Vitamin D in Australia : issues and recommendations

BACKGROUND A significant number of Australians and people from specific groups within the community are suffering from vitamin D deficiency. It is no longer acceptable to assume that all people in Australia receive adequate vitamin D from casual exposure to sunlight.OBJECTIVE This article provides information on causes, consequences, treatment and prevention of vitamin D deficiency in Australia. DISCUSSION People at high risk of vitamin D deficiency include the elderly, those with skin conditions where avoidance of sunlight is required, dark skinned people (particularly women during pregnancy or if veiled) and patients with malabsorption, eg. coeliac disease. For most people, deficiency can be prevented by 5&ndash;15 minutes exposure of face and upper limbs to sunlight 4&ndash;6 times per week. If this is not possible then a vitamin D supplement of at least 400 IU* per day is recommended. In cases of established vitamin D deficiency, supplementation with 3000-5000 IU per day for at least 1 month is required to replete body stores. Increased availability of larger dose preparations of cholecalciferol would be a useful therapy in the case of severe deficiencies. * 40 IU (international units) = 1 &micro;g<br /

Cold Induction of EARLI1, a Putative Arabidopsis Lipid Transfer Protein, Is Light and Calcium Dependent

As sessile organisms, plants must adapt to their environment. One approach toward understanding this adaptation is to investigate environmental regulation of gene expression. Our focus is on the environmental regulation of EARLI1, which is activated by cold and long-day photoperiods. Cold activation of EARLI1 in short-day photoperiods is slow, requiring several hours at 4ºC to detect an increase in mRNA abundance. EARLI1 is not efficiently cold-activated in etiolated seedlings, suggesting that photomorphogenesis is necessary for its cold activation. Cold activation of EARLI1 is inhibited in the presence of the calcium channel blocker lanthanum chloride or the calcium chelator EGTA. Addition of the calcium ionophore Bay K8644 results in cold-independent activation of EARLI1. These data suggest that EARLI1 is not an immediate target of the cold response, and that calcium flux affects its expression. EARLI1 is a putative secreted protein and has motifs found in lipid transfer proteins. Over-expression of EARLI1 in transgenic plants results in reduced electrolyte leakage during freezing damage, suggesting that EARLI1 may affect membrane or cell wall stability in response to low temperature stress

The Role of Voltage-Gated Calcium Channels in Neurotransmitter Phenotype Specification: Coexpression and Functional Analysis in Xenopus laevis

Calcium activity has been implicated in many neurodevelopmental events, including the specification of neurotransmitter phenotypes. Higher levels of calcium activity lead to an increased number of inhibitory neural phenotypes, whereas lower levels of calcium activity lead to excitatory neural phenotypes. Voltage-gated calcium channels (VGCCs) allow for rapid calcium entry and are expressed during early neural stages, making them likely regulators of activity-dependent neurotransmitter phenotype specification. To test this hypothesis, multiplex fluorescent in situ hybridization was used to characterize the coexpression of eight VGCC alpha 1 subunits with the excitatory and inhibitory neural markers xVGlut1 and xVIAAT in Xenopus laevis embryos. VGCC coexpression was higher with xVGlut1 than xVIAAT, especially in the hindbrain, spinal cord, and cranial nerves. Calcium activity was also analyzed on a single-cell level, and spike frequency was correlated with the expression of VGCC alpha 1 subunits in cell culture. Cells expressing Ca(V)2.1 and Ca(V)2.2 displayed increased calcium spiking compared with cells not expressing this marker. The VGCC antagonist diltiazem and agonist (-)BayK 8644 were used to manipulate calcium activity. Diltiazem exposure increased the number of glutamatergic cells and decreased the number of gamma-aminobutyric acid (GABA)ergic cells, whereas (2) BayK 8644 exposure decreased the number of glutamatergic cells without having an effect on the number of GABAergic cells. Given that the expression and functional manipulation of VGCCs are correlated with neurotransmitter phenotype in some, but not all, experiments, VGCCs likely act in combination with a variety of other signaling factors to determine neuronal phenotype specification. (C) 2014 Wiley Periodicals, Inc

Role of glucose and CcpA in capsule expression and virulence of Streptococcus suis

Streptococcus suis is one of the most important pathogens in pigs and is also an emerging zoonotic agent. After crossing the epithelial barrier, S. suis causes bacteraemia, resulting in meningitis, endocarditis and bronchopneumonia. Since the host environment seems to be an important regulatory component for virulence, we related expression of virulence determinants of S. suis to glucose availability during growth and to the sugar metabolism regulator catabolite control protein A (CcpA). We found that expression of the virulence-associated genes arcB, representing arcABC operon expression, cps2A, representing capsular locus expression, as well as sly, ofs, sao and epf, differed significantly between exponential and early stationary growth of a highly virulent serotype 2 strain. Deletion of ccpA altered the expression of the surface-associated virulence factors arcB, sao and eno, as well as the two currently proven virulence factors in pigs, ofs and cps2A, in early exponential growth. Global expression analysis using a cDNA expression array revealed 259 differentially expressed genes in early exponential growth, of which 141 were more highly expressed in the CcpA mutant strain 10¿ccpA and 118 were expressed to a lower extent. Interestingly, among the latter genes, 18 could be related to capsule and cell wall synthesis. Correspondingly, electron microscopy characterization of strain 10¿ccpA revealed a markedly reduced thickness of the capsule. This phenotype correlated with enhanced binding to porcine plasma proteins and a reduced resistance to killing by porcine neutrophils. Taken together, our data demonstrate that CcpA has a significant effect on the capsule synthesis and virulence properties of S. suis

