Butanol-extracted lipoteichoic acid induces in vivo leukocyte adhesion

Lipoteichoic acid (LTA), an immunostimulatory component of the cell walls of Gram positive bacteria, has pro-inflammatory effects in vitro and in vivo. However, one in vivo study concluded that LTA had no noticeable effects on leukocyte recruitment. In this study we investigated the effects of highly purified LTA, prepared by butanol extraction (Bu-LTA) at room temperature, on in vivo leukocyte adhesion. Using intravital microscopy we measured adhesion of leukocytes in mesenteric post-capillary venules of rats and mice. Topical superfusion of Bu-LTA (1 μg/ml) in rats significantly (p < 0.05) increased adhesion within 30 min. By contrast, hot phenol-extracted LTA did not increase adhesion. Alkaline hydrolysis of Bu-LTA removed alanine residues and prevented adhesion. Also, pre-administration of anti-rat β₂-integrin antibody abolished Bu-LTA-induced adhesion. Finally, intraperitoneal injection of Bu-LTA (100 μg/ml) into mice also significantly (p < 0.01) increased leukocyte adhesion measured at 60 min. In conclusion, Bu-LTA with intact alanine residues promotes β₂-integrin-dependent leukocyte adhesion in vivo.     

Peptide mimotopes of complex carbohydrates in Salmonella enterica serovar typhi which react with both carbohydrate-specific monoclonal antibody and polyclonal sera from typhoid patients

Polyclonal sera from typhoid patients and a monoclonal antibody, mAb ATVi, which recognizes the capsular polysaccharide Vi antigen (ViCPS), were used to select for peptides that mimic the ViCPS by using a phage-displayed random 12-mer peptide library. Two major common mimotopes selected from the library carried the amino acid sequences TSHHDSHGLHRV and ENHSPVNIAHKL. Enzyme-linked immunosorbent assays (ELISAs) showed that these peptides carry mimotopes to ViCPS. Phage clones that contained the 12-mer peptides were also tested against pooled/individual typhoid patients' sera and found to have 3 to 5 times higher binding compared to normal sera. By using Phage-ELISA assays, the derived synthetic peptides, TSHHDSHGLHRV and ENHSPVNIAHKL, were tested against a monoclonal antibody mAb ATVi and over 2-fold difference in binding was found between these peptides and a control unrelated peptide, CTLTTKLYC. Inhibition of the mAb's binding to ViCPS indicated that the synthetic peptides successfully competed with the capsular polysaccharide for antibody binding.     

Effect of bilirubin on the fungicidal capacity of human neutrophils

The effect of unconjugated bilirubin on the fungicidal capacity of human neutrophils was examined. Per cent Torulopsis glabrata killed in the presence of 1 x 10(-5) and 5 x 10(-5) M bilirubin was 72.7 +/- 2.5 and 40.9 +/- 7.2 respectively, compared to 81.4 +/- 4.1 in control (p less than 0.01 and p less than 0.001), respectively). This effect was not due to a concomitant inhibition of phagocytosis. The results suggest that jaundiced neonates may be more susceptible to fungal infections.     

Prevalence of intestinal parasite among patients attending two hospitals in French Guiana: A 6-year retrospective study

IntroductionIntestinal parasitic diseases are a global health problem. Due to its equatorial climate, vast territory with isolated areas and the precariousness of its population, intestinal parasitosis is considered to be a major issue in French Guiana but only few data are available and these mainly focus on specific population. We aimed at determining the parasitic index and at describing the characteristics of these infections in order to develop preventive strategies.Material and methodsWe retrospectively analysed all the parasitological samples recorded in the register of the two main laboratories of French Guiana between 2011 and 2016. The parasitic index was the percentage of parasitised patients in comparison with the total number of subjects studied. A patient who underwent several positive parasitological examinations was considered only once in the analysis at the time of the first sampling.ResultsA total of 15,220 parasitological samples of 9,555 patients were analysed and 2,916 were positive in 1,521 patients. The average infestation rate and parasitic index were 19.2% and 16.0%, respectively. The parasitic index remained stable between 2011 (18.2%) and 2016 (18.3%). The patients were mainly men (66.4%), with a median age of 33.0 years (26.3% of patients were under 18 years of age) and lived mainly in the Central Agglomeration (48.2%) and in West Guiana (37.4%). Hookworms were the most common parasite (25.2%) followed by Entamoeba coli (13.3%), Strongyloides stercoralis (10.9%) and Giardia intestinalis (10.8%). Among the infected patients, 31.0% presented mixed infections and 67.5% of them had at least one pathogenic parasite. The patients aged from 0 to 18 years presented significantly more polyparasitism (30.9%) than monoparasitism (24.3%, p<0.001). Ancylostoma sp and Strongyloides stercoralis were mainly diagnosed during the rainy season (59.5% and 64.7% respectively), in men (78.6% and 81.1% respectively) and in patients aged from 18 to 65 years (86.6% and 76.6% respectively) whereas, Giardia intestinalis infected mostly children under 5 years (59.5%) of age.ConclusionAlthough it may not be representative of the entire Guyanese population, the parasitic index remained high and stable from 2011 and 2016 and it justifies the need for an active prevention program as it was already done in the other French overseas departments such as Martinique and Guadeloupe.     

