'Like building a plane and flying it all in one go': an interview study of infection prevention and control in Australian general practice during the first 2 years of the SARS-CoV-2 pandemic.
General practitioners (GPs) and their staff have been at the frontline of the SARS-CoV-2 pandemic in Australia. However, their experiences of responding to and managing the risks of viral transmission within their facilities are poorly described. The aim of this study was to describe the experiences, and infection prevention and control (IPC) strategies adopted by general practices, including enablers of and challenges to implementation, to contribute to our understanding of the pandemic response in this critical sector. Semistructured interviews were conducted in person, by telephone or online video conferencing software, between November 2020 and August 2021. Twenty general practice personnel working in New South Wales, Australia, including nine GPs, one general practice registrar, four registered nurses, one nurse practitioner, two practice managers and two receptionists. Participants described implementing wide-ranging repertoires of IPC strategies-including telehealth, screening of patients and staff, altered clinic layouts and portable outdoor shelters, in addition to appropriate use of personal protective equipment (PPE)-to manage the demands of the SARS-CoV-2 pandemic. Strategies were proactive, influenced by the varied contexts of different practices and the needs and preferences of individual GPs as well as responsive to local, state and national requirements, which changed frequently as the pandemic evolved. Using the 'hierarchy of controls' as a framework for analysis, we found that the different strategies adopted in general practice often functioned in concert with one another. Most strategies, particularly administrative and PPE controls, were subjected to human variability and so were less reliable from a human factors perspective. However, our findings highlight the creativity, resilience and resourcefulness of general practice staff in developing, implementing and adapting their IPC strategies amidst constantly changing pandemic conditions.


#Science: The Potential and the Challenges of Utilizing Social Media and Other Electronic Communication Platforms in Health Care.
Electronic communication is becoming increasingly popular worldwide, as evidenced by its widespread and rapidly growing use. In medicine, however, it remains a novel approach to reach out to patients. Yet, they have the potential for further improving current health care. Electronic platforms could support therapy adherence and communication between physicians and patients. The power of social media as well as other electronic devices can improve adherence as evidenced by the development of the app bant. Additionally, systemic analysis of social media content by Screenome can identify health events not always captured by regular health care. By better identifying these healthcare events we can improve our current healthcare system as we will be able to better tailor to the patients' needs. All these techniques are a valuable component of modern health care and will help us into the future of increasingly digital health care.


Evidence for cascades of perturbation and adaptation in the metabolic genes of higher termite gut symbionts.
Termites and their gut microbes engage in fascinating dietary mutualisms. Less is known about how these complex symbioses have evolved after first emerging in an insect ancestor over 120 million years ago. Here we examined a bacterial gene, formate dehydrogenase (fdhF), that is key to the mutualism in 8 species of "higher" termite (members of the Termitidae, the youngest and most biomass-abundant and species-rich termite family). Patterns of fdhF diversity in the gut communities of higher termites contrasted strongly with patterns in less-derived (more-primitive) insect relatives (wood-feeding "lower" termites and roaches). We observed phylogenetic evidence for (i) the sweeping loss of several clades of fdhF that may reflect extinctions of symbiotic protozoa and, importantly, bacteria dependent on them in the last common ancestor of all higher termites and (ii) a radiation of genes from the (possibly) single allele that survived. Sweeping gene loss also resulted in (iii) the elimination of an entire clade of genes encoding selenium (Se)-independent enzymes from higher termite gut communities, perhaps reflecting behavioral or morphological innovations in higher termites that relaxed preexisting environmental limitations of Se, a dietary trace element. Curiously, several higher termite gut communities may have subsequently reencountered Se limitation, reinventing genes for Se-independent proteins via convergent evolution. Lastly, the presence of a novel fdhF lineage within litter-feeding and subterranean higher (but not other) termites may indicate recent gene "invasion" events. These results imply that cascades of perturbation and adaptation by distinct evolutionary mechanisms have impacted the evolution of complex microbial communities in a highly successful lineage of insects. Since patterns of relatedness between termite hosts are broadly mirrored by the relatedness of their symbiotic gut microbiota, coevolution between hosts and gut symbionts is rightly considered an important force that has shaped dietary mutualism since its inception over 120 million years ago. Apart from that concerning lateral gene or symbiont transfer between termite gut communities (for which no evidence yet exists), there has been little discussion of alternative mechanisms impacting the evolution of mutualism. Here we provide strong gene-based evidence for past environmental perturbations creating significant upheavals that continue to reverberate throughout the gut communities of species comprising a single termite lineage. We suggest that symbiont extinction events, sweeping gene losses, evolutionary radiations, relaxation and reemergence of key nutritional pressures, convergent evolution of similar traits, and recent gene invasions have all shaped gene composition in the symbiotic gut microbial communities of higher termites, currently the most dominant and successful termite family on Earth.


Constitutive telomerase expression promotes mammary carcinomas in aging mice.
Telomerase is up-regulated in the vast majority of human cancers and serves to halt the progressive telomere shortening that ultimately blocks would-be cancer cells from achieving a full malignant phenotype. In contrast to humans, the laboratory mouse possesses long telomeres and, even in early generation telomerase-deficient mice, the level of telomere reserve is sufficient to avert telomere-based checkpoint responses and to permit full malignant progression. These features in the mouse provide an opportunity to determine whether enforced high-level telomerase activity can serve functions that extend beyond its ability to sustain telomere length and function. Here, we report the generation and characterization of transgenic mice that express the catalytic subunit of telomerase (mTERT) at high levels in a broad variety of tissues. Expression of mTERT conferred increased telomerase enzymatic activity in several tissues, including mammary gland, splenocytes, and cultured mouse embryonic fibroblasts. In mouse embryonic fibroblasts, mTERT overexpression extended telomere lengths but did not prevent culture-induced replicative arrest, thus reinforcing the view that this phenomenon is not related to occult telomere shortening. Robust telomerase activity, however, was associated with the spontaneous development of mammary intraepithelial neoplasia and invasive mammary carcinomas in a significant proportion of aged females. These data indicate that enforced mTERT expression can promote the development of spontaneous cancers even in the setting of ample telomere reserve.


How important are direct fitness benefits of sexual selection?
Females may choose mates based on the expression of secondary sexual characters that signal direct, material fitness benefits or indirect, genetic fitness benefits. Genetic benefits are acquired in the generation subsequent to that in which mate choice is performed, and the maintenance of genetic variation in viability has been considered a theoretical problem. Consequently, the magnitude of indirect benefits has traditionally been considered to be small. Direct fitness benefits can be maintained without consideration of mechanisms sustaining genetic variability, and they have thus been equated with the default benefits acquired by choosy females. There is, however, still debate as to whether or not males should honestly advertise direct benefits such as their willingness to invest in parental care. We use meta-analysis to estimate the magnitude of direct fitness benefits in terms of fertility, fecundity and two measures of paternal care (feeding rate in birds, hatching rate in male guarding ectotherms) based on an extensive literature survey. The mean coefficients of determination weighted by sample size were 6.3%, 2.3%, 1.3% and 23.6%, respectively. This compares to a mean weighted coefficient of determination of 1.5% for genetic viability benefits in studies of sexual selection. Thus, for several fitness components, direct benefits are only slightly more important than indirect ones arising from female choice. Hatching rate in male guarding ectotherms was by far the most important direct fitness component, explaining almost a quarter of the variance. Our analysis also shows that male sexual advertisements do not always reliably signal direct fitness benefits.


Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature.
The conditional use of actin during clathrin-mediated endocytosis in mammalian cells suggests that the cell controls whether and how actin is used. Using a combination of biochemical reconstitution and mammalian cell culture, we elucidate a mechanism by which the coincidence of PI(4,5)P2 and PI(3)P in a curved vesicle triggers actin polymerization. At clathrin-coated pits, PI(3)P is produced by the INPP4A hydrolysis of PI(3,4)P2, and this is necessary for actin-driven endocytosis. Both Cdc42⋅guanosine triphosphate and SNX9 activate N-WASP-WIP- and Arp2/3-mediated actin nucleation. Membrane curvature, PI(4,5)P2, and PI(3)P signals are needed for SNX9 assembly via its PX-BAR domain, whereas signaling through Cdc42 is activated by PI(4,5)P2 alone. INPP4A activity is stimulated by high membrane curvature and synergizes with SNX9 BAR domain binding in a process we call curvature cascade amplification. We show that the SNX9-driven actin comets that arise on human disease-associated oculocerebrorenal syndrome of Lowe (OCRL) deficiencies are reduced by inhibiting PI(3)P production, suggesting PI(3)P kinase inhibitors as a therapeutic strategy in Lowe syndrome.


Are suicide rates higher in the cancer population? An investigation using forensic autopsy data.
Previous population-based studies have identified increased suicide rates among cancer patients. Available post mortem data on the contribution of cancer to completed suicide are limited, however. This study examines forensic autopsy data in order to assess whether cancer is significantly more frequent in a suicide population compared with a control population. In total, 232 cases were included in both the suicide and the control groups. Based on a complete standard histological examination, cancer was significantly more often found in the suicide group than in the control one (8.6% vs. 3.9%, p=0.03). The multivariate analysis also showed that the presence of cancer increased the risk of suicide. Moreover, cancer was not known to the deceased in 70% of cases, while the most frequent mental disease found in cancer-related suicide cases was depression (75%). In the 20 cancer-related suicide cases analysed herein, it was difficult to ascertain whether malignancy was the only motive for committing suicide, as cancer could be considered to be either a major causative factor for suicide or an incidental finding. Therefore, further research is warranted with the help of the psychological autopsy method to investigate the link between suicide and cancer further.


The relationship between anthropometric index and primary dysmenorehea in female high school students.
Primary dysmenorrhea, painful menstruation without pelvic pathologic complications, is one of the most common problems in women's gynecology and is one of the main causes for women referring to the clinic. Overweight and obesity may play a role in the etiology of primary dysmenorrhea. As a result, this study was conducted to determine the association between primary dysmenorrhea and anthropometric indices in female high school Students. This descriptive, cross-sectional study was conducted on 200 high school female students with primary dysmenorrhea in 2017 in Sabzevar. Anthropometric indicators were considered by the students if they had entry criteria. Data analysis was done using SPSS software. The results of data analysis showed that there was a significant difference between the presence of dysmenorrhea and anthropometric indices at height P = 0.05, waist circumference P = 0.03, waist circumference with height P = 0.01, height to waist circumference P = 0.01, thigh circumference to height P = 0.04, height to thigh circumference P = 0.04, waist circumference to hip circumference P = 0.04, hip circumference to hip circumference P = 0.05, and No significant relationship was found between the other indicators. Adherence to a balanced diet and proper lifestyle can prevent dysmenorrhea in adolescent girls.


[Aging and health: a change in paradigm].
The aging of the population due to increasing life spans requires a reevaluation of the health care models that prevail in the countries of the Region of the Americas, given that the illnesses typical of old age make up an ever-increasing share of the morbidity burden. The countries in the Region are in an intermediate stage of demographic transition. They face the challenge of balancing priority care for the health problems characteristic of developing countries along with care for the chronic and debilitating diseases that afflict the elderly. An even greater challenge is gauging, in a timely manner, the health, social, and economic consequences of the demographic transition over the next millennium, to allow the countries to respond to these consequences through an in-depth restructuring of their health and social services. The extension of human life imposes a new vision of the health of the elderly that is oriented toward self-sufficiency, economic independence, and living life to its fullest. It will not be easy to integrate this new paradigm into the countries' health services, which are now primarily oriented toward curative care and are structured to fulfill the requirements of an epidemiological scenario that is undergoing a rapid transformation. The challenge is even greater because of the wide range of needs that the elderly have across countries with varying degrees of development. Nevertheless, sooner or later the governments of all the countries of the Region will be forced to adopt concrete measures to deal with the problems posed by the population's gradual aging.


Casemix for inpatient care of elderly people: rehabilitation and post-acute care. Casemix for the Elderly Inpatient Working Group.
defining contracts for the care of elderly people on the basis of the number of episodes is inappropriate as it fails to take account of the wide variation in their physical disability and rehabilitation needs. Resource use on a day-to-day basis can be estimated for patients using the Resource Utilization Groups version III (RUG-III) casemix system. For practical use, RUG-III assessments cannot be made daily and so assessments at different time intervals were evaluated in order to give an indication of resource use for an inpatient stay. This study describes how RUG-III assessments can be used to give an indication of resource use for an inpatient episode. RUG-III assessments were completed for all admissions to elderly care rehabilitation wards in two Health Districts over a 10 week period. There were 336 patients and 965 RUG-III assessments. The average time required to make RUG-III assessments fell from 10 to 4 min by the end of the study period. Fortnightly assessment intervals including a discharge assessment correlated well with the average of weekly assessments (R2 = 0.88-0.91, P < 0.0001). using the results from these assessments we propose a model for use of the RUG-III system in contracts for rehabilitation and post-acute care of elderly people which addresses the difficulty of combining clinical characteristics, rehabilitation, resource use and length of stay into a single useful meaningful casemix system.


Phase change material based tunable reflectarray for free-space optical inter/intra chip interconnects.
The concept of phase change material (PCM) based optical antennas and antenna arrays is proposed for dynamic beam shaping and steering utilized in free-space optical inter/intra chip interconnects. The essence of this concept lies in the fact that the behaviour of PCM based optical antennas will change due to the different optical properties of the amorphous and crystalline state of the PCM. By engineering optical antennas or antenna arrays, it is feasible to design dynamic optical links in a desired manner. In order to illustrate this concept, a PCM based tunable reflectarray is proposed for a scenario of a dynamic optical link between a source and two receivers. The designed reflectarray is able to switch the optical link between two receivers by switching the two states of the PCM. Two types of antennas are employed in the proposed tunable reflectarray to achieve full control of the wavefront of the reflected beam. Numerical studies show the expected binary beam steering at the optical communication wavelength of 1.55 μm. This study suggests a new research area of PCM based optical antennas and antenna arrays for dynamic optical switching and routing.


A survey of medical ethics education at U.S. and Canadian medical schools.
To assess the format, content, method, and placement of medical ethics education in medical schools; the faculty and curricular resources and institutional structure and support of medical ethics; and the perceptions of ethics education among deans of medical education and medical ethics course directors at U.S. and Canadian medical schools. Two questionnaires were mailed to 125 U.S. medical schools and 16 Canadian schools: one to be completed by the deans of medical education and one to be completed by the medical ethics course director. Descriptive statistics were used to compare responses. In all, 123 (87%) deans and 91 (64%) course directors responded, providing information about 91 schools (six Canadian). All responding institutions offered some formal instruction in medical ethics, and among these, 71 (78%) incorporated ethics into required preclinical courses. The primary pedagogic course structure was small-group discussion and the primary pedagogic method was case discussions. One-fifth of schools provided no funding for ethics teaching, and 47 (52%) did not fund curricular development in ethics. Institutions with a dedicated ethics faculty member were twice as likely to have a mandatory introductory ethics course (64% versus 32%, p <.05). The primary obstacles to ethics education were thought to be a lack of time in the curriculum, a lack of qualified teachers, and a lack of time in faculty schedules. Within a few decades the number of U.S. and Canadian medical schools requiring medical ethics has increased. Nevertheless, significant variation in the content, method, and timing of ethics education suggests consensus about curricular content and pedagogic methods remains lacking. Further progress in ethics education may depend on institutions' willingness to devote more curricular time and funding to medical ethics.


