Characterization of scrubber water discharges from ships using comprehensive suspect screening strategies based on GC-APCI-HRMS.

An extended suspect screening approach for the comprehensive chemical characterization of scrubber discharge waters from exhaust gas cleaning systems (EGCSs), used to reduce atmospheric shipping emissions of sulphur oxides, was developed. The suspect screening was based on gas chromatography coupled with high-resolution mass spectrometry (GC-HRMS) and focused on the identification of polycyclic aromatic hydrocarbons (PAHs) and their alkylated derivatives (alkyl-PAHs), which are among the most frequent and potentially toxic organic contaminants detected in these matrices. Although alkyl-PAHs can be even more abundant than parent compounds, information regarding their occurrence in scrubber waters is scarce. For compound identification, an in-house compound database was built, with 26 suspect groups, including 25 parent PAHs and 23 alkyl-PAH homologues. With this approach, 7 PAHs and 12 clusters of alkyl-PAHs were tentatively identified, whose occurrence was finally confirmed by target analysis using GC coupled with tandem mass spectrometry (GC-MS/MS). Finally, a retrospective analysis was performed to identify other relevant (poly)cyclic aromatic compounds (PACs) of potential concern in scrubber waters. According to it, 18 suspect groups were tentatively identified, including biphenyls, dibenzofurans, dibenzothiophenes and oxygenated PAHs derivatives. All these compounds could be used as relevant markers of scrubber water contamination in heavy traffic marine areas and be considered as potential stressors when evaluating scrubber water toxicity.

Mushroom productivity trends in relation to tree growth and climate across different European forest biomes.

Although it is logical to think that mycorrhizal mushroom production should be somehow related to the growth of the trees from which the fungi obtain carbohydrates, little is known about how mushroom yield patterns are related to tree performance. In this study, we delved into the understanding of the relationships between aboveground fungal productivity, tree radial growth patterns and climatic conditions across three latitudinally different bioclimatic regions encompassing Mediterranean, temperate and boreal forest ecosystems in Europe. For this purpose, we used a large assemblage of long-term data of weekly or biweekly mushroom yield monitoring in Spain, Switzerland and Finland. We analysed the relationships between annual mushroom yield (considering both biomass and number of sporocarps per unit area), tree ring features (tree ring, earlywood and latewood widths), and meteorological conditions (i.e. precipitation and temperature of summer and autumn) from different study sites and forest ecosystems, using both standard and partial correlations. Moreover, we fitted predictive models to estimate mushroom yield from mycorrhizal and saprotrophic fungal guilds based on climatic and dendrochronological variables. Significant synchronies between mushroom yield and climatic and dendrochronological variables were mostly found in drier Mediterranean sites, while few or no significant correlations were found in the boreal and temperate regions. We observed positive correlations between latewood growth and mycorrhizal mushroom biomass only in some Mediterranean sites, this relationship being mainly mediated by summer and autumn precipitation. Under more water-limited conditions, both the seasonal wood production and the mushroom yield are more sensitive to precipitation events, resulting in higher synchrony between both variables. This comparative study across diverse European forest biomes and types provides new insights into the relationship between mushroom productivity, tree growth and weather conditions.

pH dependence of the peptide thiol-disulfide oxidase activity of six members of the human protein disulfide isomerase family.

Protein folding in the endoplasmic reticulum is often associated with the formation of native disulfide bonds, a process which in vivo is one of the rate limiting steps of protein folding and which is facilitated by the enzyme protein disulfide isomerase (PDI). Higher eukaryotes have multiple members of the PDI family, for example, seventeen human PDIs have been reported to date. With multiple members of the same family being present, even within the same cell, the question arises as to what differential functions are they performing? To date there has been no systematic evaluation of the enzymological properties of the different members of the PDI-family. To address the question of whether different PDI family members have differing thioldisulfide chemistry, we have recombinantly expressed and purified six members of the family, PDI, PDIp, ERp57, ERp72, P5, and PDIr from a single organism, human. An examination of the pH-dependence and nature of the rate limiting step for the peptide thiol-disulfide oxidase activity of these enzymes reveals that, with the exception of PDIr, they are all remarkably similar. In the light of this data potential differential functions for these enzymes are discussed.

