Capsular arabinomannans from Mycobacterium avium with morphotype-specific structural differences but identical biological activity

The capsules of two colony morphotypes of Mycobacterium avium strain 2151 were investigated, i.e. the virulent smooth-transparent (SmT1) and the nonvirulent smooth-opaque (SmO) types. From both morphotypes we separated a nonacylated arabinomannan (AM) from an acylated polysaccharide fraction by affinity chromatography, of which the AMs were structurally characterized. The AMs from the virulent morphotype, in contrast to that from the nonvirulent form, possessed a larger mannan chain and a shorter arabinan chain. Incubation of murine bone marrow-derived macrophages and human dendritic cells showed that the acylated polysaccharide fractions were potent inducers of tumor necrosis factor-alpha, interleukin-12, and interleukin-10 compared with nonacylated AMs, which led to only a marginal cytokine release. Further in vitro experiments showed that both the acylated polysaccharide fractions and the nonacylated AMs were able to induce in vitro anti-tumor cytotoxicity of human peripheral blood mononuclear cells. Thus, morphotype-specific structural differences in the capsular AMs of M. avium do not correlate with biological activity; however, their acylation is a prerequisite for effective stimulation of murine macrophages and human dendritic cells.     

New insights into the biosynthesis of mycobacterial lipomannan arising from deletion of a conserved gene

Genetic construction of a mutant strain (designated MSMEG4245) of Mycobacterium smegmatis, defective in a broadly conserved gene for a putative glycosyltransferase of the glycosyltransferase-C superfamily, results in a phenotype marked by the virtual absence of the phosphatidylinositol-containing lipomannan and lipoarabinomannan, replaced instead by a novel truncated form of lipomannan. The normal spectrum of phosphatidylinositol mannosides, long presumed precursors of these lipoglycans, was retained. Matrix-assisted laser desorption/ionization-time of flight/mass spectrometry of the mutated form of lipomannan shows a family of phosphatidylinositol-anchored lipomannans with from only 5 to 20 Manp residues as compared with lipomannan from the wild type strain consisting of 21-34 Manp residues but with few changes in the branching pattern. Thus, MSMEG4245 is apparently a key mannosyltransferase, required for the proper elongation of lipomannan to its normal state and subsequent synthesis of lipoarabinomannan. The corresponding ortholog in Mycobacterium tuberculosis H37Rv has been identified as Rv2174. This previously unrecognized feature of the biosynthesis of lipomannan/lipoarabinomannan allows a significant revision of structural and biosynthetic schemata and provides a molecular basis of selectivity in biosynthesis, as conferred by the MSMEG4245 gene.     

A small change approach to prevent long-term weight gain in adults with overweight and obesity: a randomized controlled trial

Background:Efforts to manage obesity through weight loss are often unsuccessful as most adults are not able to sustain the major changes in behaviour that are required to maintain weight loss long term. We sought to determine whether small changes in physical activity and diet prevent weight gain in adults with overweight and obesity.Methods:We randomized 320 sedentary adults with overweight or obesity to monitoring alone (MA, n = 160) or a small change approach (SCA, n = 160). In Phase I (2 yr), MA participants were asked to maintain their normal lifestyle and SCA participants were counselled to make small changes in diet and physical activity, namely a suggested increase in daily step count of 2000 steps with a decrease in energy intake of 100 kcal per day, with group and individual support. Phase II (1 yr) was a passive follow-up period. The difference in change in body weight between groups at 24 and 36 months from baseline was the primary outcome. Additional outcomes included waist circumference and cardiorespiratory fitness.Results:Overall, 268 participants (83.8%) completed the 2-year intervention, and 239 (74.7%) returned at the end of the follow-up period at 3 years. The difference in body weight change between the SCA and MA groups was significant at 3, 6, 12 and 15 months from baseline, but was no longer significant at 24 months (mean change 0.9 [standard error (SE) 0.5] kg v. −0.4 [SE 0.5] kg; difference −0.6, 95% confidence interval [CI] −1.9 to 0.8) or at 36 months (−1.2 [SE 0.8] v. −0.7 [SE 0.8] kg; difference −0.5, 95% CI −2.2 to 1.2). Changes in waist circumference and cardiorespiratory fitness were not significantly different between groups at 24 or 36 months (both p > 0.1).Interpretation:The SCA did not prevent weight gain compared with monitoring alone at 2 or 3 years in adults with overweight or obesity. On average, we observed prevention of weight gain in both arms of the trial. Trial registration:ClinicalTrials.gov, no. {"type":"clinical-trial","attrs":{"text":"NCT02027077","term_id":"NCT02027077"}}NCT02027077.

