Hepatitis C Virus Phylogenetic Clustering Is Associated with the Social-Injecting Network in a Cohort of People Who Inject Drugs

It is hypothesized that social networks facilitate transmission of the hepatitis C virus (HCV). We tested for association between HCV phylogeny and reported injecting relationships using longitudinal data from a social network design study. People who inject drugs were recruited from street drug markets in Melbourne, Australia. Interviews and blood tests took place three monthly (during 2005–2008), with participants asked to nominate up to five injecting partners at each interview. The HCV core region of individual isolates was then sequenced and phylogenetic trees were constructed. Genetic clusters were identified using bootstrapping (cut-off: 70%). An adjusted Jaccard similarity coefficient was used to measure the association between the reported injecting relationships and relationships defined by clustering in the phylogenetic analysis (statistical significance assessed using the quadratic assignment procedure). 402 participants consented to participate; 244 HCV infections were observed in 238 individuals. 26 genetic clusters were identified, with 2–7 infections per cluster. Newly acquired infection (AOR = 2.03, 95% CI: 1.04–3.96, p = 0.037, and HCV genotype 3 (vs. genotype 1, AOR = 2.72, 95% CI: 1.48–4.99) were independent predictors of being in a cluster. 54% of participants whose infections were part of a cluster in the phylogenetic analysis reported injecting with at least one other participant in that cluster during the study. Overall, 16% of participants who were infected at study entry and 40% of participants with newly acquired infections had molecular evidence of related infections with at least one injecting partner. Likely transmission clusters identified in phylogenetic analysis correlated with reported injecting relationships (adjusted Jaccard coefficient: 0.300; p<0.001). This is the first study to show that HCV phylogeny is associated with the injecting network, highlighting the importance of the injecting network in HCV transmission.     

Intracellular pH of frog sartorius muscle

A weak base, morpholine, has been labelled with ³H and tested for its suitability as an indicator for intracellular pH, by distribution in the tissue water of frog sartorius muscle in the species Hyla litoria. Its pK'a at 20°C in a solution of the same of ionic strength as frog Ringer was found to be 8.45 ± 0.02, which is in the range of maximal sensitivity. Morpholine equilibrated with the tissue in 17 h; it was shown that it was not bound to intracellular constituents, that it was not metabolised nor toxic in the concentrations used; it was therefore judged suitable as a pH indicator. Intracellular pH was then measured by distribution of morpholine (6.985 ± 0.08), nicotine (6.915 ± 0.03) and the weak acid 5,5′-dimethyl-2,4-oxazolidinedione (7.10 ± 0.05) and with pH-sensitive microelectrodes (5.9, the equilibrium value). It was shown that the four significantly different values could not be reconciled in terms of experimental error, heterogeneity of intracellular pH, liquid junction potential differences, or binding of indicator molecules inside the fibre. They could, however, be reconciled if the fibre water had different structure and solvent properties from the extracellular water and ions were distributed across the membrane as between two liquid phases containing different solvents. Then the H⁺ would be in equilibrium, as shown by the microelectrode measurement, but intracellular pH would be indeterminable and probably greater than 6.     

The appearance of exo-α-mannosidase in cultures of Penicillium charlessi

Summary Growth of the fungus Penicillium charlesii G. Smith on glucose, minimal salts medium results in the appearance of -d-mannopyranosidase activity capable of hydrolyzing p-nitrophenyl--d-mannopyranoside. No activity is found until depletion of the carbon source, after which the enzyme activity rapidly increases in the mycelium. Prolonged incubation of the culture results in the appearance of small amounts of -mannosidase activity in the growth medium. The initial release of a mannose-containing polysaccharide into the medium precedes the appearance of -mannosidase by several days.     

Dimerization of CtIP may stabilize in vivo interactions with the Retinoblastoma-pocket domain

CtIP is a tumor suppressor that interacts with Retinoblastoma protein (Rb) to regulate the G1/S-phase transition of the cell cycle. Despite its large size (897 residues) CtIP has few known structured regions. Rather it contains several linear motifs that interact with known binding partners, including an LXCXE motif that binds the pocket domain of Rb-family proteins. This LXCXE motif lies at the C-terminus of the only known structured domain, an N-terminal coiled-coil dimerization domain (DD; residues 45-160). Yeast two-hybrid (Y2H) and GST-pulldown analyses showed that CtIP requires the LXCXE motif to bind the Rb-pocket. Although isothermal titration calorimetry data indicates that the LXCXE motif is the sole determinant of binding affinity for the Rb-pocket domain (K(A) approximately 10(6)M(-1)), Y2H data indicates that the DD is required to stabilize the interaction in vivo. Thus dimerization may increase the apparent stability of the proteins and/or the lifetime of the complexes.     

New Zealanders working non-standard hours also have greater exposure to other workplace hazards

Exposure to workplace hazards, such as dust, solvents, and fumes, has the potential to adversely affect the health of people. However, the effects of workplace hazards on health may differ when exposure occurs at different times in the circadian cycle, and among people who work longer hours or who do not obtain adequate sleep. The aim of the present study was to document exposures to workplace hazards across a national sample of New Zealanders, comparing people who work a standard 08:00 ?17:00 h Monday-to-Friday working week (Std hours) and those who do not (N-Std hours). New Zealanders (n = 10 000) aged 20–64 yrs were randomly selected from the Electoral Roll to take part in a nationwide survey of workplace exposures. Telephone interviews were conducted between 2004 and 2006, using a six-part questionnaire addressing demographics, detailed information on the current or most recent job (including exposures to a range of workplace hazards), sleep, sleepiness, and health status. N-Std hours were categorised on the basis of: being required to start work prior to 07:00 h or finish work after 21:00 h and/or; having a regular on-call commitment (at least once per week) and/or; working rotating shifts and/or; working night shift(s) in the last month. The response rate was 37% (n = 3003), with 22.2% of participants (n = 656) categorised as working N-Std hours. Industry sectors with the highest numbers of participants working N-Std hours were manufacturing, health and community services, and agriculture, fishing, and forestry. Response rate was 37% (n = 3003) with 22.2% (n = 656) categorised as working N-Std hours. Participants working N-Std hours were more likely to be exposed to all identified hazards, including multiple hazards (OR = 2.45, 95% CI = 2.01–3.0) compared to those working Std hours. Participants working N-Std hours were also more likely to report ‘never/rarely’ getting enough sleep (OR = 1.38, 95% CI = 1.15–1.65), ‘never/rarely’ waking refreshed (OR = 1.23, 95% CI = 1.04–1.47), and excessive sleepiness (OR = 1.77, 95% CI = 1.29–2.42). New Zealanders working N-Std hours are more likely to be exposed to hazards in the workplace, to be exposed to multiple hazards, and to report inadequate sleep and excessive sleepiness than their colleagues working a standard 08:00?17:00 h Monday-to-Friday working week. More research is needed on the effects of exposure to hazardous substances outside the usual waking day, on the effects of exposure to multiple hazards, and on the combination of hazard exposure and sleep restriction as a result of shift work.