The C/C-13910 genotype of adult-type hypolactasia is associated with an increased risk of colorectal cancer in the Finnish population.

BACKGROUND AND AIMS: The role of nutrition in the pathogenesis of colorectal cancer is not fully understood. Milk products are an essential part of human nutrition in Western countries. Absorption of lactose, the main sugar of milk, is regulated by the activity of the lactase enzyme in the gut wall. The activity of lactase is genetically determined and is associated with a C/T single nucleotide polymorphism residing 13910 bp upstream of the lactase coding sequence. Here we have studied the relationship between the C/T(-13910) polymorphism and colorectal cancer in Finnish, British, and Spanish populations. PATIENTS AND METHODS: A total of 2766 subjects, including 963 Finnish, 283 British, and 163 Spanish subjects with colorectal cancer, and 773 Finnish, 363 British, and 221 Spanish control subjects, were genotyped for the C/T(-13910) variant by polymerase chain reaction minisequencing. RESULTS: The C/C(-13910) genotype, which is a robust molecular marker of low lactase activity (lactase non-persistence), was found to significantly associate with the risk of colorectal cancer (p = 0.015) in the Finnish subjects, with an odds ratio of 1.40 (95% confidence interval 1.07-1.85). No association was found with site, histology, or stage of the tumour. No significant risk was detected in the British or Spanish populations. CONCLUSION: Low lactase enzyme activity, defined by genotyping of the C/T(-13910) variant, may increase the risk of colorectal cancer. Further studies are warranted to investigate the role of milk and other dairy products in the pathogenesis of colon cancer in different populations.

A conserved arginine plays a role in the catalytic cycle of the protein disulphide isomerases.

The pK(a) values of the CXXC active-site cysteine residues play a critical role in determining the physiological function of the thioredoxin superfamily. To act as an efficient thiol-disulphide oxidant the thiolate state of the N-terminal cysteine must be stabilised and the thiolate state of the C-terminal cysteine residue destabilised. While increasing the pK(a) value of the C-terminal cysteine residue promotes oxidation of substrates, it has an inhibitory effect on the reoxidation of the enzyme, which is promoted by the formation of a thiolate at this position. Since reoxidation is essential to complete the catalytic cycle, the differential requirement for a high and a low pK(a) value for the C-terminal cysteine residue for different steps in the reaction presents us with a paradox. Here, we report the identification of a conserved arginine residue, located in the loop between beta5 and alpha4 of the catalytic domains of the human protein disulphide isomerase (PDI) family, which is critical for the catalytic function of PDI, ERp57, ERp72 and P5, specifically for reoxidation. An examination of the published NMR structure for the a domain of PDI combined with molecular dynamic studies suggest that the side-chain of this arginine residue moves into and out of the active-site locale and that this has a very marked effect on the pK(a) value of the active-site cysteine residues. This intra-domain motion resolves the apparent dichotomy of the pK(a) requirements for the C-terminal active-site cysteine.

Defining the domain boundaries of the human protein disulfide isomerases.

The protein disulfide isomerase (PDI) family of folding catalysts are constructed from combinations of redoxactive and redox-inactive domains, all of which are probably based on the thioredoxin fold. To understand the function of each domain in the variety of catalytic reactions that each family member can perform (to differing extents), the domain boundaries of each family member must be known. By using a technique based on sequence alignments and the known structure of the a and b domains of human PDI, we generated a large number of domain constructs for all six redox-active human PDIs: PDI, PDIp, ERp72, ERp57, P5, and PDIr. The ability to generate significant amounts of soluble protein in E. coli from most of these domain constructs strongly indicates that the domain boundaries are correct. The implications for these domain boundaries on the tertiary structure of the human PDIs are discussed.

