Lifestyle factors and comorbidities in gout patients compared to the general population in Western Sweden: results from a questionnaire study.

OBJECTIVE: This study aimed to identify lifestyle factors associated with gout in patients with prevalent gout compared to the general population. METHOD: Adult patients with gout identified in primary and secondary care in Western Sweden between 2015 and 2017 were sent a questionnaire asking about demographics, lifestyle, and comorbidities. Five age- and gender-matched controls were identified in a random sample of 52 348 individuals aged 16-84 years who participated in the National Public Health survey in Sweden, year 2015. Logistic regression models were used to compare cases and controls with regard to lifestyle factors and comorbidities. RESULTS: Of the 1589 invited gout patients, 868 (55%) responded. After matching for age and gender, 728 were included in the analysis (82.4% male; mean ± sd age 69.3 ± 10.5 years for men and 71.8 ± 9.9 years for women with gout). Male and female gout patients were significantly more likely to be overweight or obese (men 79% vs 66%; women 78.5% vs 65.3%), to have binge-drinking behaviour (men 29.9% vs 11%; women 13.7% vs 2.9%), and to be ex-smokers, compared to controls. Moreover, male gout patients reported lower levels of physical activity, while diabetes and hypertension were more common in both genders with gout than in controls. CONCLUSION: In this questionnaire study, gout patients reported significantly more obesity and binge-drinking behaviour and less physical activity than controls. This suggests that there are great unmet needs for the management of lifestyle factors, particularly regarding overweight/obesity and binge drinking, in patients with gout.

HIV reservoirs and the possibility of a cure for HIV infection.

Recent studies demonstrate that suppressive therapy can drive HIV-1 RNA levels to less than 50 copies mL(-1) in patient plasma. Yet, ultrasensitive assays show that most patients continue to harbour low-level persistent viremia. Treatment intensification studies indicate that low-level viremia could arise from several different sources. These sources include: (i) long-lived HIV-infected cells that replicate and produce virus; (ii) ongoing replication cycles in cells located in sanctuary sites where drug levels are suboptimal; and/or (iii) proliferation of latently infected cells with regeneration of a stable reservoir of slowly dividing infected cells. A well-defined latent reservoir of HIV is memory CD4+ T-cells where latency is established when an activated CD4+ T-cell becomes infected by HIV, but transitions to a terminally differentiated memory cell before it is eliminated. This review examines the dynamics and possible reservoirs of persistent HIV in patients on suppressive therapy, the mechanisms promoting viral latency and strategies to purge latent viral reservoirs. The promising research described here takes a number of steps forward to seriously address HIV remission and/or eradication.

Evidence for kinship between diverse G-protein coupled receptors.

In an earlier publication we described similarities at the primary sequence level between the first probable plant G-protein coupled receptor (GPCR) and three GPCR families (families A, B and F according to Kolakowski's classification) that were previously considered evolutionarily unrelated. Here we analyze further the relatedness among different GPCR families. By using PSI-BLAST, which is a search algorithm that is more potent in detecting weak similarities, one finds additional similarities between GPCR families that have not previously been described. Based on these comparisons, it is possible to divide all the GPCR families into one large clade and two smaller ones. The large clade includes the rhodopsin family (family A), the glucagon receptor family (family B), cyclic AMP receptors (family F), an Arabidopsis thaliana receptor, the Frizzled family and probably also the STE3 pheromone receptors (family E) and vomeronasal receptors type 1. The smaller clades consist of, in one case, BOSS and the GABA-B family of receptors (family C), and in the other the STE2 pheromone receptors (family D) alone. Although our findings are likely to reflect a common ancestry within each of these clades, whether or not two or all three of the clades also share an even more ancient ancestor between them remains an open question that cannot be answered from our present data.

Cellular signalling of PCH-induced pigment aggregation in the crustacean Macrobrachium potiuna erythrophores.

The cellular system responsible for the transduction of the pigment-concentrating hormone (PCH) signal was investigated in erythrophores of the freshwater shrimp, Macrobrachium potiuna. Dose-response curves to the hormone were determined in the absence and in the presence of several drugs that affect sequential steps of the Ca(2+)-dependent signalling pathway. Additionally, the ability of forskolin to induce pigment dispersion was evaluated. Neomycin sulphate (10(-4) and 10(-3) mol.l-1), trifluoperazine (10(-5) and 10(-4) mol.l-1), 1-(5-isoquinolinesulfonlyl)-2-methylpiperazine (10(-7) and 10(-5) mol.l-1) and okadaic acid (10(-7) mol.l-1) significantly (P < 0.05) decreased the responses to PCH. However, okadaic acid at low concentration (10(-9) mol.l-1) and cyclosporin A (10(-6) and 10(-5) mol.l-1) did not significantly (P > 0.05) affect PCH activity. Forskolin (10(-4) mol.l-1) was able to half-maximally reverse the hormone-induced aggregation. Our results suggest that the pigment-concentrating hormone induces pigment aggregation through a Ca(2+)-dependent pathway with a posteriori phosphatase activation, probably the serine/threonine phosphatase 1.

