Cytokines and sudden infant death.

BACKGROUND: It has been hypothesised that inflammatory reactions could play an important role in the pathway(s) leading to sudden and unexpected death in infancy. On a molecular level, these reactions are regulated by various cytokines. METHODS: To characterise the role of IL-1ß, IL-6 and TNFα more precisely, the concentrations of these cytokines were determined quantitatively using specific ELISA techniques in serum and cerebrospinal fluid (CSF) in 119 cases of sudden infant death. The infants were grouped into four categories (SIDS, SIDS with infection, natural death due to infection and unnatural death). RESULTS: A good correlation was found between CSF and serum for IL-6 (Spearman correlation coefficients (SCC), 0.73) and also for TNFα (SCC, 0.57), although the CSF concentrations were lower than that from the serum. There were no significant differences between the categories of death for any of the serum or CSF cytokines. Compared with normal values, increased serum concentrations of IL-1ß, IL-6 and TNFα were found in 70%, 69% and 38% of the cases respectively, indicating possible agonal or post-mortem changes of cytokine concentrations. In three cases very high cytokine concentrations were found (mainly for IL-6). This may have contributed to the mechanism of death (cytokine storm) in two of the cases. CONCLUSIONS: In a small group of patients, very high cytokine concentrations are a possible explanation for the cause of death ("cytokine storm").

Real-time quantitative PCR assay for the detection of Helicobacter pylori: no association with sudden infant death syndrome.

Helicobacter pylori infection is known to be one of the most common chronic infectious diseases in humans. Recently, a hypothesis was proposed that H. pylori infection could be a frequent cause for sudden infant death syndrome (SIDS). We have investigated this postulated association by examining formalin-fixed paraffin-embedded gastric tissues of a retrospective cohort of 94 SIDS cases: The presence of H. pylori was inferred from a newly developed real-time quantitative PCR assay with SYBR Green I detection. This assay is based on the amplification of the single-copy H. pylori-specific glmM gene. Accuracy and precision were verified using a plasmid containing a 977-bp fragment of this glmM gene. The assay was very sensitive, and as few as 30 template copies per PCR reaction could be detected even in the presence of excess human DNA. The assay was validated on mucosal biopsy samples of patients with known H. pylori infections. Interfering effects due to SIDS gastric tissue were excluded. Only two (2.1%) of the SIDS samples yielded H. pylori DNA copy numbers and only beyond the lowest standard concentration. These results could be confirmed independently by immunohistochemistry using an H. pylori-specific antibody. Thus, an infection by H. pylori is very rare in cases of SIDS, and thus the postulated association of H. pylori infection and SIDS cannot be confirmed.

Beta-ketoacyl-acyl carrier protein synthase IV: a key enzyme for regulation of medium-chain fatty acid synthesis in Cuphea lanceolata seeds.

With the aim of elucidating the mechanisms involved in the biosynthesis of medium-chain fatty acids in Cuphea lanceolata Ait., a crop accumulating up to 90% decanoic acid in seed triacylglycerols, cDNA clones of a beta-ketoacyl-acyl carrier protein (ACP) synthase IV (clKAS IV, EC 2.3.1.41) were isolated from C. lanceolata seed embryos. The amino acid sequence deduced from clKAS IV cDNA showed 80% identity to other plant KAS II-type enzymes, 55% identity towards plant KAS I and over 90% towards other Cuphea KAS IV-type sequences. Recombinant clKAS IV was functionally overexpressed in Escherichia coli, and substrate specificity of purified enzyme showed strong preference for elongation of short-chain and medium-chain acyl-ACPs (C4- to C10-ACP) with nearly equal activity. Further elongation steps were catalysed with distinctly less activity. Moreover, short- and medium-chain acyl-ACPs exerted a chain-length-specific and concentration-dependent substrate inhibition of clKAS IV. Based on these findings a regulatory mechanism for medium-chain fatty acid synthesis in C. lanceolata is presented.