Immediate diagnosis of ventriculits: evaluation of parameters independent of microbiological culture.

BACKGROUND: Generally accepted reference values in CSF diagnostics are not valid in cerebrospinal fluid (CSF) containing large amounts of blood. Residual blood may obscure ventriculitis as diagnostics largely depend on parameters such as cell count, lactic acid and total protein measurement. We sought to improve the diagnostics by evaluating a cytokine panel and soluble CD62L as markers of ventriculitis. In addition, we tested an algorithm of established parameters to predict ventriculitis in a specific patient collective. METHODS: Analysis was performed on ventricular CSF samples from 50 consecutive patients. Gram staining, microbiological culture, total cell count, total protein and CD62L expression on neutrophil granulocytes were analysed immediately. Cytokines and soluble CD62L were measured by flow cytometry. FINDINGS: Positive culture was detected in ten patients. Of all parameters tested only IL1-beta, IL8 and CD62L on neutrophils were significantly different between culture-positive and -negative patients. The highest predictive value was obtained when analysing IL1-beta. The predictive value of a parameter combination (IL6 in CSF, C-reactive protein and leukocytes in periphereal blood) was comparable to IL1-beta. Half of the patients in this analysis were identified as culture positive because of the lack of inflammatory response. CONCLUSIONS: IL1-beta and perhaps also IL8 provide very good analytical performance when looking for ventriculitis in patients with residual blood in CSF. Turn-around time is short, and results could be reported within 1 h for 24 h a day. In some patients application of glucocorticoids may result in restricted inflammatory response. Even in these patients IL1-beta provides a reliable parameter for the immediate diagnosis of ventriculitis.

CD62L on neutrophil granulocytes, a useful, complementary marker for the prediction of ventriculitis in blood-containing CSF.

OBJECTIVES: Presence of residual blood is a common problem in cerebrospinal fluid (CSF) diagnostics of ventriculitis. We hypothesised that neutrophil granulocytes in infected, blood-containing CSF lose CD62L expression. Therefore CD62L expression on neutrophils may present a complementary marker to distinguish between patients with residual blood and infection. DESIGNS AND METHODS: Evaluation was performed in 64 ventricular CSF samples sent to the laboratory for diagnostic investigation. Cell count, microbiological culture, total protein and flow cytometric analysis of CSF were performed. RESULTS: Cell counts and CD62L expression were significantly different between the culture positive and negative group. ROC-analysis revealed a significant predictive value for cell count and CD62L expression. Optimal cut-offs were calculated and a decision tree was established to predict a positive culture. CONCLUSIONS: Cell count and CD62L expression were predictive for a positive culture and the combination helped to increase specificity and sensitivity for the detection of ventriculitis in blood-containing CSF.

Dietetic effects of oral intervention with mare's milk on the Severity Scoring of Atopic Dermatitis, on faecal microbiota and on immunological parameters in patients with atopic dermatitis.

In a double-blind, placebo-controlled crossover trial, 23 patients consumed 250 ml mare's milk or placebo for 16 weeks. The aim was to examine the effects of mare's milk on the characteristics of atopic dermatitis (AD), on faecal microbiota and on clinical and immunological parameters. The intensity of AD was examined using the Severity Scoring of Atopic Dermatitis (SCORAD) index. During the mare's milk period, the mean SCORAD value of patients (n=23; 17 females, 6 males) decreased from 30.1 to 25.3 after 12 weeks (P<0.05) and to 26.7 after 16 weeks (P<0.1). In a subgroup (n=7) the SCORAD index and especially the pruritus decreased by 30% through the mare's milk period (P<0.01). In this subgroup, the faecal bifidobacteria increased during the mare's milk period from 4.6% to 11.9% of eubacteria (P<0.05). The immunological parameters, except C-reactive protein, were unchanged.

Long-term moderate intervention with n-3 long-chain PUFA-supplemented dairy products: effects on pathophysiological biomarkers in patients with rheumatoid arthritis.