Cloning of a putative G-protein-coupled receptor from Arabidopsis thaliana.

We have cloned and characterized a cDNA from Arabidopsis thaliana that most likely encodes a novel member of the vast superfamily of G-protein-coupled receptor proteins (GPCRs). By taking advantage of amino acid sequence similarities between plant expressed sequence tags (ESTs) and established G-protein-coupled receptor sequences, a probe was obtained which was used for the screening of an Arabidopsis cDNA library. The cDNA which was found is very infrequently represented in the cDNA library, suggesting a low and/or spatially restricted expression. A region of the translated sequence of the cDNA shows the highest similarity to cAMP receptors from the slime mold Dictyostelium discoideum. The same region is also similar to that in members of the animal calcitonin family of receptors. Another region of the putative receptor, however, is similar to sequences of serotonin receptors and other receptors of the so-called rhodopsin family of GPCRs. The rhodopsin family has numerous members in higher vertebrate species. Alignments and phylogenetic analyses of the regions of similarity yielded results in accordance with other evolutionary considerations. Our cDNA thus occurred on a distinct major branch in relation to the rest of the rhodopsin family. In relation to the calcitonin family, our cDNA and cAMP receptors occurred together on a distinct major branch but appear to have diverged from each other shortly after their divergence from the rest of the calcitonin family. Other features further argue for a tentative identification of it as a GPCR. It displays seven discrete and strongly predicted transmembrane domains when analyzed in hydropathy plots. The preferred orientation is with the amino terminus towards the outside. It has one Cys residue in extracellular loop 1 and another in extracellular loop 2. Cys residues in these loops are known to form disulfide bridges in many other GPCRs. Finally, it has several fully conserved amino acids that belong to the most conserved in previously known GPCRs, that occur in the above regions of similarity.

Cytoskeleton and PCH-induced pigment aggregation in Macrobrachium potiuna erythrophores.

Herein we report the effects of microtubule- and actin-like filament disrupting drugs, as well as the microtubule stabilizer taxol, on PCH-induced pigment granule aggregation within erythrophores of the freshwater crustacean Macrobrachium potiuna. Dose-response curves (DRCs) to the pigment-concentrating hormone PCH were determined under control and experimental conditions to evaluate the effects elicited by the cytoskeleton-affecting drugs. Colchicine, at temperatures 22 degrees C and 4 degrees C, and vinblastine significantly inhibited the aggregating response to PCH and affected the dynamics of the process, as shown by the change in the slope of the regression curve calculated from the DRCs. Lumicolchicine, a colchicine analogue with no affinity for tubulin, also inhibited pigment migration, though no change in the slope of the regression curve was observed. The inhibitory effects of lumicolchicine demonstrate that changes in sites other than cytoskeleton, such as membrane permeability, may also cause a decrease in the PCH-induced aggregating responses and that the colchicine effects may result from its action on cellular sites additional to the cytoskeleton. Taxol, a microtubule stabilizer, did not affect the DRC to PCH, and DMSO improved the PCH-evoked responses, pointing out to the maintenance of tubulin in the polymerized state as the appropriate condition for aggregation. Cytochalasin B, an actin-like filament disrupter, diminished the aggregating responses to the hormone, with no change in the slope of the regression curve, indicating that these elements take part in the process and that cytosolic calcium rise, sol/gel transformations and endoplasmic reticulum motility may well play an important role in granule migration. It is suggested that microtubules are steadily polymerized as a requirement for pigment aggregation and that process is biphasic, the initial phase being dependent on the microtubule integrity.

Deletion analysis of a 2S seed storage protein promoter of Brassica napus in transgenic tobacco.

The promoter and upstream region of the Brassica napus 2S storage protein napA gene were studied to identify cis-acting sequences involved in developmental seed-specific expression. Fragments generated by successive deletions of the 5' control region of the napA gene were fused to the reporter gene beta-glucuronidase (GUS). These constructs were used to transform tobacco leaf discs. Analyses of GUS activities in mature seeds from the transformed plants indicated that there were both negatively and positively acting sequences in the napin gene promoter. Deletion of sequences between -1101 and -309 resulted in increased GUS activity. In contrast, deletion of sequences between -309 and -211 decreased the expression. The minimum sequence required for seed-specific expression was a 196 bp fragment between -152 and +44. Further 5' deletion of the fragment to -126 abolished this activity. Sequence comparison showed that a G box-like sequence and two sequence motifs conserved between 2S storage protein genes are located between -148 to -120. Histochemical and fluorometric analysis of tobacco seeds showed that the spatial and developmental expression pattern was retained in the deletion fragments down to -152. However, the expression in tobacco seeds differed from the spatial and temporal expression in B. napus. In tobacco, the napA promoter directed GUS activity early in the endosperm before any visible activity could be seen in the heart-shaped embryo. Later, during the transition from heart to torpedo stages, the main expression of GUS was localized to the embryo. No significant GUS activity was found in either root or leaf.