Overweight and obesity contribute to chronic diseases and present a major public health challenge¹ as more than 63% of adults in Canada currently live with overweight or obesity.² Despite the urgent need to address the obesity problem, few strategies designed to reduce obesity have been broadly successful.Results from randomized controlled trials show that most adults are not able to sustain the major changes in behaviour that are required to maintain weight loss long term.^3,4 A more reasonable and achievable goal may be to focus on the prevention of weight gain. Preventing further weight gain is clinically important as even modest weight gain (0.5–1.0 kg/yr) in adults with overweight and obesity has negative associations with adverse outcomes, such as cancer,⁵ all-cause and cardiovascular disease–related death⁶ and poor health-related quality of life.⁷It has been estimated that, for 90% of the adult population, a reduction of about 100–150 kcal/d would be required to prevent positive energy balance.^8,9 Two separate pilot studies have shown that a small change approach (SCA, defined as an increase in daily step count of 2000 steps or a decrease in energy intake of 100 kcal) prevented weight gain in a small group of adults and children with overweight over 13 weeks.^9,10 More recently SCA was shown to be associated with reduced weight gain in young adults with overweight at 2 years, compared with controls.¹¹We sought to determine the effectiveness of the SCA to prevent weight gain in adults with overweight (defined as a body mass index [BMI] 25–29.9) and obesity (defined as a BMI of ≥ 30) at 2 and 3 years. We hypothesized that the SCA would prevent weight gain in comparison with monitoring of body weight alone.     

Extension of Family Planning Service in General Practice

All general practitioners on the list of a single executive council were contacted and 91% were interviewed. They were asked about the type of family planning service currently provided and their willingness to extend the service. Only 3% said that they were not providing any service at the time of interview, but only 4% stated that they provided a complete range of techniques to all patients; almost two thirds of practitioners only provided advice and the “pill”. Lack of training in family planning techniques emerged as the most important factor in determining the type of service provided. At the time of interview 64% of doctors stated that they would like further training, and 35% of all doctors asked for a full course of clinical training. Many general practitioners (81%) were willing to extend their family planning services but 65% wanted financial reimbursement and 50% needed additional administrative support as prerequisites.     

NMR structure of the haem core of a novel tetrahaem cytochrome isolated from Shewanella frigidimarina: identification of the haem-specific axial ligands and order of oxidation

The tetrahaem cytochrome isolated during anaerobic growth of Shewanella frigidimarina NCIMB400 is a small protein (86 residues) involved in electron transfer to Fe(III), which can be used as a terminal respiratory oxidant by this bacterium. A 3D solution structure model of the reduced form of the cytochrome has been determined using NMR data in order to determine the relative orientation of the haems. The haem core architecture of S. frigidimarina tetrahaem cytochrome differs from that found in all small tetrahaem cytochromes c(3) so far isolated from strict anaerobes, but has some similarity to the N-terminal cytochrome domain of flavocytochrome c(3) isolated from the same bacterium. NMR signals obtained for the four haems of S. frigidimarina tetrahaem cytochrome at all stages of oxidation were cross-assigned to the solution structure using the complete network of chemical exchange connectivities. Thus, the order in which each haem in the structure becomes oxidised was determined.     