n-3 long-chain PUFA (n-3 LC-PUFA) may improve cardiovascular and inflammatory diseases. The effects of n-3 LC-PUFA-supplemented dairy products on inflammation and immunological parameters, biomarkers of oxidative stress, serum lipids, and on disease activity were determined in patients with rheumatoid arthritis (RA). Forty-five subjects (forty-three females and two males) were randomly divided into two groups in a double-blind, placebo-controlled cross-over study. Both groups received placebo or verum products consecutively for 3 months with a 2-month washout phase between the two periods. Blood samples were taken at the beginning and at the end of each period. The dairy products generally improved serum lipids by increasing HDL and lowering lipoprotein a. The n-3 LC-PUFA supplements act to lower TAG. Additionally, a decreased lipopolysaccharide-stimulated cylo-oxygenase-2 expression was found in patients who had consumed the enriched dairy products. The majority of the CD analysed were not influenced, although n-3 LC-PUFA did suppress the immune response as lymphocytes and monocytes were found to be significantly decreased. The n-3 LC-PUFA did not increase the biomarkers of oxidative stress such as 8-iso-PGF(2alpha) and 15-keto-dihydro PGF(2alpha), and DNA damage like 7,8-dihydro-8-oxo-2'-deoxyguanosine. The long-term consumption of dairy products (2 x 12 weeks) diminished the excretion of hydroxypyridinium crosslinks, and favoured the diastolic blood pressure. The consumption of moderate doses of n-3 LC-PUFA in combination with dairy products did not improve the disease activity. However, there is evidence of cardioprotective effects. Furthermore, the long-term consumption of dairy products acts against the cartilage and bone destruction in RA.

Chlamydia pneumoniae infection of aortic smooth muscle cells reduces platelet-derived growth factor receptor-beta expression.

Chlamydia pneumoniae infection may play a role in the pathogenesis of atherosclerosis. In this study, an oligonucleotide microarray was utilized to examine the transcriptional response of human aortic smooth muscle cells (AoSMC) to C. pneumoniae infection. Alteration of mRNA expression in 71 out of 780 genes was detected at 24 h after infection. Among the down-regulated genes, platelet-derived growth factor receptor-beta (PDGFR-beta) was identified as a target for further analysis because the PDGF system is involved in the fibroproliferative response of SMC in atherogenesis. Reverse transcriptase PCR analysis demonstrated that C. pneumoniae inhibits the up-regulation of PDGFR-beta mRNA occurring in AoSMC after mock infection. PDGFR-beta protein synthesis was examined by immunoblotting and fluorescence-activated cell sorting. Compared with mock-infected cells, the amount of receptor protein was reduced at 24, 48, and 72 h after infection. Diminished PDGFR-beta synthesis in infected cultures was accompanied by the suppression of AoSMC growth following PDGF-BB stimulation. The interference of C. pneumoniae with PDGFR-beta expression may result in decreased SMC proliferation in atherosclerotic plaques, thereby affecting the development and stability of advanced lesions.

Dietary supplementation with trans-11- and trans-12-18 : 1 increases cis-9, trans-11-conjugated linoleic acid in human immune cells, but without effects on biomarkers of immune function and inflammation.

Trans-fatty acid intake is associated with an increased risk of CHD and diabetes. The effects of single trans-fatty acid isomers are largely unexplored. The present study examined the effects of a 6-week supplementation with two trans-18 : 1 isomers (trans-11 and trans-12) in human subjects on immune cells, several inflammatory and immunological biomarkers (for example, IL, TNFalpha, C-reactive protein, adiponectin, intercellular adhesion molecule-1, prostacyclin, phagocytic process). Following a 2-week adaptation period without supplements, the test group (n 12) received vaccenic acid (trans-11-18:1) and trans-12-18 : 1 in equal amounts (6.0 g/d) for 6 weeks. The control group (n 12) consumed an oil without trans-fatty acids and conjugated linoleic acids (CLA). Samples were collected at the end of both periods. Trans-11- and trans-12-18 : 1 were significantly increased in cellular lipids. The endogenous synthesis of cis-9, trans-11-CLA from trans-11-18 : 1 was demonstrated via increased CLA in cellular lipids of the test group. Generally, trans-isomer supplementation did not affect either inflammatory biomarkers (for example, IL-6, IL-8, TNFalpha) or immune function (for example, phagocytosis) during the present study. The dietary supplementation of trans-11- and trans-12-18 : 1 (6 g/d) and their accumulation in leucocytes had no effects on biomarkers of inflammation and immune function. However, because of the limited data on the safety of trans-fatty acid intake and effects of individual trans isomers on human health (for example, trans-9-18 : 1, trans-10-18 : 1) at present, it is prudent to reduce trans-fat intake in general.

Influence of prebiotics and antioxidants in bread on the immune system, antioxidative status and antioxidative capacity in male smokers and non-smokers.