Characterization of a Brassica napus gene encoding a cruciferin subunit: estimation of sizes of cruciferin gene families.

A gene encoding a subunit of the 12S storage globulin, cruciferin, in Brassica napus (oilseed rape) has been isolated and characterized. The gene consists of about 2200 bp including three short intervening sequences. Primer extension analysis showed that the major transcription start site is located 30 bp 5' of the predicted ATG start codon. This gene belongs to one of three different major families encoding cruciferin subunits. By use of gene-family-specific probes and Southern blotting analysis the number of genes of the three different cruciferin subtypes in B. napus was estimated.

Cloning of a cDNA for rape chloroplast 3-isopropylmalate dehydrogenase by genetic complementation in yeast.

Both insect and mammalian genes have previously been cloned by genetic complementation in yeast. In the present report, we show that the method can be applied also to plants. Thus, we have cloned a rape cDNA for 3-isopropylmalate dehydrogenase (IMDH) by complementation of a yeast leu2 mutation. The cDNA encodes a 52 kDA protein which has a putative chloroplast transit peptide. The in vitro made protein is imported into chloroplasts, concomitantly with a proteolytic cleavage. We conclude that the rape cDNA encodes a chloroplast IMDH. However, Southern analysis revealed that the corresponding gene is nuclear. In a comparison of IMDH sequences from various species, we found that the rape IMDH is more similar to bacterial than to eukaryotic proteins. This suggests that the rape gene could be of chloroplast origin, but has moved to the nucleus during evolution.

Analysis of the promoter region of napin genes from Brassica napus demonstrates binding of nuclear protein in vitro to a conserved sequence motif.

Napin is a seed storage protein from Brassica napus (rape) that is encoded by a gene family. We have isolated and characterized a novel napin gene, napB. Comparisons of the 5'-upstream region of napB to the promoter regions of previously published napin genes reveal that certain sequence motives are evolutionary conserved and may be implicated in gene regulation. These consensus motives, that overlap with purine/pyrimidine stretches, are TACACAT and CATGCA both of which frequently occur as overlapping, direct repeats. Related or identical sequences are also found in the upstream regions of the homologous genes of Arabidopsis thaliana. One copy of the CATGCA motif occurs in close proximity to the TATA box in all the above genes. In this case it overlaps with an octamer sequence (ATGCAAAT) which is a sequence element common in many eukaryotic promoters and enhancers. The TACACAT sequence, as part of a longer purine/pyrimidine stretch, was found to interact with a protein present in crude nuclear extracts from developing B. napus seeds. Napin genes appear to be methylated to almost equal extents whether present in expressing or non-expressing tissue.

Immunological characterization of rapeseed myrosinase.

A purified 75-kDa myrosinase and a crude rapeseed myrosinase fraction were used as antigens to produce mouse anti-myrosinase monoclonal antibodies. The 75-kDa myrosinase was also used to produce a polyclonal rabbit antiserum. The antiserum and one monoclonal antibody reacted with three distinct rapeseed polypeptides of 75, 70 and 65 kDa (M75, M70 and M65, respectively). A second set of monoclonal antibodies reacted exclusively with the 75-kDa form of myrosinase, and a third set showed specificity towards two components of 52 and 50 kDa (myrosinase-binding proteins, MBP52 and MBP50, respectively). MBP52 and MBP50 lack inherent myrosinase activity, but are nevertheless capable of mediating immunoprecipitation of myrosinase due to their interaction with myrosinase. Gel chromatography and glycerol gradient centrifugation experiments resolved two myrosinase-containing fractions. One of these had an approximate molecular mass of 140 kDa and consisted of disulfide-linked dimers of the 75-kDa myrosinase. The other fraction was heterogeneous in size with molecular masses ranging from 250 kDa to approximately 1 MDa. The high-molecular-mass fractions contained complexes consisting of disulfide-linked 70-kDa and 65-kDa myrosinases and non-covalently bound 52-kDa and 50-kDa myrosinase-binding proteins.

Characterization of a cDNA clone encoding a Brassica napus 12 S protein (cruciferin) subunit. Relationship between precursors and mature chains.