Cytosol- and clathrin-dependent stimulation of endocytosis in vitro by purified adaptors.
Using stage-specific assays for receptor-mediated endocytosis of transferrin (Tfn) into perforated A431 cells we show that purified adaptors stimulate coated pit assembly and ligand sequestration into deeply invaginated coated pits. Late events in endocytosis involving membrane fission and coated vesicle budding which lead to the internalization of Tfn are unaffected. AP2, plasma membrane adaptors, are active at physiological concentrations, whereas AP1, Golgi adaptors, are inactive. Adaptor-dependent stimulation of Tfn sequestration requires cytosolic clathrin, but is unaffected by clathrin purified from coated vesicles suggesting that soluble and assembled clathrin pools are functionally distinct. In addition to adaptors and cytosolic clathrin other, as yet unidentified, cytosolic factors are also required for efficient coated pit invagination. These results provide new insight into the mechanisms and regulation of coated pit assembly and invagination.


Outcome of horses with synovial structure involvement following solar foot penetrations in four UK veterinary hospitals: 95 cases.
The factors associated with outcome following solar foot penetration involving synovial structures treated using endoscopic lavage have not been described in the UK population. To provide descriptive data on horses with synovial contamination or sepsis following solar penetration in 4 UK equine referral hospitals and to identify specific factors associated with the outcome. Retrospective case series. Data were collected from 4 veterinary hospitals. Follow-up data were obtained via a telephone questionnaire. Two multivariable logistic regression models were generated. Model 1 included all horses with synovial contamination following foot penetration undergoing surgical treatment, with the outcome variable being euthanasia during hospitalisation. Model 2 included all horses surviving anaesthesia, with the outcome variable being failure to return to pre-injury athletic function. Ninety-five horses were included. Overall, 56% of horses survived to discharge and 36% of horses returned to pre-injury athletic function. Model 1 included penetration of the central frog sulcus (odds ratio [OR] 10, 95% confidence interval [CI] 1.9-51.8), concurrent distal phalanx involvement (OR 32, 95% CI 2.6-101.9), increasing days to presentation (OR 1.2, 95% CI 1.0-1.3) and hospital. Model 2 included increasing days to presentation (OR 1.1, 95% CI 1.1-1.6), breed (OR 32, 95% CI 2.2-135.4), more than one surgery (OR 5.6, 95% CI 1.0-32.7) and hospital. Synovial involvement following solar foot penetration has a guarded prognosis for survival to discharge and a poor prognosis for return to pre-injury athletic function. Penetration of the central sulcus of the frog and distal phalanx involvement are associated with euthanasia during hospitalisation. Delayed referral and hospitalisation are associated with both euthanasia and failure to return to pre-injury athletic function. Breed and more than one surgery are associated with failure to return to pre-injury athletic function. These data may assist veterinary surgeons and owners to make evidence-based decisions when managing cases with synovial involvement following solar foot penetration.


Bone resorption inhibitor alendronate normalizes the reduced bone thickness of TRPV5(-/-) mice.
TRPV5 is a Ca(2+)-selective channel involved in transcellular Ca(2+) absorption expressed in kidney and in the ruffled border of osteoclasts. Studies in hypercalciuric TRPV5 knockout (TRPV5(-/-)) mice, which display significantly increased vitamin D levels, showed that TRPV5 ablation increases number and size of osteoclasts but impairs osteoclast-mediated bone resorption. The latter is not in line with the observed decreased bone thickness in TRPV5(-/-) mice. Bisphosphonates also inhibit osteoclast-mediated bone resorption. The aim of this study was to evaluate the effect of alendronate on the expression of the Ca(2+) transporters in bone, kidney, and duodenum and, importantly, the bone phenotype in TRPV5(-/-) mice. Wildtype (TRPV5(+/+)) and TRPV5(-/-) mice were treated during 10 wk with 2 mg/kg alendronate or vehicle weekly and housed in metabolic cages at the end of treatment. Urine and blood samples were taken for biochemical analysis, and duodenum, kidney, and femur were sampled. Expression of Ca(2+) transporters and osteoclast ruffled border transporters in bone and cultured osteoclasts was determined by QPCR analysis. Femurs were scanned using muCT, and resorption pit assays were performed in bone marrow cultures isolated from TRPV5(+/+) and TRPV5(-/-) mice. Alendronate treatment enhanced bone thickness in TRPV5(+/+) mice but also normalized the disturbed bone morphometry parameters in TRPV5(-/-) mice. Bone TRPV5 expression was specifically enhanced by alendronate, whereas the expression of Ca(2+) transporters in kidney and intestine was not altered. The expression of the osteoclast ruffled border membrane proteins chloride channel 7 (CLC-7) and the vacuolar H(+)-ATPase did not differ between both genotypes, but alendronate significantly enhanced the expression and PTH levels in TRPV5(-/-) mice. The expression of TRPV5, CLC-7, and H(+)-ATPase in osteoclast cultures was not affected by alendronate. The number of resorption pits was reduced in TRPV5(-/-) bone marrow cultures, but the response to vitamin D was similar to that in TRPV5(+/+) cultures. The alendronate-induced upregulation of TRPV5 in bone together with the decreased resorptive capacity of TRPV5(-/-) osteoclasts in vitro suggests that TRPV5 has an important role in osteoclast function. However, our data indicate that significant bone resorption still occurs in TRPV5(-/-) mice, because alendronate treatment normalized bone thickness in these mice. Thus, TRPV5(-/-) mice are able to rescue the resulting defect in osteoclast-mediated bone resorption, possibly mediated by the long-term hypervitaminosis D or other (non)hormonal compensatory mechanisms.


A novel tubulin polymerization inhibitor, MPT0B206, downregulates Bcr-Abl expression and induces apoptosis in imatinib-sensitive and imatinib-resistant CML cells.
Imatinib, a Bcr-Abl-specific inhibitor, is effective for treating chronic myeloid leukemia (CML), but drug resistance has emerged for this disease. In this study, we synthesized a novel tubulin polymerization inhibitor, MPT0B206 (N-[1-(4-methoxy-benzenesulfonyl)-2,3-dihydro-1H-indol-7-yl]-formamide), and demonstrated its apoptotic effect and mechanism in imatinib-sensitive K562 and imatinib-resistant K562R CML cells. Western blotting and immunofluorescence microscopy showed that MPT0B206 induced microtubule depolymerization in K562 and K562R cells. MPT0B206 inhibited the growth of these cells in a concentration- and time-dependent manner. It did not affect the viability of normal human umbilical vein endothelial cells. MPT0B206 induced G2/M cell cycle arrest and the appearance of the mitotic marker MPM-2 in K562 and K562R cells, which is associated with the upregulation of cyclin B1 and the dephosphorylation of Cdc2. Treatment of K562 and K562R cells with MPT0B206 induced apoptosis and reduced the protein levels of procaspase-9 and procaspase-3 and increased caspase-3 activity and PARP cleavage. MPT0B206 also reduced the levels of the antiapoptotic proteins Mcl-1 and Bcl-2 and increased the level of the apoptotic protein Bax. Additional experiments showed that MPT0B206 markedly downregulated Bcr-Abl mRNA expression and total and phosphorylated Bcr-Abl protein levels and inhibited the phosphorylation of its downstream proteins STAT5, MAPK, and AKT, and the protein level of c-Myc in K562 and K562R cells. Furthermore, MPT0B206 triggered viability reduction and apoptosis in CML cells carrying T315I-mutated Bcr-Abl. Together, these results suggest that MPT0B206 is a promising alternative for treating imatinib-resistant CML.


State health insurance market reforms and access to insurance for high-risk employees.
A specific focus for state regulations of the small group insurance market was to increase offers and stabilize premiums for firms with high-risk workers. We examine the effect of reforms implemented from 1993 through 1996 on the likelihood of employer sponsored insurance coverage. We find that packages of reforms that included both guaranteed issue of some products and some form of rate variance restriction had significant positive effects (4.5 percentage points) on ESI coverage for high-risk compared with low-risk workers within small firms and a small negative effect (-1.7 percentage points) on low-risk workers in small compared with large firms. The mechanism for these effects was an increase in take-up, rather than offer. Reform packages that included both guaranteed issue of all products and rate variance restrictions had similar effects overall, although they did not meet criteria for significance. These effects seemed to act through increased offer rather than take-up.