Interest in functional foods is increasing. The aim of the present study was to investigate breads supplemented with functional components. One was bread supplemented with inulin, linseed and soya fibre (prebiotic bread). The other was a prebiotic antioxidant bread (pre-aox-bread), which additionally contained green tea powder, herbs and tomato paste. The effects of these two breads on immunological and antioxidative parameters were compared with control bread (placebo). Twenty smokers and eighteen non-smokers were enrolled in the randomised parallel study, which consisted of a control period and an intervention period, each lasting for 5 weeks. Daily intake of bread and nutrients did not differ between the intervention and the control period. Most of the twenty-three investigated immunological parameters measured in peripheral blood were unaffected. However, the percentage of CD19 increased after intervention with prebiotic bread, whereas intercellular adhesion molecule-1 (ICAM-1) and CD3+NK+ (P < 0.05) decreased in both intervention arms. The ferric reducing ability of plasma (FRAP) was increased after consumption of the pre-aox-bread for non-smokers (1256 v. 1147 micromol/l; P = 0.019) and remained unchanged for smokers consuming the pre-aox-bread. All analysed carotenoids (P

Flow cytometry as a diagnostic tool for hereditary spherocytosis.

Flow cytometric analysis of eosin-5'-maleimide-labeled red blood cells has been proposed as a new method of identifying hereditary spherocytosis (HS). The aim of the present study was to analyze sensitivity and specificity of this method. Red blood cells from patients with HS (n = 58) revealed significantly lower mean channel fluorescence values than red blood cells from normal subjects (n = 110), unaffected HS family members (n = 8), and patients with other anemias (n = 44). Taking a mean channel fluorescence of 400.0 units as the threshold value identified by logistic regression, sensitivity and specificity of the test for HS were 96.6 and 99.1%, respectively. Flow cytometric analysis is a valuable screening test for the diagnosis of HS.

Effects of propofol vs methohexital on neutrophil function and immune status in critically ill patients.

PURPOSE: Patients with severe brain injury often require long-term sedation and have a high incidence of nosocomial infections, causing an increased mortality rate. However, whether anesthetic drugs might contribute to immunosuppressive effects remains unclear. METHODS: In this prospective study, we investigated the effects of propofol (4-6 mg x kg(-1) x h(-1)) and methohexital (1-3 mg x kg(-1) x h(-1)) on neutrophil leukocyte function and immune status in 21 patients with brain injury who either received propofol (n = 12; 9 male, 3 female; mean age, 51 +/- 15 years) or methohexital (n = 9; 8 male, 1 female; mean age, 48 +/- 17 years) after admission to the intensive care unit (ICU). Both sedatives were administered over 7 days and individual dosage was adapted according to clinical requirements. Neutrophil leukocyte function was assessed as phagocytosis and respiratory oxidative burst activity. Furthermore, leukocyte subpopulations, and surface markers of lymphocytes and monocytes (CD3; CD4; CD45RO; CD4/CD45RO; CD25; CD4 and CD25; CD54; CD69; CD14/HLA-DR; CD8; CD3/HLA-DR; CD4 : CD8 ratio) were assessed. Blood samples were drawn on ICU admission, and on days 3, 7, and 14. Patients' demographics were compared by Wilcoxon test and laboratory results were compared by analysis of variance (ANOVA) for repeated measurements, with an all pairwise multiple comparison procedure. RESULTS: There were no significant differences in neutrophil oxidative burst and phagocytosis within or between the two groups at the different time points. With respect to cellular markers of lymphocytes and monocytes, all values throughout remained in the normal range. CONCLUSION: Methohexital and propofol exhibited no significant effects on neutrophil function and immune status in patients with severe brain injury requiring long-term sedation.

Role of interferon-stimulated gene factor 3gamma and beta interferon in HLA class I enhancement in synovial fibroblasts upon infection with Chlamydia trachomatis.

Chlamydia trachomatis infection can cause reactive arthritis that is associated with the persistence of chlamydial organisms in the joint. Fibroblasts of the synovial membrane represent host cells for Chlamydia during articular infection. In this study we investigated the expression of HLA class I molecules in synovial fibroblasts following infection with C. trachomatis D. The expression of HLA class I heavy chain (HLA-I) was up-regulated in infected cultures as shown by reverse transcription-PCR and immunoblotting. The increase in cell surface expression of HLA-I and beta(2) microglobulin on infected fibroblasts was demonstrated by flow cytometric analysis. Suppression of enhanced production of interferon-stimulated gene factor 3gamma (ISGF3gamma) in infected cell cultures by antisense oligonucleotide treatment reduced the level of HLA-I. Blocking antibodies to beta interferon (IFN-beta) inhibited the Chlamydia-induced enhancement of both ISGF3gamma and HLA-I. These findings show that the up-regulation of HLA-I in synovial fibroblasts infected with C. trachomatis is caused by the induction of IFN-beta, which in turn stimulates the synthesis of ISGF3gamma, a transcription factor participating in the regulation of the HLA-I gene. The IFN-beta-mediated expression of HLA-I on Chlamydia-infected cells may be a regulatory factor in the immune response in chlamydial infections.