Flexible programs for the prediction of average amphipathicity of multiply aligned homologous proteins: application to integral membrane transport proteins

Simple flexible programs (TREEMOMENT and PILEUPMOMENT) are described for depicting the average amphipathicity (hydrophobic moment) along multiply aligned sequences of a family of evolutionarily related proteins. The programs are applicable to any number of aligned sequences and can be set for any desired angle corresponding to a residue repeat unit in a protein secondary structural element such as 100 per residue for an alpha- helix or 180 per residue for a beta-strand. These programs can be used to identify amphipathic regions common to the members of a protein family. The use of these programs is exemplified by showing that some families of integral membrane transport proteins (i.e. permeases of the bacterial phosphotransferase system (PTS) and the anion exchangers of animals) exhibit strikingly amphipathic alpha-helical structures immediately preceding the first hydrophobic transmembrane segment of their membrane-embedded domain(s). Other families, such as the major facilitator superfamily of uniporters, symporters and antiporters, do not exhibit this structural feature. The amphipathic structures in PTS permeases have been implicated in membrane insertion during biogenesis.     

Caffeine-induced impairment of glucose tolerance is abolished by beta-adrenergic receptor blockade in humans.

The caffeine-induced impairment of insulin action is commonly attributed to adenosine receptor (AR) antagonism in skeletal muscle. However, epinephrine, a potent inhibitor of insulin actions, is increased after caffeine ingestion. We tested the hypothesis that the insulin antagonistic effects of caffeine are mediated by epinephrine, and not by AR antagonism, in seven healthy men. On four separate occasions, they received 1) dextrose (placebo, PL), 2) 5 mg/kg caffeine (CAF), 3) 80 mg of propranolol (PR), and 4) 5 mg/kg caffeine + 80 mg of propranolol (CAF + PR) before an oral glucose tolerance test (OGTT). Blood glucose was similar among trials before and during the OGTT. Plasma epinephrine was elevated (P < 0.05) in CAF and CAF + PR. Areas under the insulin and C-peptide curves were 42 and 39% greater (P < 0.05), respectively, in CAF than in PL, PR, and CAF + PR. In the presence of propranolol (CAF + PR), these responses were similar to PL and PR. These data suggest that the insulin antagonistic effects of caffeine in vivo are mediated by elevated epinephrine rather than by peripheral AR antagonism.     

Biohydrogen production from wheat straw hydrolysate by dark fermentation using extreme thermophilic mixed culture

Hydrolysate was tested as substrate for hydrogen production by extreme thermophilic mixed culture (70°C) in both batch and continuously fed reactors. Hydrogen was produced at hydrolysate concentrations up to 25% (v/v), while no hydrogen was produced at hydrolysate concentration of 30% (v/v), indicating that hydrolysate at high concentrations was inhibiting the hydrogen fermentation process. In addition, the lag phase for hydrogen production was strongly influenced by the hydrolysate concentration, and was prolonged from approximately 11 h at the hydrolysate concentrations below 20% (v/v) to 38 h at the hydrolysate concentration of 25% (v/v). The maximum hydrogen yield as determined in batch assays was 318.4 ± 5.2 mL‐H₂/g‐sugars (14.2 ± 0.2 mmol‐H₂/g‐sugars) at the hydrolysate concentration of 5% (v/v). Continuously fed, and the continuously stirred tank reactor (CSTR), operating at 3 day hydraulic retention time (HRT) and fed with 20% (v/v) hydrolysate could successfully produce hydrogen. The hydrogen yield and production rate were 178.0 ± 10.1 mL‐H₂/g‐sugars (7.9 ± 0.4 mmol H₂/g‐sugars) and 184.0 ± 10.7 mL‐H₂/day L_reactor (8.2 ± 0.5 mmol‐H₂/day L_reactor), respectively, corresponding to 12% of the chemical oxygen demand (COD) from sugars. Additionally, it was found that toxic compounds, furfural and hydroxymethylfurfural (HMF), contained in the hydrolysate were effectively degraded in the CSTR, and their concentrations were reduced from 50 and 28 mg/L, respectively, to undetectable concentrations in the effluent. Phylogenetic analysis of the mixed culture revealed that members involved hydrogen producers in both batch and CSTR reactors were phylogenetically related to the Caldanaerobacter subteraneus, Thermoanaerobacter subteraneus, and Thermoanaerobacterium thermosaccharolyticum. Biotechnol. Bioeng. 2010;105: 899–908. © 2009 Wiley Periodicals, Inc.     

Effect of quinidine on mitogen-induced human lymphocyte transformation

The effect of quinidine on mitogen-induced lymphocyte transformation was examined using a microculture technique. The results showed that, at concentrations attainable during therapy, this drug exerted an inhibitory effect on lymphocyte proliferation. This inhibitory effect could be partially reversed by washing. The inhibitory effect was quite substantial when the drug was added 48 hours after the initiation of cultures. The clinical implications of this immunosuppressive property of quinidine are discussed.