Dynamic structure of lipid-bound synaptobrevin suggests a nucleation-propagation mechanism for trans-SNARE complex formation

The synaptic vesicle protein synaptobrevin engages with syntaxin and SNAP-25 to form the SNARE complex, which drives membrane fusion in neuronal exocytosis. In the SNARE complex, the SNARE motif of synaptobrevin forms a 55-residue helix, but it has been assumed to be mostly unstructured in its prefusion form. NMR data for full-length synaptobrevin in dodecylphosphocholine micelles reveals two transient helical segments flanked by natively disordered regions and a third more stable helix. Transient helix I comprises the most N-terminal part of the SNARE motif, transient helix II extends the SNARE motif into the juxtamembrane region, and the more stable helix III is the transmembrane domain. These helices may have important consequences for SNARE complex folding and fusion: helix I likely forms a nucleation site, the C-terminal disordered SNARE motif may act as a folding arrest signal, and helix II likely couples SNARE complex folding and fusion.     

The relationship between alcohol consumption and cortisol secretion in an aging cohort

CONTEXT: Evidence for an association between alcohol consumption and activity of the hypothalamic-pituitary-adrenal (HPA) axis is inconclusive. OBJECTIVE: Our objective was to assess the relationship between indices of alcohol consumption and salivary cortisol concentration. DESIGN: This was a cross-sectional study of alcohol consumption and cortisol secretion from phase 7 (2002-2004) of the Whitehall II study. SETTING: An occupational cohort originally recruited in 1985-1987 was included in the study. PARTICIPANTS: A total of 2693 men and 977 women had information on cortisol levels and alcohol consumption. OUTCOME MEASURES: Saliva samples were taken on waking, waking+0.5, 2.5, 8, and 12 h, and bedtime for the assessment of cortisol. RESULTS: In men there was a positive association between cortisol and units of alcohol intake per week (3% increase in cortisol per unit of alcohol consumed; P=0.010). The slope of cortisol decline over the day in heavy drinkers was reduced (heavy drinkers beta=-0.155, moderate drinkers beta=-0.151), indicating reduced control of the HPA axis in heavy drinkers. In women the cortisol awakening response was greater in heavy drinkers 14.15 nmol/liter (9.12-19.17) compared with moderate drinkers 8.69 nmol/liter (7.72-9.67) (P=0.037). CONCLUSIONS: This study suggests that alcohol consumption is associated with activation of the HPA axis. These results are not due to alcohol consumption on the day, suggesting chronic changes of the HPA axis in heavy drinking groups.     

Typing of Haemophilus influenzae by Coagglutination and Conventional Slide Agglutination

Coagglutination was compared with conventional slide agglutination for the typing of 297 clinical isolates of Haemophilus sp. A 100% correlation was found with the H. influenzae type b isolates. Coagglutination showed no false-positive reactions with the nontypable strains of H. influenzae and H. parainfluenzae isolates; however, conventional slide agglutination exhibited many false-positive and non-interpretable reactions.     

Vaccination of newborn mice induces a strong protective immune response against respiratory and genital challenges with Chlamydia trachomatis

Chlamydia trachomatis infections can occur early in life and may result in long-term sequelae. To assess the feasibility of implementing a vaccine in newborns, groups of 2-day-old BALB/c mice were immunized intranasally (i.n.) with 1x10(4) inclusion forming units (IFU) of C. trachomatis mouse pneumonitis (MoPn). As a control, newborn mice were sham-immunized i.n. with minimal essential medium. In the vaccinated animals, strong Chlamydia-specific humoral and cell-mediated immune responses were observed. Six weeks after immunization, mice were challenged with MoPn i.n. or intravaginally (i.vag.). For the i.n. challenge, mice were inoculated with 10(4) or 10(5)IFU of MoPn per mouse, and in the case of the i.vag. challenge, each animal received 10(6)IFU. By day 10 post-infection (p.i.), the vaccinated mice challenged i.n. with 10(4)IFU, had gained an average of 6.7+/-1% of their body weight. In contrast, the sham-immunized mice had lost 14.9+/-1% of their weight (P<0.05). The mean number of IFU/lungs in the vaccinated animals was 800+/-300, while for the sham-immunized mice was 211+/-49x10(6) (P<0.05). Significant differences between the Chlamydia-vaccinated and the sham-immunized mice were also found in the groups challenged with 10(5)IFU. In the mice challenged i.vag., a significant decrease in the number of mice with positive cultures, and the intensity and duration of vaginal shedding was noted in the vaccinated mice compared to the sham-immunized mice (P<0.05). In conclusion, these results indicate that vaccination of neonatal mice can result in a protective response against a subsequent pulmonary or genital challenge with Chlamydia.     

