterm infants of similar gestational age and ten healthy full-term infants of the same postnatal age at sampling. The platelet-activating factor levels of the healthy preterm and term groups did not differ significantly, but were elevated compared to the other two groups in the septic preterm infants (P < 0.01). Platelet-activating factor levels increase upon stimulation by Gram negative bacteraemia and are a important mediator of neonatal sepsis.

Cholesterol-independent endothelial dysfunction in virgin and pregnant rats fed a diet high in saturated fat.

1. Western diets high in saturated fat are associated with an increased incidence of cardiovascular diseases. In this study we have evaluated vascular endothelial function and oxidative stress in virgin rats fed a normal (VC) or high in saturated fat diet (VHF) (20 % lard and corn oil w/w) from weaning until adulthood, and throughout subsequent pregnancy (PC and PHF, respectively). 2. The saturated fat diet was associated with enhanced noradrenaline sensitivity in small mesenteric arteries from VHF rats (VHF vs. VC, P < 0.05) and blunted endothelium-dependent relaxation in VHF and PHF rats (VHF vs. VC, P < 0.001; PHF vs. PC, P < 0.05). Endothelial dysfunction was attributable to a reduced nitric oxide component of relaxation in VHF rats, and blunted prostacyclin and endothelium-derived hyperpolarizing factor components in PHF rats. 3. Other than plasma cholesterol, which was reduced in VHF and PHF rats, plasma lipids were normal. Fasting plasma insulin and glucose concentrations were raised in VHF rats (P < 0.05) and the plasma marker of oxidative stress, 8-iso PGF2alpha, was increased in PHF animals (P < 0.01). 4. These findings suggest that endothelial dysfunction induced by a saturated fat diet is cholesterol independent and likely to be of different mechanistic origin in virgin and pregnant rats.

Increased gastric production of platelet-activating factor, leukotriene-B4, and tumor necrosis factor-alpha in children with Helicobacter pylori infection.

The concentrations of platelet-activating factor (PAF), leukotriene-B4 (LTB4), and tumour necrosis factor-alpha (TNF-alpha) in homogenate supernatants of gastric mucosal biopsy specimens and in gastric juice from Helicobacter pylori-positive (N = 21) and -negative children (N = 14) were investigated in order to determine whether these lipid mediators and the cytokine are involved in the inflammatory reaction of H. pylori-associated gastritis. PAF and LTB4 concentrations were measured after high-performance liquid chromatography (HPLC) purification by specific radioimmunoassay, and TNF-alpha concentrations were determined by using an enzyme-linked immunosorbent assay. The concentrations of PAF, LTB4, and TNF-alpha measured in gastric juice and biopsy homogenate supernatants of children with H. pylori-positive gastritis were found to be statistically elevated and in positive correlation with each other. This study suggested that increased local mucosal production of potent proinflammatory agents such as PAF, LTB4, and TNF-alpha may be implicated in the pathogenesis of H. pylori-associated gastritis in childhood.

Is administration of n-3 fatty acids by mucosal enema protective against trinitrobenzene-induced colitis in rats?

We investigated the protective role of fish oil (FO-source of n-3 FA) enriched diet (in the first protocol) in 20 rats and FO administration intrarectally (in the second protocol) in 40 rats with trinitrobenzene (TNB) colitis. All colonic specimens were pathologically evaluated, myeloperoxidase enzyme activities were measured, leukotriene B4 (LTB4) and LTC4 levels were determined by radioimmunoassay. In the first protocol 10 rats (group A1) were fed with 8% sunflower and cotton oil enriched diet and (group A2) with 8% FO enriched diet for 6 weeks. At the end of this period, TNB (30 mg in 0.25 ml of 30% ethanol) were intrarectally administered. After 2 weeks, rats were sacrificed. MPO activities (2.47 versus 30.17), LTB4 (34.5 versus 903.3) and LTC4 (77.7 versus 456.0) levels were significantly reduced in group A2 compared with group A1 (P<0.005). There was also a significant difference in pathologic scores (1.55 versus 2.12, P<0.002) between two groups. In the first part of the second protocol, 20 male rats were randomized into two equal groups (B1 and B2) and TNB colitis was induced. After 1 day, 1 ml of saline (group B1) or n-3 FA enemas (group B2) were administered every day for 2 weeks. At the end of this period, rats were sacrificed and evaluated as done for previous groups. Although there was no significant difference between the two groups in comparison with MPO enzyme activities and pathologic scores, the LTB4 (130.1 versus 971.0) and LTC4 (126.0 versus 532.0) levels of FO group were significantly reduced (P<0.005). In the second part of the second protocol, 20 male rats were randomized into two groups. One millilitre of saline (group B3) or FO enemas (group B4) were administered to rats every day for 3 days. At the fourth day, TNB-colitis was induced and after 24 h rats were sacrificed. We could not find any significant difference in MPO activities, pathologic scores, LTB4 and LTC4 levels between groups B3 and B4. In conclusion, FO enriched diet decreased both pathologic damage and tissue LT levels. The second protocol of our study revealed that the long-term FO enemas decreased the LTB4 and LTC4 levels; however, did not have any beneficial effect on the tissue lesions. Short periods of FO enemas did not have a protective role in the occurrence of experimental colitis. The present study showed that FO enemas significantly decreased LT levels. The protective effect of FO (oral and enema) in TNB colitis may open a new insight into the treatment of inflammatory bowel disease.

Increased gastric juice leukotriene B4, C4 and E4 concentrations in children with Helicobacter pylori colonization.

During recent years, the role of inflammatory lipid mediators in the pathophysiology of Helicobacter pylori (H. pylori) infections has been investigated in several studies. The concentrations of leukotrienes (LTs) in gastric juice from H. pylori positive (n = 13) and negative (n = 18) children with recurrent abdominal pain were studies in order to determine whether these lipid inflammatory mediators are involved in local and systemic biological actions. Gastric juice samples and biopsy specimens of mucosa were obtained endoscopically from 31 patients with recurrent abdominal pain for assessment of LTs and histopathological examination. In this study, all children with recurrent abdominal pain were investigated by rapid urease test and histological assessment for H. pylori colonization. Leukotriene levels were measured by high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) in gastric juice samples. Gastric juice LTB4, LTC4, and LT4 levels were significantly higher in patients with H. pylori colonization than in children without H. pylori colonization. These results indicate that increased gastric content of proinflammatory mediators (LTB4, LTC4, and LT4) may be related to the pathogenesis of H. pylori-associated gastritis.