Closing the 1,3-propanediol route enhances hydrogen production from glycerol by Halanaerobium saccharolyticum subsp. saccharolyticum

The effect of vitamin B₁₂ on hydrogen (H₂) and 1,3-propanediol (1,3-PD) production in glycerol fermentation of Halanaerobium saccharolyticum subsp. saccharolyticum was studied in batch cultivations. In addition, the effect of vitamin B₁₂ on glucose fermentation of H. saccharolyticum subsp. saccharolyticum and on glycerol fermentation of H. saccharolyticum subsp. senegalensis, that is a non-1,3-PD producing bacterium, and of Clostridium butyricum, that is known for vitamin B₁₂-independent 1,3-PD production, were studied for comparison. 1,3-PD production of H. saccharolyticum subsp. saccharolyticum was observed to be vitamin B₁₂-dependent and the H₂ production in glycerol fermentation was remarkably enhanced when 1,3-PD production was blocked by the lack of vitamin B₁₂ supplementation. The highest H₂ yield obtained by H. saccharolyticum subsp. saccharolyticum was 2.16 mol/mol glycerol. No significant effects of vitamin B₁₂ on metabolite production in glucose fermentation of H. saccharolyticum subsp. saccharolyticum or in glycerol fermentation of H. saccharolyticum subsp. senegalensis or C. butyricum were observed.     

Anaerobic on-site black water and kitchen waste treatment using UASB-septic tanks at low temperatures.

Anaerobic on-site treatment of black water (BW) and a mixture of black water and kitchen waste (BWKW) was studied in a two-phased upflow anaerobic sludge blanket septic tank (UASBst) at 10-20 degrees C. The processes were fed either continuously or discontinuously (twice per weekday). Moreover, BWKW was post-treated for nitrogen removal in an intermittently aerated moving bed biofilm reactor (MBBR) at 20 degrees C. Removal of total chemical oxygen demand (COD1) was efficient at minimum 90% with all three UASBst at all temperatures. Removal of dissolved COD (CODdis) was also high at approx. 70% with continuously fed BW and discontinuously fed BWKW, while with discontinuous BW feeding it was 20%. Temperature decrease had little effect on COD removals, though the need for phase 2 increased with decreasing temperature, especially with BWKW. Post-treatment of BWKW in MBBR resulted in approx. 50% nitrogen removal, but suffered from lack of carbon for denitrification. With carbon addition, removal of ca. 83% was achieved.     

A prospective, seroepidemiological study of the role of human papillomavirus in esophageal cancer in Norway

Infection with the human papillomavirus (HPV), notably HPV type 16, has been associated with esophageal cancer in seroepidemiological studies. To evaluate the consistency of the association, we performed a nested case-control study of HPV seropositivity and risk of esophageal cancer within a prospectively followed cohort of 300,000 Norwegian men and women who had donated blood samples to a serum bank. The data file of the serum bank was linked with the nationwide Cancer Registry of Norway to identify esophageal cancers diagnosed after donation of the serum sample. Fifty-seven cases and 171 matched controls were analyzed for antibodies to specific microorganisms, and odds ratios for developing esophageal cancer were calculated. There was an increased risk of developing esophageal cancer among HPV 16-seropositive subjects (odds ratio = 6.6; 95% confidence interval, 1.1-71) but not among Chlamydia trachomatis-seropositive subjects. Adjustment for the presence of serum cotinine, a marker of smoking habits, did not affect the estimates substantially. The seroepidemiological association between HPV 16 and esophageal cancer seems to be consistent in different countries.     

Whole blood production of thromboxane, prostacyclin and leukotriene B4 after dietary fish oil supplementation in man: effect of vitamin E

Antigenic peptides derived from several differentiation antigens of the melanocyte lineage were recently identified in human melanomas as targets for HLA-A2.1-restricted cytotoxic T lymphocytes (CTLs). To examine their potential role in tumour-directed immune responses in vivo, we determined CTL reactivity against seven antigenic peptides derived from the Melan A/MART-1, tyrosinase and gp100/Pmel17 antigens in the peripheral blood of 10 HLA-A2+ healthy controls and 26 HLA-A2+ melanoma patients. The influenza matrix peptide (GILGFVFTL) presented by HLA-A2.1 was used as a control peptide. CTL reactivity was assessed in a mixed lymphocyte 'peptide' culture assay. Reactivity against Melan A/MART-1-derived peptide antigens was readily detectable in both melanoma patients and controls. Reactivity directed against tyrosinase-derived peptide antigens was also detected in both melanoma patients and healthy individuals, but less frequently. A measurable response against gp100/Pmel17-derived antigens was found in 1/10 controls and in 1/26 of the melanoma patients. Reactivity against the influenza matrix peptide was common in both melanoma patients and controls. Our findings show that precursor CTLs against melanocyte differentiation antigens can be detected in peripheral blood of melanoma patients and healthy individuals. The pattern of CTL reactivity directed against melanoma-associated antigens does not seem to be altered in melanoma patients. Despite antigen-specific CTL reactivity, tumour growth was not prevented in melanoma patients and autoimmune phenomena were not detected in healthy individuals. It remains to be determined whether precursor CTLs recognizing melanocyte differentiation antigens can be activated by immunization and lead to effective tumour rejection in vivo.     