Comparison of Fit Between Zirconia and Metal Copings Fabricated Conventionally or Using Different CAD/CAM Technique

This study was designed to assess the fit of cobalt chromium copings fabricated using direct metal laser sintering and to compare with copings fabricated from nickel chromium (traditional casting) and zirconia (milled from partially sintered blanks). For both cobalt chromium and zirconia groups, impressions were generated using direct or indirect digitization. There were 5 experimental groups of 12 copings fabricated for a prepared maxillary right molar. The intimacy of fit (measured at the margin, axial wall and occlusal) was assessed using a replica method technique. The results showed that zirconia copings produced using direct digitization had significantly smaller marginal, axial and occlusal measurements compared to other groups (p⟨0.05). Nevertheless, all groups had a mean marginal gap within accepted clinical values. Though there was a decrease in the intimacy of fit from the margin to axial wall to the occlusal surface. It can be concluded that the fit of zirconia copings fabricated using direct digitization was significantly better than the other groups. However, it can be expected that further enhancement and refinement of additive technology such as direct metal laser sintering will offer clinicians a viable alternative to nickel chromium analogue production methods in the future

Universal solvent quality crossover of the zero shear rate viscosity of semidilute DNA solutions

The scaling behaviour of the zero shear rate viscosity of semidilute unentangled DNA solutions, in the double crossover regime driven by temperature and concentration, is mapped out by systematic experiments. The viscosity is shown to have a power law dependence on the scaled concentration $c/c^*$, with an effective exponent that depends on the solvent quality parameter $z$. The determination of the form of this universal crossover scaling function requires the estimation of the $\theta$ temperature of dilute DNA solutions in the presence of excess salt, and the determination of the solvent quality parameter at any given molecular weight and temperature. The $\theta$ temperature is determined to be $T_\theta \approx 15^\circ$ C using static light scattering, and the solvent quality parameter has been determined by dynamic light scattering.Comment: 39 pages, 26 figures, accepted in Journal of Rheology. Includes supplemental material

Modelling flow through a permeable bed: a combined physical-numerical approach

River hydrodynamicsTurbulent open channel flow and transport phenomen

Fine-scale time-lapse analysis of the biphasic, dynamic behaviour of the two Vibrio cholerae chromosomes

Using fluorescent repressor-operator systems in live cells, we investigated the dynamic behaviour of chromosomal origins in Vibrio cholerae, whose genome is divided between two chromosomes. We have developed a method of analysing fine-scale motion in the curved co-ordinate system of vibrioid bacteria. Using this method, we characterized two different modes of chromosome behaviour corresponding to periods between segregation events and periods of segregation. Between segregation events, the origin positions are not fixed but rather maintained within ellipsoidal caged domains, similar to eukaryotic interphase chromosome territories. These domains are approximately 0.4 µm wide and 0.6 µm long, reflecting greater restriction in the short axis of the cell. During segregation, movement is directionally biased, speed is comparable between origins, and cell growth can account for nearly 20% of the motion observed. Furthermore, the home domain of each origin is positioned by a different mechanism. Specifically, the oriCI domain is maintained at a constant actual distance from the pole regardless of cell length, while the oriCII domain is maintained at a constant relative position. Thus the actual position of oriCII varies with cell length. While the gross behaviours of the two origins are distinct, their fine-scale dynamics are remarkably similar, indicating that both experience similar microenvironments

Purification and Reconstitution of the Glutamate Carrier GltT of the Thermophilic Bacterium Bacillus stearothermophilus

An affinity tag consisting of six adjacent histidine residues followed by an enterokinase cleavage site was genetically engineered at the N-terminus of the glutamate transport protein GltT of the thermophilic bacterium Bacillus stearothermophilus. The fusion protein was expressed in Escherichia coli and shown to transport glutamate. The highest levels of expression were observed in E. coli strain DH5α grown on rich medium. The protein could be purified in a single step by Ni2+-NTA affinity chromatography after solubilization of the cytoplasmic membranes with the detergent Triton X100. Purified GltT was reconstituted in an active state in liposomes prepared from E. coli phospholipids. The protein was reconstituted in detergent-treated preformed liposomes, followed by removal of the detergent with polystyrene beads. Active reconstitution was realized with a wide range of Triton X100 concentrations. Neither the presence of glycerol, phospholipids, nor substrates of the transporter was necessary during the purification and reconstitution procedure to keep the enzyme in an active state. In B. stearothermophilus, GltT translocates glutamate in symport with protons or sodium ions. In membrane vesicles derived from E. coli cells expressing GltT, the Na+ ion dependency seems to be lost, suggesting a role for the lipid environment in the cation specificity. In agreement with the last observation, glutamate transport catalyzed by purified GltT reconstituted in E. coli phospholipid is driven by an electrochemical gradient of H+ but not of Na+.