The Antarctic toothfish (Dissostichus mawsoni) lacks plasma albumin and utilises high density lipoprotein as its major palmitate binding protein

Plasma from the Antarctic toothfish, Dissostichus mawsoni, a member of the advanced teleost Nototheniidae family, was analysed. Agarose gel electrophoresis showed a major diffuse anionic protein that bound [14C]palmitic acid but not 63Ni2+, and two more cationic proteins that bound 63Ni2+ but not palmitate. Oil Red O staining following cellulose acetate electrophoresis indicated that the palmitate binding protein was a lipoprotein. Two-dimensional electrophoresis showed that this palmitate binding band was composed of three proteins with M(r) of 11, 30, and 42 kDa, without any trace of material at approximately 65 kDa, the mass of albumin. N-terminal sequencing of the palmitate binding band gave a major sequence of DAAQPSQELR-, indicating a high degree of homology to apolipoprotein A-I (apo-AI), the major apolipoprotein of high density lipoprotein (HDL). N-terminal sequencing of the major nickel binding band produced a sequence with no homology to albumin. When ultracentrifugation was used to isolate the lipoproteins from Antarctic toothfish plasma, the palmitate binding protein was found solely in the lipoprotein fraction. In competitive binding experiments, added human albumin did not prevent palmitate binding to toothfish HDL. In conclusion, there is no evidence for albumin in Antarctic toothfish plasma and HDL assumes the role of fatty acid transport.     

Zebrafish aussicht mutant embryos exhibit widespread overexpression of ace (fgf8) and coincident defects in CNS development

During the development of the zebrafish nervous system both noi, a zebrafish pax2 homolog, and ace, a zebrafish fgf8 homolog, are required for development of the midbrain and cerebellum. Here we describe a dominant mutation, aussicht (aus), in which the expression of noi and ace is upregulated. In aus mutant embryos, ace is upregulated at many sites in the embryo, while noi expression is only upregulated in regions of the forebrain and midbrain which also express ace. Subsequent to the alterations in noi and ace expression, aus mutants exhibit defects in the differentiation of the forebrain, midbrain and eyes. Within the forebrain, the formation of the anterior and postoptic commissures is delayed and the expression of markers within the pretectal area is reduced. Within the midbrain, En and wnt1 expression is expanded. In heterozygous aus embryos, there is ectopic outgrowth of neural retina in the temporal half of the eyes, whereas in putative homozygous aus embryos, the ventral retina is reduced and the pigmented retinal epithelium is expanded towards the midline. The observation that aus mutant embryos exhibit widespread upregulation of ace raised the possibility that aus might represent an allele of the ace gene itself. However, by crossing carriers for both aus and ace, we were able to generate homozygous ace mutant embryos that also exhibited the aus phenotype. This indicated that aus is not tightly linked to ace and is unlikely to be a mutation directly affecting the ace locus. However, increased Ace activity may underly many aspects of the aus phenotype and we show that the upregulation of noi in the forebrain of aus mutants is partially dependent upon functional Ace activity. Conversely, increased ace expression in the forebrain of aus mutants is not dependent upon functional Noi activity. We conclude that aus represents a mutation involving a locus normally required for the regulation of ace expression during embryogenesis.     

Determination of an Optimal Pitfall Trap Size for Sampling Spiders in a Western Australian Jarrah Forest

Pitfall trapping is a sampling technique extensively used to sample surface foraging invertebrates for biological diversity studies and ecological monitoring. To date, very few invertebrate studies have considered what trap size is optimal for sampling spiders. This study presents preliminary findings from a single short sampling period on the role of trap size in sampling spiders in a Western Australian Jarrah forest. Four different trap diameters (4.3, 7.0, 11.1 and 17.4 cm) were examined (4 trap sizes × 15 replicates = 60 traps). Two-way ANOVAs revealed no significant interaction effects between trap size or the spatial positioning of transects within the study site along which the pitfall traps were arranged. Post-hoc tests revealed abundance, family richness and species richness increased with increasing trap sizes for traps 7.0 cm. No significant differences in these dependent variables occurred between 4.3 and 7.0 cm traps, or for species richness between 11.1 and 17.4 cm traps. Determination of an optimal trap size was undertaken by bootstrapping and calculating species accumulation curves for increasing numbers of traps used. Three different criteria were considered: equivalent number of traps (15), standardized sampling intensity (cumulative trap circumference, approximately 207 cm) and standardized cumulative handling time (approximately 1 hour 17 minutes). The largest trap size (17.4 cm) was most efficient in terms of number of traps and trap circumference. For the same number of traps, it caught 19 species whereas all other trap sizes caught ten species. At the standardized circumference, it caught seven species whereas all other trap sizes caught five. For handling time, however, the two largest trap sizes (17.4 and 11.1 cm) were optimal. Both caught nine species whereas all other traps caught 相似文献