Widening educational disparities in premature death rates in twenty six states in the United States, 1993-2007.
Eliminating socioeconomic disparities in health is an overarching goal of the U.S. Healthy People decennial initiatives. We present recent trends in mortality by education among working-aged populations. Age-standardized death rates and their average annual percent change for all-cause and five major causes (cancer, heart disease, stroke, diabetes, and accidents) were calculated from 1993 through 2007 for individuals aged 25-64 years by educational attainment as a marker of socioeconomic status, using national vital registration data for 26 states with consistent educational information on the death certificates. Rate ratios and rate differences were used to assess disparities (≤12 versus ≥16 years of education) for 1993 through 2007. From 1993 through 2007, relative educational disparities in all-cause mortality continued to increase among working-aged men and women in the U.S., due to larger decreases of mortality rates among the most educated coupled with smaller decreases or even worsening trends in the less educated. For example, the rate ratios of all-cause mortality increased from 2.5 (95% confidence interval (CI), 2.4-2.6) in 1993 to 3.6 (95% CI, 3.5-3.7) in 2007 in men and from 1.9 (95% CI, 1.8-2.0) to 3.0 (95% CI, 2.9-3.1) in women. Generally, the rate differences (per 100,000 persons) of all-cause mortality increased from 415.5 (95% CI, 399.1-431.9) in 1993 to 472.7 (95% CI, 460.2-485.2) in 2007 in men and from 165.4 (95% CI, 154.5-176.2) to 256.2 (95% CI, 248.3-264.2) in women. Disparity patterns varied largely across the five specific causes considered in this study, with the largest increases of relative disparities for accidents, especially in women. Relative educational differentials in mortality continued to widen among men and women despite emphasis on reducing disparities in the U.S. Healthy People decennial initiatives.


Prenatal exposure to environmental insults and enhanced risk of developing Schizophrenia and Autism Spectrum Disorder: focus on biological pathways and epigenetic mechanisms.
When considering neurodevelopmental disorders (NDDs), Schizophrenia (SZ) and Autism Spectrum Disorder (ASD) are considered to be among the most severe in term of prevalence, morbidity and impact on the society. Similar features and overlapping symptoms have been observed at multiple levels, suggesting common pathophysiological bases. Indeed, recent genome-wide association studies (GWAS) and epidemiological data report shared vulnerability genes and environmental triggers across the two disorders. In this review, we will discuss the possible biological mechanisms, including glutamatergic and GABAergic neurotransmissions, inflammatory signals and oxidative stress related systems, which are targeted by adverse environmental exposures and that have been associated with the development of SZ and ASD. We will also discuss the emerging role of the gut microbiome as possible interplay between environment, immune system and brain development. Finally, we will describe the involvement of epigenetic mechanisms in the maintenance of long-lasting effects of adverse environments early in life. This will allow us to better understand the pathophysiology of these NDDs, and also to identify novel targets for future treatment strategies.


Negative Differential Conductance & Hot-Carrier Avalanching in Monolayer WS2 FETs.
The high field phenomena of inter-valley transfer and avalanching breakdown have long been exploited in devices based on conventional semiconductors. In this Article, we demonstrate the manifestation of these effects in atomically-thin WS2 field-effect transistors. The negative differential conductance exhibits all of the features familiar from discussions of this phenomenon in bulk semiconductors, including hysteresis in the transistor characteristics and increased noise that is indicative of travelling high-field domains. It is also found to be sensitive to thermal annealing, a result that we attribute to the influence of strain on the energy separation of the different valleys involved in hot-electron transfer. This idea is supported by the results of ensemble Monte Carlo simulations, which highlight the sensitivity of the negative differential conductance to the equilibrium populations of the different valleys. At high drain currents (>10 μA/μm) avalanching breakdown is also observed, and is attributed to trap-assisted inverse Auger scattering. This mechanism is not normally relevant in conventional semiconductors, but is possible in WS2 due to the narrow width of its energy bands. The various results presented here suggest that WS2 exhibits strong potential for use in hot-electron devices, including compact high-frequency sources and photonic detectors.


Interaction of subtype-selective antagonists with alpha 1-adrenergic receptor-mediated second messenger responses in rat brain.
The selective antagonists (+)-niguldipine and 5-methylurapidil (5-MU) were used to more clearly identify the alpha 1-adrenergic receptor subtypes involved in second messenger responses in slice and culture preparations of rat brain. The alpha 1-adrenergic receptor activating [3H]inositol phosphate (InsP) formation in neocortical and hippocampal slices appeared to have mixed characteristics. Although the low potency of (+)-niguldipine indicated involvement of the alpha 1B subtype, 5-MU had an alpha 1A-like potency at this subtype. (+)-Niguldipine did not inhibit the alpha 1 receptor-mediated potentiation of the cAMP response to either isoproterenol or adenosine in cortical slices, even at high concentrations. 5-MU inhibited both cAMP responses, although this inhibition appeared non-competitive. Thus, these receptors are clearly different from those mediating InsP formation. In primary glial cultures, (+)-niguldipine also had a low potency in blocking norepinephrine-stimulated [3H]InsP formation, consistent with involvement of the alpha 1B subtype. However, both 5-MU and WB 4101 had high potencies in blocking this response, suggesting involvement of the alpha 1A subtype. Inactivation of the alpha 1B subtype by pretreatment of cultures with chloroethylclonidine did not increase the potencies of any of these antagonists. The inhibition by 5-MU and WB 4101 was competitive in both control and chloroethylclonidine-pretreated cultures, whereas the inhibition by (+)-niguldipine was primarily noncompetitive. The use of these more selective antagonists shows that the current alpha 1A/alpha 1B subclassification scheme is inadequate to identify the receptors mediating these responses. None of the responses were blocked by (+)-niguldipine with the high potency expected at the alpha 1A subtype, although all InsP responses were blocked by 5-MU with a relatively high (alpha 1A-like) potency. In addition, very low affinity and noncompetitive effects of (+)-niguldipine were observed. These data raise the possibility of additional subtypes of alpha 1-adrenergic receptors or as yet unidentified functional interactions between known subtypes.


[Effect of biapenem (L-627) on fecal flora in gnotobiotic mice and children].
Biapenem (L-627), a novel injectable carbapenem antibiotic, was studied with regard to its effect on mice inoculated with four types of bacteria and on the intestinal flora of pediatric patients. L-627 was given i.m., 40 mg/kg once daily for 5 consecutive days, to mice inoculated enterically with four types of bacteria (Escherichia coli, Enterococcus faecalis, Bacteroides fragilis, and Bifidobacterium breve). Except for a mild decrease in E. coli, there were no major fluctuations in viable bacterial counts in the feces during the treatment. Five children with bacterial infections (3 boys and 2 girls; ages: 1 month to 7 years and 7 months; body weights 4.62-21.8 kg) were given L-627 at 6.0 to 11.7 mg/kg 3 times daily for 7 to 11 days. Among aerobes, although Enterobacteriaceae such as E. coli tended to decrease remarkably in all patients, there was no major change in Enterococcus. Consequently, total aerobe counts did not change significantly in any patient. Among anaerobes, Bifidobacterium, Bacteroides, and Eubacterium, which are the predominant organisms in infants, decreased remarkably in some patients. One of the patients showed a marked decrease in total anaerobe count associated with a change in fecal characteristics (diarrhea). Glucose nonfermenting Gram-negative bacilli or fungi did not become predominant organisms in any patient. Recovery from these changes in the intestinal flora was noted promptly after terminating L-627 treatment. L-627 was detected in the feces of 4 patients during treatment. The fecal concentration ranged from 0.24 to 2.22 micrograms/g. Clostridium difficile was not detected in any patient. Although C. difficile D-1 antigen was observed in 2 patients, it bore no relationship to fecal properties. The results indicated that L-627 had relatively few effects on the intestinal flora compared to other new beta-lactam antibiotics.