Basal and acidic fibroblast growth factor-induced atrial natriuretic peptide gene expression and secretion is inhibited by staurosporine.

We examined the mechanisms involved in the activation of atrial natriuretic peptide (ANP) gene expression and secretion in response to acidic fibroblast growth factor (aFGF) by studying the effects of staurosporine, a protein kinase C inhibitor, and 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C, on basal and AFGF-induced ANP messenger RNA (mRNA) and immunoreactive ANP (IR-ANP) levels in cultured neonatal rat cardiac myocytes. Acidic FGF caused a dose- and time-dependent increase in IR-ANP and immunoreactive N-terminal fragment of proANP (IR-NT-proANP) release into the culture medium from ventricular but not from atrial myocytes. In ventricular cells, 50 ng/ml aFGF for 24 or 48 h resulted in a 70% or 181% increase, respectively, in the accumulation of IR-ANP into the culture medium. Acidic FGF also stimulated ANP gene expression significantly; after 48 h of incubation, the ANP mRNA levels of aFGF-treated ventricular myocytes were 205% (P < 0.001) higher than those of control cells. Staurosporine alone at concentration of 10 nM significantly decreased the basal IR-ANP and IR-NT-proANP secretion, and inhibited the aFGF-induced increase in ANP mRNA and IR-ANP levels in ventricular myocytes. TPA (100 nM) alone significantly stimulated ANP gene expression and secretion but these effects were not augmented by combining aFGF with TPA. High performance liquid chromatographical analysis showed that atrial and ventricular myocytes maintained in serum-free medium were capable of secreting processed, ANP99-126 sized material, and that aFGF did not alter the processing of ANP in ventricular cultures. These results demonstrate that aFGF is a potent stimulator of ANP gene expression and secretion in cultured neonatal rat ventricular but not in atrial cells. The observations that (a) staurosporine completely abolished the effects of aFGF on ANP gene expression and release and (b) ANP secretory and gene expression inducing effects of phorbol ester were not augmented by aFGF, suggest an important role of protein kinase C in mediating aFGF-induced ANP gene expression and secretion.

Intraoperative fine needle aspiration cytologic examination of pancreatic lesions.

Despite the possibilities in modern imaging technology and percutaneous biopsy, a surgeon may still find an undiagnosed mass in the pancreas at laparotomy. In this situation, intraoperative fine needle aspiration cytologic (IFNAC) examination has been reported to be helpful. We reviewed our experiences with IFNAC in 98 patients. Fifty patients had a malignant pancreatic tumor as verified on histologic examination. The results of IFNAC correctly suggested a malignant tumor in 35 patients, for an initial sensitivity of 70 per cent. Re-examination of the slides resulted in 81 per cent sensitivity, which was not a significant improvement. The sensitivity rate (an average of 83 per cent in the literature) does not, however, express enough the unreliability of the method in individual patients. We conclude that, although IFNAC correctly differentiates between carcinoma and benign pancreatic diseases in most instances, the justification for pancreas resection cannot always be based on cytologic findings, but rather on clinical and laparotomy findings.

Chondromalacia patellae. Results of operative treatment.

The series consisted of 26 patients operated on in 1972-1978 for chondromalacia patellae. The patients were followed up an average of 4.6 years after the operation. The operation involved one knee in 21 patients, both knees in 5. The primary operation was carried out for lesions of the articular cartilage of the patella alone in 18 cases (in 6 of these a second operation was necessary) and for a biomechanical disturbance of the patellofemoral joint in 13 cases. Degenerative changes of the patellar cartilage were observed at anatomo-pathological examination in 29 of 31 knees. On average, the patients' symptoms were alleviated after the operation, but comparison of different types of operation showed a statistically significant improvement only after operations that modified the biomechanics. The results confirm the view that symptoms originating in the patellofemoral joint often are due to biomechanical disturbances of this joint and the extensor system of the knee, and that the removal of injured cartilage alone is not sufficient.