The relationship between smoking status and cortisol secretion

CONTEXT: Evidence for an association of smoking status with cortisol secretion is mixed. OBJECTIVE: The objective of the study was to assess the relationship between smoking status and salivary cortisol. DESIGN: This was a cross-sectional study of smoking status and cortisol secretion from phase 7 (2002-2004) of the Whitehall II study. SETTING: An occupational cohort was originally recruited in 1985-1987. PARTICIPANTS: The study population consisted of 3103 men (1514 never-smokers, 1278 ex-smokers, and 311 smokers) and 1128 women (674 never-smokers, 347 ex-smokers, and 107 smokers). Information was collected on smoking status, average number of cigarettes smoked, and additional covariates. OUTCOME MEASURES: Saliva samples were taken on waking; waking + 0.5, 2.5, 8, and 12 h; and bedtime for the assessment of cortisol. RESULTS: Smoking status was significantly associated with increased salivary cortisol release throughout the day (P < 0.001) adjusted for covariates; this was apparent for the cortisol awakening response (P < 0.001) when examined separately. Compared with never-smokers, smokers had higher release of total cortisol (P = 0.002), whereas no difference was observed between never-smokers and ex-smokers (P = 0.594): mean release per hour (nanomoles per liter), never-smokers, 4.13 [confidence interval (CI) 4.02-4.24]; ex-smokers, 4.21 (CI 4.08-4.35); smokers, 4.63 (CI 4.35-4.93). There was no significant relationship between number of cigarettes smoked and total cortisol release. However, a difference was observed for the cortisol awakening response: mean release by tertiles of cigarettes smoked (nanomoles per liter): high, 13.49 (CI 10.74-16.23); medium, 9.58 (CI 7.40-11.76); low, 8.49 (CI 5.99-10.99), P = 0.029. CONCLUSION: Salivary cortisol is increased in current smokers, compared with nonsmokers; no differences were observed between ex-smokers and never-smokers, suggesting that smoking has a short-term effect on the neuroendocrine system.     

Psychological coping styles and cortisol over the day in healthy older adults

Patterns of psychological coping are associated with a variety of health outcomes but the underlying pathways are not yet established. The purpose of this study was to assess the relationship between salivary cortisol output over the course of a day and coping style. Data were available from 350 men and 192 women with an average age of 60.9 years. Participants were drawn from the Whitehall II cohort, and had no history of cardiovascular disease. Individuals who were taking medication that might affect cortisol levels were also excluded. Saliva samples were provided on waking, then 0.5, 2.5, 8 and 12 h after waking, and just before the participant went to sleep. Coping style was measured with a standard instrument, the COPE, and data were factor analysed to generate three factors: seeking social support, problem engagement and problem avoidance. The relationships between these factors and the cortisol awakening response (CAR), the slope of cortisol change over the day and total cortisol output over the day (excluding the waking period) were assessed using multiple linear regression. Cortisol output over the day was inversely associated with coping with stress by seeking social support (p=0.034) and by problem engagement (p=0.003), independently of age, gender, body mass index, smoking, depression, self-rated health, time of waking and income. Individuals who coped by problem engagement and seeking support had lower cortisol levels. Additionally, gender, BMI, smoking, self-rated health and time of waking were independently related to cortisol output over the day. There were no significant associations between coping and the CAR or cortisol slope over the day. The results indicate that adaptive coping styles are related to low levels of cortisol over the day, suggesting that neuroendocrine pathways may partly mediate relationships between psychological coping and health.