Structure-based interpretation of missense mutations in Y-family DNA polymerases and their implications for polymerase function and lesion bypass.
Our understanding of the molecular mechanisms of error-prone lesion bypass has changed dramatically in the past few years. The concept that the key participants in the mutagenic process were accessory proteins that somehow modified the ability of the cell's main replicase to facilitate bypass of normally blocking lesions has been replaced with one in which the replicase is displaced by a polymerase specialized in lesion bypass. The participants in this process remain the same, only their function has been reassigned. What was once known as the UmuC/DinB/Rev1/Rad30 superfamily of mutagenesis proteins, is now known as the Y-family of DNA polymerases. Quite remarkably, within the space of 3 years, the field has advanced from the initial discovery of intrinsic polymerase function, to the determination of the tertiary structures of several Y-family DNA polymerases.A key to determining the biochemical properties of each DNA polymerase is through structure-function studies that result in the site-specific substitution of particular amino acids at critical sites within each DNA polymerase. However, we should not forget the power of genetic selection that allows us to identify residues within each polymerase that are generated by "random mutagenesis" and which are important for both a gain or loss of function in vivo. In this review, we discuss the structural ramifications of several missense mutations previously identified in various Y-family DNA polymerase and speculate on how each amino acid substitution might modify the enzymatic activity of the respective polymerase or possibly perturb protein-protein interactions necessary for efficient translesion replication in vivo.


Visualization and quantification of intestinal transit and motor function by real-time tracking of 19F labeled capsules in humans.
A combined (19)F and (1)H MRI framework for the assessment of human intestinal transit and motor function is presented. This framework consists of silicone coated polychlorotrifluoroethylene capsules filled with perfluoro-[15]-crown-5-ether as (19)F marker, a flexible (19)F surface coil and a (19)F projection imaging sequence, allowing for real-time tracking of a single or multiple capsules. The capsules (length 11.5 mm, Ø 7.2 mm) contain 140 μL perfluoro-[15]-crown-5-ether and were tested for cytotoxicity and leakage prior to oral administration. A balanced SSFP projection sequence was implemented, yielding a temporal resolution of 133 ms. Optional multi-frequency excitation, allowing for interleaved tracking of differently labeled (19)F capsules, was incorporated. The passage of the (19)F capsules through intestinal sections was monitored in two healthy volunteers. Capsule coordinates were successfully coregistered with anatomical reference scans. Intestinal motility, residence times, lengths and forward velocities were determined. Simultaneous tracking of two capsules allowed for the assessment of peristaltic patterns with correction for respiratory motion. By providing the means for real-time multiple capsule tracking and high resolution anatomical imaging, the presented multinuclear imaging framework has the potential to provide important supplemental information for physiological and pharmaceutical research.


Antiatherogenic effect of captopril in the Watanabe heritable hyperlipidemic rabbit.
The effects of 9 months of orally administered captopril (25-50 mg/kg body wt/day) on aortic atherosclerosis was examined in normotensive Watanabe heritable hyperlipidemic rabbits. Captopril caused a significant decrease in aortic atherosclerosis. Total aortic surface involvement by lesions was reduced from 48 +/- 3.6% in control Watanabe rabbits to 30 +/- 3.9% with captopril treatment (p less than 0.01). Most of the decrease could be accounted for by a marked reduction in atherosclerosis of descending thoracic aortas from 49 +/- 5.2% to 15 +/- 3.9% in control and captopril-related groups, respectively (p less than 0.001). Significant decrease in cholesterol content of descending thoracic aorta was also observed in captopril-treated rabbits. Microscopic examination of the arterial lesions in captopril-treated animals suggested a relative decrease in cellularity and increase in extracellular matrix as compared with untreated animals. These studies indicate that captopril has a potent antiatherosclerotic action in the Watanabe heritable hyperlipidemic rabbit.


Emergence of large-scale cell morphology and movement from local actin filament growth dynamics.
Variations in cell migration and morphology are consequences of changes in underlying cytoskeletal organization and dynamics. We investigated how these large-scale cellular events emerge as direct consequences of small-scale cytoskeletal molecular activities. Because the properties of the actin cytoskeleton can be modulated by actin-remodeling proteins, we quantitatively examined how one such family of proteins, enabled/vasodilator-stimulated phosphoprotein (Ena/VASP), affects the migration and morphology of epithelial fish keratocytes. Keratocytes generally migrate persistently while exhibiting a characteristic smooth-edged "canoe" shape, but may also exhibit less regular morphologies and less persistent movement. When we observed that the smooth-edged canoe keratocyte morphology correlated with enrichment of Ena/VASP at the leading edge, we mislocalized and overexpressed Ena/VASP proteins and found that this led to changes in the morphology and movement persistence of cells within a population. Thus, local changes in actin filament dynamics due to Ena/VASP activity directly caused changes in cell morphology, which is coupled to the motile behavior of keratocytes. We also characterized the range of natural cell-to-cell variation within a population by using measurable morphological and behavioral features--cell shape, leading-edge shape, filamentous actin (F-actin) distribution, cell speed, and directional persistence--that we have found to correlate with each other to describe a spectrum of coordinated phenotypes based on Ena/VASP enrichment at the leading edge. This spectrum stretched from smooth-edged, canoe-shaped keratocytes--which had VASP highly enriched at their leading edges and migrated fast with straight trajectories--to more irregular, rounder cells migrating slower with less directional persistence and low levels of VASP at their leading edges. We developed a mathematical model that accounts for these coordinated cell-shape and behavior phenotypes as large-scale consequences of kinetic contributions of VASP to actin filament growth and protection from capping at the leading edge. This work shows that the local effects of actin-remodeling proteins on cytoskeletal dynamics and organization can manifest as global modifications of the shape and behavior of migrating cells and that mathematical modeling can elucidate these large-scale cell behaviors from knowledge of detailed multiscale protein interactions.


Why Virginity Pledges Succeed or Fail: The Moderating Effect of Religious Commitment Versus Religious Participation.
Over the past two decades, virginity pledges have proliferated in the US, despite mixed results regarding their effectiveness. Few studies have examined possible mechanisms that may shed light on why pledges work for some individuals but not others. Using a sample of emerging-adults aged 18-24 years old (n = 1,380),we examine the influence of religiosity on pledge signing and adherence, specifically whether the effectiveness of pledges is moderated by religiosity. Findings show that while religious participation is positively associated with signing a pledge, there is amoderating effect of religious commitment. That is, when religious commitment is high, adherence to the pledge is greater. However, for pledge signers with low religious commitment, there are unintended negative consequences with regard to increased participation in risky sexual behaviors, whether compared to other people who signed the pledge who are equally committed to their religion or to individuals who have never taken such a pledge. Implications for research and policy are discussed.