Mechanizing some advanced refinement concepts

We describe how the HOL theorem prover can be used to check and apply rules of program refinement. The rules are formulated in the refinement calculus, which is a theory of correctness preserving program transformations. We embed a general command notation with a predicate transformer semantics in the logic of the HOL system. Using this embedding, we express and prove rules for data refinement and superposition refinement of initialized loops. Applications of these proof rules to actual program refinements are checked using the HOL system, with the HOL system generating these conditions. We also indicate how the HOL system is used to prove the verification conditions. Thus, the HOL system can provide a complete mechanized environment for proving program refinements.     

An 8-hour system for Salmonella detection with immunomagnetic separation and homogeneous time-resolved fluorescence PCR

We describe a system consisting of rapid sample enrichment and homogeneous end-point PCR analysis that enables the detection of Salmonella in various food matrices in 8 h. Sample preparation starts with 6 h enrichment step in supplemented broth, after which Salmonella cells are collected with immunomagnetic particles. The particles are washed and dispensed to ready-to-use PCR reaction vessels, which contain dried assay-specific reagents and an internal amplification control. PCR is performed with a novel instrument platform utilising the sensitive label technology of time-resolved fluorometry. Qualitative assay results are automatically interpreted and available in 45 min after sample addition. The overall accuracy, sensitivity and specificity of the Magda CA Salmonella system were 99.1%, 98.4% and 100.0%, respectively, based on the evaluation of 107 samples (beef, pork, poultry and ready-to-eat meals) artificially contaminated with sub-lethally injured Salmonella cells.     

Anaerobic on-site treatment of black water and dairy parlour wastewater in UASB-septic tanks at low temperatures

Anaerobic on-site treatment of synthetic black water (BW) and dairy parlour wastewater (DPWW) was studied in two-phased upflow anaerobic sludge blanket (UASB)-septic tanks at low temperatures (10-20 degrees C). At all temperatures, total chemical oxygen demand (COD(t)) removal was above 90% with BW and above 80% with DPWW and removal of total suspended solids (TSS) above 90% with both wastewaters. Moreover, dissolved COD (COD(dis)) removal was approx. 70% with both wastewaters indicating good biological activity of the sludges. With BW, a single-phased reactor was found sufficient for good COD removals, while with DPWW, a two-phased process was required. Temperature optimum of reactor sludges was still 35 degrees C after long (398d) operation. Most of the nutrients from BW were removed with TSS, while with DPWW nutrient removal was low. In conclusion, UASB-septic tank was found feasible for (pre)treatment of BW and DPWW at low temperatures.     

Alkylresorcinols as antioxidants: hydrogen donation and peroxyl radical‐scavenging effects

Alkylresorcinols, resorcinol derivatives with a long hydrocarbon chain at position 5 of the phenolic ring, have been reported to act as antioxidants and protect polyunsaturated fatty acids against peroxidation. Their structures with meta‐dihydroxy substitutions do not, however, support active hydrogen donation and scavenging of peroxyl radicals responsible for propagation of lipid oxidation. This study compares the antioxidant properties of 5‐n‐pentadecylresorcinol and α‐tocopherol using two model systems: (i) the DPPH· model to test their ability for hydrogen atom donation; and (ii) a triacylglycerol oxidation model to test their ability for scavenging peroxyl radicals. In agreement with theoretical expectations, results from both models showed 5‐n‐pentadecylresorcinol to have very low hydrogen donation and peroxyl radical‐scavenging potency. © 2000 Society of Chemical Industry     

Transient Receptor Potential Ankyrin 1 (TRPA1)—An Inflammation-Induced Factor in Human HaCaT Keratinocytes

Transient receptor potential ankyrin 1 (TRPA1) is an ion channel mainly studied in sensory neurons where it mediates itch, pain and neurogenic inflammation. Recently, some nonneuronal cells have also been shown to express TRPA1 to support inflammatory responses. To address the role of TRPA1 in skin inflammation, we aimed to investigate TRPA1 expression in keratinocytes. HaCaT cells (a model of human keratinocytes) and skin biopses from wild-type and TRPA1 deficient mice were used in the studies. TRPA1 expression in nonstimulated keratinocytes was very low but significantly inducible by the proinflammatory cytokine tumor necrosis factor (TNF) in an nuclear factor kappa B (NF-κB), and mitogen-activated protein (MAP) kinase (p38 and c-Jun N-terminal kinase, JNK)-dependent manner. Interestingly, drugs widely used to treat skin inflammation, the calcineurin inhibitors tacrolimus and cyclosporine and the glucocorticoid dexamethasone, significantly decreased TRPA1 expression. Furthermore, pharmacological inhibition and genetic deletion of TRPA1 reduced the synthesis of TNF-induced monocyte chemoattractant protein 1 (MCP-1) in keratinocytes and mouse skin biopsies. In conclusion, these findings point to an inflammatory role for TRPA1 in keratinocytes and present TRPA1 as a potential drug target in inflammatory skin diseases.