Cruciferin (12 S globulin) is a large, neutral, oligometric protein synthesized in rapeseed (Brassica napus) during seed development. It is the major seed protein and is composed of six subunit pairs. Each of these pairs is synthesized as a precursor containing one heavy alpha-chain and one light beta-chain. Electrophoretic analysis of cruciferin showed that four different alpha- and four different beta-chains exist. A cruciferin clone was selected from an embryo cDNA library. This clone, pCRU1, contains a 1518-base pair open reading frame corresponding to a truncated NH2-terminal signal sequence followed by an alpha-chain of 296 and a beta-chain of 190 amino acid residues. Individual cruciferin chains as well as peptides thereof were subjected to NH2-terminal amino acid sequence analysis. The sequences obtained from a specific alpha- and beta-chain pair (alpha 1 and beta 1) showed total identity with the deduced amino acid sequence from pCRU1. Further comparisons revealed that a previously characterized cruciferin cDNA clone encodes one of the precursors for the closely related alpha 2/ alpha 3-beta 2/beta 3 subunits. The deduced amino acid sequences of the two cDNA clones display 64% similarity.

Quantitative in vitro assay for crustacean chromatophorotropins and other pigment cell agonists.

An in vitro crustacean (freshwater shrimp, Macrobrachium potiuna) erythrophore bioassay for chromatophorotropins and other pigment cell agonists is described. The present assay is a quantitative method that determines the pigment responses with the aid of an ocular micrometer. The pigment granules within the erythrophores are dispersed out into the dendritic processes of the cells when the isolated carapace is placed in physiological solution. This bioassay provides, therefore, a method for measuring the response of the pigment cells to aggregating agents such as pigment concentrating hormone (PCH). This bioassay is sensitive to PCH at a concentration as low as 3 x 10(-12) M. Calcium ionophore A23187 mimics the actions of PCH, but, unlike the hormone, the ionophore-induced pigment aggregation is irreversible after physiological solution rinses. Therefore, chromatophorotropic activities of pigment dispersing agents, such as pigment dispersing hormones (PDH), can be determined on ionophore-treated erythrophores. The potencies of alpha-PDH and beta-PDH show a threefold difference (not significant). Because of its convenience and its ability to make an objective determination of the bidirectional pigment movements within erythrophores, this bioassay is a suitable method for further structure-activity studies of the various chromatophorotropins and their analogs.

Guidelines (1988) for training in clinical laboratory management. International Federation of Clinical Chemistry (IFCC) Education Division and International Union of Pure and Applied Chemistry (IUPAC) Clinical Chemistry Division Commission on Teaching of Clinical Chemistry.

Trainees in laboratory medicine must develop skills in laboratory management. Guidelines are detailed for laboratory staff in training, directors responsible for staff development and professional bodies wishing to generate material appropriate to their needs. The syllabus delineates the knowledge base required and includes laboratory planning and organisation, control of operations, methodology and instrumentation, data management and statistics, financial management, clinical use of tests, communication, personnel management and training, and research and development. Methods for achievement of the skills required are suggested. A bibliography of IFCC publications and other material is provided to assist in training in laboratory management.

International Federation of Clinical Chemistry (IFCC), Education Division. Guidelines (1988) for training in clinical laboratory management.

Trainees in laboratory medicine must develop skills in laboratory management. Guidelines are detailed for laboratory staff in training, directors responsible for staff development and professional bodies wishing to generate material appropriate to their needs. The syllabus delineates the knowledge base required and includes laboratory planning and organisation, control of operations, methodology and instrumentation, data management and statistics, financial management, clinical use of tests, communication, personnel management and training, and research and development. Methods for achievement of the skills required are suggested. A bibliography of IFCC publications and other material is provided to assist in training in laboratory management.

International Federation of Clinical Chemistry Education Division and International Union of Pure and Applied Chemistry Clinical Chemistry Division Commission on Teaching of Clinical Chemistry. Guidelines (1988) for training in clinical laboratory management.

Trainees in laboratory medicine must develop skills in laboratory management. Guidelines are detailed for laboratory staff in training, directors responsible for staff development and professional bodies wishing to generate material appropriate to their needs. The syllabus delineates the knowledge base required and includes laboratory planning and organisation, control of operations, methodology and instrumentation, data management and statistics, financial management, clinical use of tests, communication, personnel management and training, and research and development. Methods for achievement of the skills required are suggested. A bibliography of IFCC publications and other material is provided to assist in training in laboratory management.

Guidelines (1988) for training in clinical laboratory management.

Trainees in laboratory medicine must develop skills in laboratory management. Guidelines are detailed for laboratory staff in training, directors responsible for staff development and professional bodies wishing to generate material appropriate to their needs. The syllabus delineates the knowledge base required and includes laboratory planning and organization, control of operations, methodology and instrumentation, data management and statistics, financial management, clinical use of tests, communication, personnel management and training and research and development. Methods for achievement of the skills required are suggested. A bibliography of IFCC publications and other material is provided to assist in training in laboratory management.