Aligning 3D time-of-flight MRA datasets for quantitative longitudinal studies: evaluation of rigid registration techniques.
3D Time-of-flight (TOF) magnetic resonance angiography is commonly used for vascular analyses. A quantification of longitudinal morphological changes usually requires the registration of TOF image sequences acquired at different time points. The aim of this study was to evaluate the precision of different 3D rigid registration setups such that an optimal quantification of morphological changes can be achieved. Eight different rigid registration techniques were implemented and evaluated in this study using the target registration error (TRE) calculated based on 554 landmarks defined in twenty TOF datasets. The registration techniques differed in integration of brain and vessel segmentation masks and usage of a multi-resolution framework. Furthermore, the benefit of a prior volume-of-interest definition for registration accuracy was evaluated. The results revealed that the highest registration accuracies can be achieved using a multi-resolution framework and a cerebrovascular segmentation as mask. Numerically, a mean TRE of 1.1mm was calculated. If applicable, a prior definition of a volume-of-interest allows a reduction of the TRE to only 0.6mm. TOF datasets should be registered using vessel segmentations as mask, multi-resolution framework and previous volume-of-interest definition if possible to obtain the highest registration precision. This is especially the case for longitudinal datasets that are separated by several months while the registration technique seems less important for datasets that are only separated by a few days.


cGMP inhibits the activity of single calcium channels in embryonic chick heart cells.
Effects of cGMP on the slow (L-type) Ca2+ channels of cultured chick embryonic cardiomyocytes were investigated by a cell-attached patch-clamp method. Superfusion of the single cells with 8-bromo-cGMP, a membrane-permeable derivative of cGMP, inhibited the single-channel activity. The cyclic nucleotide decreased, in a concentration-dependent manner, the ensemble averaged currents obtained from multichannel patches. 8-Bromo-cGMP (1 mM) completely abolished the currents (n = 8), whereas 0.1 mM only slightly decreased the currents (n = 4). The influence of cGMP on the characteristics of the single Ca2+ channels was examined using 0.3 mM 8-bromo-cGMP. Unit amplitude and slope conductance of the Ca2+ channel was not changed (25 pS in control versus 24 pS in the presence of cGMP). Analysis of single-channel kinetics showed that cGMP prolonged the slow time constant for the closed-time histogram (from 6.7 to 15.4 msec); the other time constants (for the open-time and closed-time histograms) were not affected. cGMP-induced inhibition of the Ca2+ channels may be mediated by cGMP-dependent protein kinase, because 8-bromo-cGMP is a potent activator of this protein kinase and does not stimulate cAMP hydrolysis. The present results suggest that cGMP opposes the effects of cAMP on the L-type Ca2+ channels in myocardial cells.


Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9.
CRISPR-Cas9-based genetic screens are a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target different sites in the genome with relative ease, but the on-target activity and off-target effects of individual sgRNAs can vary widely. Here, we use recently devised sgRNA design rules to create human and mouse genome-wide libraries, perform positive and negative selection screens and observe that the use of these rules produced improved results. Additionally, we profile the off-target activity of thousands of sgRNAs and develop a metric to predict off-target sites. We incorporate these findings from large-scale, empirical data to improve our computational design rules and create optimized sgRNA libraries that maximize on-target activity and minimize off-target effects to enable more effective and efficient genetic screens and genome engineering.


Adolescent preferences and reactions to language about body weight.
Over 30% of youth and adolescents have overweight or obesity, and health care providers are increasingly discussing weight-based health with these patients. Stigmatizing language in provider-patient communication about obesity is well documented and could be particularly detrimental to youth and adolescents. Although some research has examined preferences for weight-based terminology among adults, no studies have addressed these issues in youth populations. This study represents a preliminary and systematic investigation of weight-based language preferences among adolescents with overweight and obesity enrolled in a summer weight loss camp. Participants (N=50) indicated preferences for weight-based language and emotional responses to words that their family members used in reference to their body weight. Weight neutral terminology ('weight', 'body mass index') were most preferred, although some differences in word preferences emerged by the participants' gender. Boys preferred having their weight described as 'overweight' and 'heavy', while girls preferred the word 'curvy'. A large proportion of participants, particularly girls, reported experiencing sadness, shame, and embarrassment if parents used certain words to describe their body weight, which highlights the importance of considering the emotional impact of weight-based terminology. Providers may consider asking youth and adolescents for their preferences when discussing weight-based health.


Connection, Contagion, and COVID-19.
Anthropologists have long known how perceptions of contagion play out along the lines of xenophobia and racism. Months after the beginning of the global Cover-19 pandemic, predictions by anthropologists of xenophobic ideologies and actions have come to pass. In the United States people understand the global pandemic not as biology, but as the manifestation of political affiliation, difference, connection, and disconnection. COVID-19 is, according to public perception, dangerous because it maliciously mutates to attack. It is "a guy we don't know." Relationships between the mysteriousness of the virus and heightened visibility of longstanding inequality in the United States form new contexts for existing social tensions. These dynamics provide a backdrop against which the ongoing commitment to uprisings connected to the Black Lives Matter movement unfold. Here I draw on analysis of 50 semi-structured interviews we conducted from March to August of 2020 demonstrating how understandings of the biology of a virus are woven into perceptions of politics, inequality, and the fractures of a divided nation. To understand social and political responses to the global pandemic it is essential that we continue to investigate xenophobia, inequality, and racism alongside the biological impact of SARS-CoV-2.


Pleural complications of nasoenteric feeding tubes.
Pliable, small-bore, feeding tubes are inserted frequently in critically ill patients. The use of wire stylets to facilitate passage of these tubes has increased the chances of unrecognized tracheal intubations, particularly in obtunded patients. Five patients with depressed sensoria had inadvertent transbronchial insertions of feeding tubes into their pleural spaces over a 28-month period. The incidence of this complication among intensive care unit patients was 0.3%. Sequellae included pneumothoraces in four patients, pleural effusions in three patients, and pneumonia with a pulmonary abscess in one patient. The presence of cuffed endotracheal tubes did not prevent this complication in patients with depressed cough reflexes. Inexperience of feeding tube insertors may have contributed to pleural intubations in four of the patients. In patients with depressed sensoria, adequate confirmation of feeding tube location in the stomach or duodenum requires both chest and abdominal roentgenograms.


Melatonin promotes sleep-like state in zebrafish.
The sleep-promoting effect of the pineal hormone melatonin in humans is known for decades. However, the mechanisms of this phenomenon remain obscure, mainly due to lack of a simple, genetically tractable, animal model. We now report that melatonin promotes sleep-like state in a diurnal lower vertebrate, zebrafish (Danio rerio), and this effect is mediated through activation of specific melatonin membrane receptors. Furthermore, our data show that the sleep-like state in zebrafish has fundamental similarities with sleep in mammals, including characteristic postures, elevated arousal threshold to sensory stimulation and a compensatory rest rebound following rest deprivation, and can be induced by conventional hypnotics, diazepam and sodium pentobarbital. Collectively, these data indicate that melatonin is evolutionary conserved sleep-promoting agent in diurnal species and suggest that zebrafish provide an efficient animal model for studying the molecular mechanisms of sleep regulation and for screening new types of hypnotic medications.


Zinc requirement during meiosis I-meiosis II transition in mouse oocytes is independent of the MOS-MAPK pathway.
Zinc is essential for many biological processes, including proper functioning of gametes. We recently reported that zinc levels rise by over 50% during oocyte maturation and that attenuation of zinc availability during this period could be achieved using the membrane-permeable heavy metal chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN). This zinc insufficiency resulted in formation of large polar bodies, failure to establish metaphase II arrest, and impaired establishment of cortical polarity. As these phenotypes resemble those of MOS null oocytes, we examined the impact of zinc insufficiency on the MOS-MAPK pathway. Reduced levels of both MOS protein and phosphorylation of MAP2K1/2 are observed in zinc-insufficient oocytes; however, these differences appear only after completion of the first meiotic division. In addition, activation of the downstream effector of the MOS pathway, MAPK3/1, is not affected by zinc insufficiency, and reduced MOS levels are observed only with the presence of TPEN after the first polar body extrusion. These data are inconsistent with the hypothesis that reduced MOS mediates the observed phenotype. Finally, MOS overexpression does not rescue the phenotype of zinc-insufficient oocytes, confirming that the observed disruption of asymmetric division and spindle abnormalities cannot be attributed to impaired MOS signaling. Zinc-insufficient oocytes do not increase maturation promoting factor (MPF) activity following the first meiotic division, and increasing MPF activity through expression of nondegradable cyclin B1 partially rescues the ability of zinc-insufficient oocytes to enter metaphase II. Although we have shown that zinc has a novel role in the meiotic cell cycle, it is not mediated through the MOS-MAPK pathway.


