The synergistic effect of FC gamma receptor IIa and interleukin-10 genes on the risk to develop systemic lupus erythematosus in Thai population

Several linkage analyses have consistently shown that systemic lupus erythematosus (SLE) susceptible genes are located on chromosome 1q21-44. In this study, two major candidate genes, interleukin-10 (IL-10) and Fc gamma receptor IIa (FcgammaRIIa), within these regions were investigated in Thai SLE patients. The genotyping of three single-nucleotide polymorphisms (promoter area: -1082, -819 and -592) within IL-10 gene and one polymorphism (change amino acid at position 131) within FcgammaRIIa gene was determined in 195 SLE patients and 159 ethnically matched controls. The RR/RH genotypes of FcgammaRIIa were found to be significantly increased in SLE patients compared with healthy controls [OR = 2.01, 95% confidence interval (CI) = 1.28-3.14, P= 0.001]. Interestingly, the synergistic effect between RR/RH genotypes of FcgammaRIIa and ACC/ACC haplotype of IL-10 in susceptibility to SLE was observed (OR = 7.84, 95% CI = 1.60-52.04, P= 0.002). In addition, the FcgammaRIIa, RR homozygotes was also strongly associated with anticardiolipin antibody production (OR = 6.09, 95% CI = 1.38-30.54, P= 0.006). The result demonstrated that ACC haplotype of IL-10 gene and FcgammaRIIa R131 polymorphism can be used as marker for genetic susceptibility and severity to SLE in Thai population, particularly individuals carrying both specific genotypes.     

Nasal absorption and local tissue reaction of insulin nanocomplexes of trimethyl chitosan derivatives in rats

Objectives The objective of this work was to explore the potential and safety of trimethyl chitosan (TMC) and PEGylated TMC for improved absorption of insulin after nasal administration. Methods The nasal absorption of insulin nanocomplexes of TMC or PEGylated TMC was evaluated in anaesthetized rats. Concomitantly, the histopathological effects of these nanocomplexes on rat nasal mucosa were studied using a perfusion fixation technique. Key findings All insulin nanocomplexes containing TMC or PEGylated TMC showed a 34–47% reduction in the blood glucose concentration, when the insulin absorption through the rat nasal mucosa was measured indirectly. In addition, the relative pharmacodynamic bioavailability (F_dyn) of the formulations was found to be dependent upon the charge ratio of insulin and polymer, regardless of polymer structure. The F_dyn apparently decreased with increasing charge ratio of insulin : polymer. Although acute alterations in nasal morphology by the formulations were affected by the charge ratio of insulin and polymer, the formulation of insulin/PEGylated TMC nanocomplexes was shown to be less toxic to the nasal epithelial membrane than insulin/TMC nanocomplexes. Conclusions PEGylated TMC nanocomplexes were a suitable absorption enhancer for nasal delivery of insulin.     

Comparison of two homogeneous HDL cholesterol methods in a large population study

OBJECTIVES: High-density lipoprotein cholesterol (HDL-C) is an independent risk factor for coronary heart disease. Data are lacking on the performance of homogeneous methods using a large number of samples. DESIGN AND METHODS: We compared the performance of two HDL-C direct assays, the Dimension RxL (the Dade method) and the COBAS INTEGRA (the Roche method), for population screening. Performance was assessed using 4214 sera obtained from the International Collaborative Study on Atherosclerosis and Stroke In Asia (InterASIA) participants. RESULTS: The method comparison results demonstrated that both methods were highly correlated (r = 0.972). Deming regression analysis showed a slope of 1.009 +/- 0.007, an intercept of 0.048 +/- 0.009 and a S(y/x) of 0.08. The means were 1.29 +/- 0.33 and 1.23 +/- 0.33 mmol/l for the Roche and Dade methods, respectively, and the range of observed values were 0.30-3.05 and 0.19-2.86 mmol/l, respectively. The 95% confidence interval for the mean of the method differences was -0.10 to 0.22 mmol/l. Percentages of low (> or = 1.55 mmol/l), normal (1.03-1.54 mmol/l), and high risk (< 1.03 mmol/l) results were 15.5, 55.4, 29.0 for the Dade and 19.3, 59.3, 21.4 for the Roche method. The percentage of concordantly classified subjects at each cut point was 77.1%, 84.4%, and 95.5%. The percentage of overall consistency subjects was 85.4%. Thirteen percent of subjects were discordantly classified into the higher-risk group while the 1.6% of subjects were discordantly classified into the lower-risk group. CONCLUSIONS: Both homogeneous HDL-C methods were correlated and agree well with one another. The percentage of concordantly classified subjects was high. Thus, either method is suitable for large population studies.     

