Structural basis for myopathic defects engendered by alterations in the myosin rod

While mutations in the myosin subfragment 1 motor domain can directly disrupt the generation and transmission of force along myofibrils and lead to myopathy, the mechanism whereby mutations in the myosin rod influences mechanical function is less clear. Here, we used a combination of various imaging techniques and molecular dynamics simulations to test the hypothesis that perturbations in the myosin rod can disturb normal sarcomeric uniformity and, like motor domain lesions, would influence force production and propagation. We show that disrupting the rod can alter its nanomechanical properties and, in vivo, can drive asymmetric myofilament and sarcomere formation. Our imaging results indicate that myosin rod mutations likely disturb production and/or propagation of contractile force. This provides a unifying theory where common pathological cascades accompany both myosin motor and specific rod domain mutations. Finally, we suggest that sarcomeric inhomogeneity, caused by asymmetric thick filaments, could be a useful index of myopathic dysfunction.     

Molecular evolution: Please release me, genetic code

The genetic code is no longer universal, even in non-mitochondrial genomes. Recent studies have implicated the eukaryotic release factor eRF1 in mediating coding changes that are not as inconceivable as once thought. Specific residues in eRF1 proteins can be correlated with specific code changes in a wide variety of taxa.     

The Prevalence and Correlates of Lifetime Psychiatric Disorders and Trauma Exposures in Urban and Rural Settings: Results from the National Comorbidity Survey Replication (NCS-R)

Introduction

Distinctions between rural and urban environments produce different frequencies of traumatic exposures and psychiatric disorders. We examine the prevalence of psychiatric disorders and frequency of trauma exposures by position on the rural-urban continuum.

Methods

The National Comorbidity Survey Replication (NCS-R) was used to evaluate psychiatric disorders among a nationally-representative sample of the U.S. population. Rurality was designated using the Department of Agriculture''s 2003 rural-urban continuum codes (RUCC), which differentiate counties into levels of rurality by population density and adjacency to metropolitan areas. Lifetime psychiatric disorders included post-traumatic stress disorder (PTSD), anxiety disorders, major depressive disorder, mood disorders, impulse-control disorders, and substance abuse. Trauma exposures were classified as war-related, accident-related, disaster-related, interpersonal or other. Weighted logistic regression models examined the odds of psychiatric disorders and trauma exposures by position on the rural-urban continuum, adjusted for relevant covariates.

Results

75% of participants were metropolitan, 12.2% were suburban, and 12.8% were from rural counties. The most common disorder reported was any anxiety disorder (38.5%). Drug abuse was more common among metropolitan (8.7%, p = 0.018), compared to nonmetropolitan (5.1% suburban, 6.1% rural) participants. A one-category increase in rurality was associated with decreased odds for war-related trauma (aOR = 0.86, 95%CI 0.78–0.95). Rurality was not associated with risk for any other lifetime psychiatric disorders or trauma exposure.

Discussion/Conclusions

Contrary to the expectation of some rural primary care providers, the frequencies of most psychiatric disorders and trauma exposures are similar across the rural-urban continuum, reinforcing calls to improve mental healthcare access in resource-poor rural communities.     

SPECIES COMPOSITION AND SEASONAL PERIODICITY OF MACROALGAL SPECIES IN CORPUS CHRISTI BAY, TEXAS

Benthic algal sampling from selected sites along Corpus Christi Bay and from one site at the Port Aransas, Texas south jetty was completed between April 1999 and February, 2000. Species composition, seasonal periodicity, and fluctuations in temperature and salinity were determined. This is the first comprehensive study of benthic macroalgae conducted in Corpus Christi Bay, which is shallow, turbid, and lacks natural hard substrate. Man-made jetties are necessary for suitable floral attachment. Macroalgae are affected by changes in salinity as freshwater inflows are followed by periods of drought, which increase salinity. These effects are most notable where freshwater enters at the south end near Oso Bay and at the north end at Nueces Bay. Previous Texas algal collections described species of Enteromorpha, Ulva, Gelidium , and Gracilaria as the most dominant plants of the area. This supports the current study with the additions of Hypnea musciformis and Centroceras clavulatum. Dominant plants at the Port Aransas jetty include Ulva fasciata, Padina gymnospora , and Hypnea musciformis. The Rhodophyta including Gracilaria, Gelidium , and Centroceras clavulatum dominate the bay and do so throughout the year. Chlorophytes, although few in species richness, are important to benthic coverage and biomass. Phaeophyta are found predominantly at the Port Aransas jetty with Sargassum , Dictyota dichotoma , and Petalonia fascia being most abundant. A transition occurs in species composition as the water temperature changes seasonally. Hincksia, Ectocarpus , and Petalonia fascia are found only during the cooler months.     

Influenza H5N1 virus infection of polarized human alveolar epithelial cells and lung microvascular endothelial cells

Background

Highly pathogenic avian influenza (HPAI) H5N1 virus is entrenched in poultry in Asia and Africa and continues to infect humans zoonotically causing acute respiratory disease syndrome and death. There is evidence that the virus may sometimes spread beyond respiratory tract to cause disseminated infection. The primary target cell for HPAI H5N1 virus in human lung is the alveolar epithelial cell. Alveolar epithelium and its adjacent lung microvascular endothelium form host barriers to the initiation of infection and dissemination of influenza H5N1 infection in humans. These are polarized cells and the polarity of influenza virus entry and egress as well as the secretion of cytokines and chemokines from the virus infected cells are likely to be central to the pathogenesis of human H5N1 disease.