Somatostatin receptor-targeted radionuclide therapy for progressive meningioma: benefit linked to 68Ga-DOTATATE/-TOC uptake.
The prognosis of patients with progressive meningioma after failure of surgery and radiotherapy is poor. We retrospectively evaluated the safety and efficacy of somatostatin-receptor (SSTR)-targeted radionuclide therapy (177Lu-DOTATATE [n = 16], 90Y-DOTATOC [n = 3], or both [n = 1]) in patients with progressive, treatment-refractory meningiomas (5 World Health Organization [WHO] grade I, 7 WHO grade II, 8 WHO grade III) and in part multifocal disease (17 of 20 patients). SSTR radionuclide treatment (median of 3 treatment cycles, median administered dose/cycle 7400 MBq) led to a disease stabilization in 10 of 20 patients for a median time of 17 months. Stratification according to WHO grade showed a median progression-free survival (PFS) of 32.2 months for grade I tumors, 7.2 for grade II, and 2.1 for grade III. PFS at 6 months was 100% for grade I, 57% for grade II, and 0% for grade III. Median overall survival was 17.2 months in WHO grade III patients and not reached for WHO I and II at a median follow-up of 20 months. In the analysis of single meningioma lesions, maximal and mean standardized uptake values in pretherapeutic 68Ga-DOTATOC/-TATE PET/CT were significantly higher in those lesions with radiographic stability after 6 months. In line with this, high expression of SSTR via immunohistochemistry was associated with PFS >6 months. SSTR-targeted radionuclide treatment has activity in a subset of patients with meningioma. Expression of SSTR via immunohistochemistry or radionuclide uptake might serve as a predictive biomarker for outcome to facilitate individualized treatment optimization in patients with uni- and multifocal meningiomas.


New binary direction aftereffect does not add up.
Neural adaptation and inhibition are pervasive characteristics of the primate brain and are probably understood better within the context of visual processing than with any other sensory modality. These processes are thought to underlie illusions in which one motion affects the perceived direction of another, such as the direction aftereffect (DAE) and direction repulsion. The DAE describes how, following prolonged viewing of motion in one direction, the direction of a subsequently viewed test pattern is misperceived. In the case of direction repulsion, the direction difference between two transparently moving surfaces is overestimated. Explanations of the DAE appeal to neural adaptation, whereas direction repulsion is accounted for through lateral inhibition. Here, we report on a new illusion, the binary DAE (bDAE), in which superimposed slow and fast dots moving in the same direction are perceived to move in different directions following adaptation to a mixed-speed stimulus. This new phenomenon is essentially a combination of the DAE and direction repulsion. Interestingly, the magnitude of the bDAE is greater than would be expected simply through a linear combination of the DAE and direction repulsion, suggesting that the mechanisms underlying these two phenomena interact in a nonlinear fashion.


Genome-wide ancestry of 17th-century enslaved Africans from the Caribbean.
Between 1500 and 1850, more than 12 million enslaved Africans were transported to the New World. The vast majority were shipped from West and West-Central Africa, but their precise origins are largely unknown. We used genome-wide ancient DNA analyses to investigate the genetic origins of three enslaved Africans whose remains were recovered on the Caribbean island of Saint Martin. We trace their origins to distinct subcontinental source populations within Africa, including Bantu-speaking groups from northern Cameroon and non-Bantu speakers living in present-day Nigeria and Ghana. To our knowledge, these findings provide the first direct evidence for the ethnic origins of enslaved Africans, at a time for which historical records are scarce, and demonstrate that genomic data provide another type of record that can shed new light on long-standing historical questions.


Spontaneous emission enhancement of a single molecule by a double-sphere nanoantenna across an interface.
We report on two orders of magnitude reduction in the fluorescence lifetime when a single molecule placed in a thin film is surrounded by two gold nanospheres across the film interface. By attaching one of the gold particles to the end of a glass fiber tip, we could control the modification of the molecular fluorescence at will. We find a good agreement between our experimental data and the outcome of numerical calculations.


Intercellular Transfer of Chromosomal Antimicrobial Resistance Genes between Acinetobacter baumannii Strains Mediated by Prophages.
The spread of antimicrobial resistance genes (ARGs) among Gram-negative pathogens, including Acinetobacter baumannii, is primarily mediated by transferable plasmids; however, ARGs are frequently integrated into its chromosome. How ARG gets horizontally incorporated into the chromosome of A. baumannii, and whether it functions as a cause for further spread of ARG, remains unknown. Here, we demonstrated intercellular prophage-mediated transfer of chromosomal ARGs without direct cell-cell interaction in A. baumannii We prepared ARG-harboring extracellular DNA (eDNA) components from the culture supernatant of a multidrug-resistant (MDR) A. baumannii NU-60 strain and exposed an antimicrobial-susceptible (AS) A. baumannii ATCC 17978 strain to the eDNA components. The antimicrobial-resistant (AR) A. baumannii ATCC 17978 derivatives appeared to acquire various ARGs, originating from dispersed loci of the MDR A. baumannii chromosome, along with their surrounding regions, by homologous recombination, with the ARGs including armA (aminoglycoside resistance), blaTEM-1 (β-lactam resistance), tet(B) (tetracycline resistance), and gyrA-81L (nalidixic acid resistance) genes. Notably, the eDNAs conferring antimicrobial resistance were enveloped in specific capsid proteins consisting of phage particles, thereby protecting the eDNAs from detergent and DNase treatments. The phages containing ARGs were likely released into the extracellular space from MDR A. baumannii, thereby transducing ARGs into AS A. baumannii, resulting in the acquisition of AR properties by the recipient. We concluded that the generalized transduction, in which phages were capable of carrying random pieces of A. baumannii genomic DNAs, enabled efficacious intercellular transfer of chromosomal ARGs between A. baumannii strains without direct cell-cell interaction.


Tendon neuroplastic training: changing the way we think about tendon rehabilitation: a narrative review.
Tendinopathy can be resistant to treatment and often recurs, implying that current treatment approaches are suboptimal. Rehabilitation programmes that have been successful in terms of pain reduction and return to sport outcomes usually include strength training. Muscle activation can induce analgesia, improving self-efficacy associated with reducing one's own pain. Furthermore, strength training is beneficial for tendon matrix structure, muscle properties and limb biomechanics. However, current tendon rehabilitation may not adequately address the corticospinal control of the muscle, which may result in altered control of muscle recruitment and the consequent tendon load, and this may contribute to recalcitrance or symptom recurrence. Outcomes of interest include the effect of strength training on tendon pain, corticospinal excitability and short interval cortical inhibition. The aims of this concept paper are to: (1) review what is known about changes to the primary motor cortex and motor control in tendinopathy, (2) identify the parameters shown to induce neuroplasticity in strength training and (3) align these principles with tendon rehabilitation loading protocols to introduce a combination approach termed as tendon neuroplastic training. Strength training is a powerful modulator of the central nervous system. In particular, corticospinal inputs are essential for motor unit recruitment and activation; however, specific strength training parameters are important for neuroplasticity. Strength training that is externally paced and akin to a skilled movement task has been shown to not only reduce tendon pain, but modulate excitatory and inhibitory control of the muscle and therefore, potentially tendon load. An improved understanding of the methods that maximise the opportunity for neuroplasticity may be an important progression in how we prescribe exercise-based rehabilitation in tendinopathy for pain modulation and potentially restoration of the corticospinal control of the muscle-tendon complex.