A simple, rapid and sensitive detection of salmonella in food by polymerase chain reaction

A polymerase chain reaction (PCR) method was developed for detection of salmonella in food. A set of PCR primers was designed to amplify a 199 bp salmonella-specific DNA fragment derived from a repetitive DNA of Salmonella Weltevreden. The assay detected all 52 most prevalent serovars found in contaminated food in Thailand and no cross-reaction was observed with other non-salmonella organisms. The limit of detection was 1 fg of purified target DNA or five bacteria from pure culture. The detection of artificially contaminated food performed following a 6 h enrichment step was three bacteria per gram and the result was obtained within 4 h.     

Antilipopolysaccharide factor (ALF) of mud crab Scylla paramamosain: molecular cloning, genomic organization and the antimicrobial activity of its synthetic LPS binding domain

Antilipopolysaccharide factors (ALFs) are small basic proteins that can bind and neutralize lipopolysaccharide (LPS) and have broad spectrum antimicrobial activities. In this study, we describe the isolation of the full-length cDNA encoding for ALF peptide (ALFSp) of mud crab, Scylla paramamosain by sequencing a hemocyte cDNA library and using the rapid amplification cDNA end (RACE) method. A full-length ALFSp cDNA of 614 bp contains an open reading frame (ORF) of 372 bp, encoding 123 amino acid protein with 26 residues signal sequence. The calculated molecular mass of the mature protein is 11.18 kDa. The highly two conserve cysteine residues and putative LPS binding domain were observed in ALFSp peptide. Comparison of amino acid sequences revealed that ALFSp shared high identity with other known ALFs and had an overall similarity of 65, 64, 63, 61 and 59% to those of Fenneropenaeus chinensis, Litopenaeus vannamei, Marsupenaeus japonicus, Limulus polyphemus, and Tachypleus tridentatus, respectively. A neighbour-joining tree showed a clear differentiation of each species and also indicated that ALF from S. paramamosain, Carcinus maenas and Callinectes sapidus are closely related phylogenetically. The genomic DNA sequence of ALFSp gene consists of 1075 bp containing three exons and two introns. Tissue distribution analysis revealed that ALFSp was abundantly expressed in hemocytes, intestine, and muscle but not in eyestalk. The synthetic ALFSp peptide containing putative LPS binding domain revealed a strong antimicrobial activity against several bacteria especially on the growth of Gram-positive bacteria, Micrococcus luteus and Gram-negative bacteria, Vibrio harveyi suggested that ALFSp could play an essential role in defense mechanism in S. paramamosain.     

Natural history of plasma leakage in dengue hemorrhagic fever: a serial ultrasonographic study

BACKGROUND: Although plasma leakage is the major cause of mortality and morbidity in patients with dengue hemorrhagic fever (DHF), a detailed assessment of the natural course of this process is still lacking. We employed serial ultrasound examination to delineate the locations and the timing of plasma leakage and to evaluate the usefulness of ultrasound in detecting plasma leakage in DHF. METHOD: Daily ultrasound examinations of the abdomen and right thorax were performed in 158 suspected dengue cases to detect ascites, thickened gall bladder wall and pleural effusions. Cases were classified into dengue fever (DF), DHF or other febrile illness (OFI) based on serology and evidence of plasma leakage including hemoconcentration and pleural effusion detected by chest radiograph. RESULTS: Ultrasonographic evidence of plasma leakage was detected in DHF cases starting from 2 days before defervescence and was detected in some cases within 3 days after fever onset. Pleural effusion was the most common ultrasonographic sign of plasma leakage (62% of DHF cases one day after defervescence). Thickening of the gallbladder wall and ascites were detected less frequently (43% and 52% of DHF cases respectively) and resolved more rapidly than pleural effusions. The size of pleural effusions, ascites and gall bladder wall thickness in DHF grade I and II were smaller than those of grade III patients. Ultrasound detected plasma leakage in 12 of 17 DHF cases who did not meet the criteria for significant hemoconcentration. CONCLUSIONS: Ultrasound examinations detected plasma leakage in multiple body compartments around the time of defervescence. Ultrasonographic signs of plasma leakage were detectable before changes in hematocrits. Ultrasound is a useful tool for detecting plasma leakage in dengue infection.