Aim

To study influenza A (H5N1) virus replication and host innate immune responses in polarized primary human alveolar epithelial cells and lung microvascular endothelial cells and its relevance to the pathogenesis of human H5N1 disease.

Methods

We use an in vitro model of polarized primary human alveolar epithelial cells and lung microvascular endothelial cells grown in transwell culture inserts to compare infection with influenza A subtype H1N1 and H5N1 viruses via the apical or basolateral surfaces.

Results

We demonstrate that both influenza H1N1 and H5N1 viruses efficiently infect alveolar epithelial cells from both apical and basolateral surface of the epithelium but release of newly formed virus is mainly from the apical side of the epithelium. In contrast, influenza H5N1 virus, but not H1N1 virus, efficiently infected polarized microvascular endothelial cells from both apical and basolateral aspects. This provides a mechanistic explanation for how H5N1 virus may infect the lung from systemic circulation. Epidemiological evidence has implicated ingestion of virus-contaminated foods as the source of infection in some instances and our data suggests that viremia, secondary to, for example, gastro-intestinal infection, can potentially lead to infection of the lung. HPAI H5N1 virus was a more potent inducer of cytokines (e.g. IP-10, RANTES, IL-6) in comparison to H1N1 virus in alveolar epithelial cells, and these virus-induced chemokines were secreted onto both the apical and basolateral aspects of the polarized alveolar epithelium.

Conclusion

The predilection of viruses for different routes of entry and egress from the infected cell is important in understanding the pathogenesis of influenza H5N1 infection and may help unravel the pathogenesis of human H5N1 disease.     

Should symptomatic menopausal women be offered hormone therapy?

Many physicians remain uncertain about prescribing hormone therapy for symptomatic women at the onset of menopause. The American Society for Reproductive Medicine (ASRM) convened a multidisciplinary group of healthcare providers to discuss the efficacy and risks of hormone therapy for symptomatic women, and to determine whether it would be appropriate to treat women at the onset of menopause who were complaining of menopausal symptoms. MAJOR FINDINGS: Numerous controlled clinical trials consistently demonstrate that hormone therapy, administered via oral, transdermal, or vaginal routes, is the most effective treatment for vasomotor symptoms. Topical vaginal formulations of hormone therapy should be preferred when prescribing solely for the treatment of symptoms of vulvar and vaginal atrophy. Data from the Women's Health Initiative indicate that the overall attributable risk of invasive breast cancer in women receiving estrogen plus progestin was 8 more cases per 10,000 women-years. No increased risk for invasive breast cancer was detected for women who never used hormone therapy in the past or for those receiving estrogen only. Hormone therapy is not effective for the treatment of cardiovascular disease and that the risk of cardiovascular disease with hormone therapy is principally in older women who are considerably postmenopause. CONCLUSIONS: Healthy symptomatic women should be offered the option of hormone therapy for menopausal symptoms. Symptom relief with hormone therapy for many younger women (at the onset of menopause) with menopausal symptoms outweighs the risks and may provide an overall improvement in quality of life. Hormone therapy should be individualized for symptomatic women. This involves tailoring the regimen and dose to individual needs.     

DNA polymerase-primase from embryos of Drosophila melanogaster. The DNA polymerase subunit

The DNA polymerase-primase from Drosophila melanogaster has been separated into its constituent polymerase and primase subunits by sedimentation in glycerol gradients containing 50% ethylene glycol. Both activities have been obtained in good yield. The properties of the 182-kDa polymerase subunit are similar to those of the intact four-subunit enzyme. However, there are three significant differences. (i) The polymerase activity of the 182-kDa subunit shows an increased thermolability; (ii) the pause sites during replication of singly primed, single-stranded circular DNA by the 182-kDa subunit are altered; and (iii) unlike the intact enzyme, the 182-kDa subunit is highly processive in the presence of the single-stranded DNA-binding protein of Escherichia coli.     

Structural basis for the regulation of muscle contraction by troponin and tropomyosin

The molecular switching mechanism governing skeletal and cardiac muscle contraction couples the binding of Ca²⁺ on troponin to the movement of tropomyosin on actin filaments. Despite years of investigation, this mechanism remains unclear because it has not yet been possible to directly assess the structural influence of troponin on tropomyosin that causes actin filaments, and hence myosin-crossbridge cycling and contraction, to switch on and off. A C-terminal domain of troponin I is thought to be intimately involved in inducing tropomyosin movement to an inhibitory position that blocks myosin-crossbridge interaction. Release of this regulatory, latching domain from actin after Ca²⁺ binding to TnC (the Ca²⁺ sensor of troponin that relieves inhibition) presumably allows tropomyosin movement away from the inhibitory position on actin, thus initiating contraction. However, the structural interactions of the regulatory domain of TnI (the “inhibitory” subunit of troponin) with tropomyosin and actin that cause tropomyosin movement are unknown, and thus, the regulatory process is not well defined. Here, thin filaments were labeled with an engineered construct representing C-terminal TnI, and then, 3D electron microscopy was used to resolve where troponin is anchored on actin-tropomyosin. Electron microscopy reconstruction showed how TnI binding to both actin and tropomyosin at low Ca²⁺ competes with tropomyosin for a common site on actin and drives tropomyosin movement to a constrained, relaxing position to inhibit myosin-crossbridge association. Thus, the observations reported reveal the structural mechanism responsible for troponin-tropomyosin-mediated steric interference of actin-myosin interaction that regulates muscle contraction.