SENP1 enhances androgen receptor-dependent transcription through desumoylation of histone deacetylase 1.
SUMO (also called Sentrin) is a ubiquitin-like protein that plays an important role in regulating protein function and localization. It is known that several nuclear receptors are modified by SUMO; however, the effect of desumoylation in regulating nuclear receptor function has not been elucidated. Here we show that androgen receptor (AR)-mediated transcription is markedly enhanced by SENP1, a member of SUMO-specific protease family. SENP1's ability to enhance AR-dependent transcription is not mediated through desumoylation of AR, but rather through its ability to deconjugate histone deacetylase 1 (HDAC1), thereby reducing its deacetylase activity. HDAC1's repressive effect on AR-dependent transcription could be reversed by SENP1 and by deletion of its sumoylation sites. RNA interference depletion of endogenous HDAC1 also reduced SENP1's effect. Thus, SENP1 could regulate AR-dependent transcription through desumoylation of HDAC1. These studies provide insights on the potential role of desumoylation in the regulation of nuclear receptor activity.


Vitamin D3 supplementation and neurofilament light chain in multiple sclerosis.
Low circulating vitamin D levels are associated with an increased risk of active MRI lesions and relapses in several cohorts with relapsing remitting multiple sclerosis (RRMS). Randomized controlled supplementation trials are, however, negative on their primary endpoints, while secondary MRI endpoints suggest anti-inflammatory effects. Circulating levels of neurofilament light chain (NfL) are a biomarker of disease activity in RRMS. We explored whether 48-week high-dose vitamin D3 supplements were associated with lower circulating NfL levels. Of N = 40 Dutch interferon beta-treated participants with RRMS of the SOLAR trial, plasma samples at baseline and 48-week follow-up were available. Of these participants, N = 24 were supplemented with 14 000 IU/d vitamin D3 and N = 16 with placebo. Twenty-five hydroxyvitamin D3 (25(OH)D3 ) levels were measured with LC-MS/MS, and NfL levels were measured in duplicate with Simoa. Serum 25(OH)D3 levels at 48 weeks were increased in the vitamin D3 when compared to placebo group (median level 281 [IQR 205-330] vs 72 [39-88] nmol/L; P < .01). NfL levels at 48 weeks did not differ between the treatment groups (median level 25.4 [IQR 19.6-32.2] vs 25.3 [17.9-30.1] pg/mL; P = .74). Higher week 48 NfL level showed a trend toward association with a higher risk of combined unique active lesions on the week 48 MRI scan (OR 2.39 [95% CI 0.93-6.12] for each 10 pg/mL increase; P = .07). Supplementation of high-dose vitamin D3 for 48 weeks was not associated with lower NfL levels. This study does not support an effect of vitamin D3 on this biomarker of neuro-axonal injury.


An agent-based multilayer architecture for bioinformatics grids.
Due to the huge volume and complexity of biological data available today, a fundamental component of biomedical research is now in silico analysis. This includes modelling and simulation of biological systems and processes, as well as automated bioinformatics analysis of high-throughput data. The quest for bioinformatics resources (including databases, tools, and knowledge) becomes therefore of extreme importance. Bioinformatics itself is in rapid evolution and dedicated Grid cyberinfrastructures already offer easier access and sharing of resources. Furthermore, the concept of the Grid is progressively interleaving with those of Web Services, semantics, and software agents. Agent-based systems can play a key role in learning, planning, interaction, and coordination. Agents constitute also a natural paradigm to engineer simulations of complex systems like the molecular ones. We present here an agent-based, multilayer architecture for bioinformatics Grids. It is intended to support both the execution of complex in silico experiments and the simulation of biological systems. In the architecture a pivotal role is assigned to an "alive" semantic index of resources, which is also expected to facilitate users' awareness of the bioinformatics domain.


Symmetric TEM00 output from Q-switched quasi-concentric laser resonator with line-shaped end-pumping profile.
We report a 12.2 W pulsed TEM(00) output from Q-switched quasi-concentric laser resonator with line-shaped end-pumping profile, with the repetition rate of 30 kHz and optical-optical efficiency of 27.1%. The laser output mode is symmetrized in two directions in terms of beam quality, waist radius, and waist position.


Disclosure: What is the point and for whom?
Today mental health is embedded in a discourse of disclosure. Many people with lived experience of mental illness have decided to move out of the closet to talk about their personal experiences. To look at the context of disclosure and on the questions of why disclose and for whom. How much do we know and how does our knowledge correspond with today's discourse of disclosure in mental health? Narrative reviewing today's discourse of disclosure on the basis of both scientific and experience-based knowledge as well as from my personal experience. The scientific and experience-based knowledge of the benefits and costs of disclosure is limited, but points to mixed results and many risks on a personal level. At the same time, it leaves us with several unanswered questions. There is a need to always define the context of disclosure and to make professionals aware of different disclosure strategies and respond to the dilemmas. It is also crucial to emphasise that the planning and patterns of disclosure belong to the person with lived experience of mental illness and nobody else.


Sleep spindle and K-complex detection using tunable Q-factor wavelet transform and morphological component analysis.
A novel framework for joint detection of sleep spindles and K-complex events, two hallmarks of sleep stage S2, is proposed. Sleep electroencephalography (EEG) signals are split into oscillatory (spindles) and transient (K-complex) components. This decomposition is conveniently achieved by applying morphological component analysis (MCA) to a sparse representation of EEG segments obtained by the recently introduced discrete tunable Q-factor wavelet transform (TQWT). Tuning the Q-factor provides a convenient and elegant tool to naturally decompose the signal into an oscillatory and a transient component. The actual detection step relies on thresholding (i) the transient component to reveal K-complexes and (ii) the time-frequency representation of the oscillatory component to identify sleep spindles. Optimal thresholds are derived from ROC-like curves (sensitivity vs. FDR) on training sets and the performance of the method is assessed on test data sets. We assessed the performance of our method using full-night sleep EEG data we collected from 14 participants. In comparison to visual scoring (Expert 1), the proposed method detected spindles with a sensitivity of 83.18% and false discovery rate (FDR) of 39%, while K-complexes were detected with a sensitivity of 81.57% and an FDR of 29.54%. Similar performances were obtained when using a second expert as benchmark. In addition, when the TQWT and MCA steps were excluded from the pipeline the detection sensitivities dropped down to 70% for spindles and to 76.97% for K-complexes, while the FDR rose up to 43.62 and 49.09%, respectively. Finally, we also evaluated the performance of the proposed method on a set of publicly available sleep EEG recordings. Overall, the results we obtained suggest that the TQWT-MCA method may be a valuable alternative to existing spindle and K-complex detection methods. Paths for improvements and further validations with large-scale standard open-access benchmarking data sets are discussed.


The Vulnerability of Study Participants in the Context of Transnational Biomedical Research: From Conceptual Considerations to Practical Implications.
Outsourcing clinical trials sponsored by pharmaceutical companies from industrialized countries to low- (middle)-income countries - summarized as transnational biomedical research (TBR) - has lead to many concerns about ethical standards. Whether study participants are particularly vulnerable is one of those concerns. However, the concept of vulnerability is still vague and varies in its definition. Despite the fact that important international ethical guidelines such as the Declaration of Helsinki by the World Medical Association or the Ethical Guidelines for Biomedical Research Involving Human Subjects by the Council of International Organizations of Medical Sciences refer to vulnerability as ethical principle, each of their approaches are different. To overcome these shortcomings, we analyze and unite different approaches of vulnerability and develop practical criteria in order to operationalize the concept especially for the context of TBR. These criteria refer to the context of a study as well as the characteristics and the current living situation of study participants. Based on a case study of an HIV-vaccine-trial conducted in India we demonstrate how those criteria can be applied in a retrospective way to identify potential ethical conflicts. The criteria can also indicate a prospective function for ethical pre-assessment. For this, we provide an outlook for three major topics: 1. Vulnerability as a normative concept: Different ways of protection; 2. The relevance of transparency and 3. Vulnerability as an instrument to increase decision participation of human subjects.


Does the choice of bottle nipple affect the oral feeding performance of very-low-birthweight (VLBW) infants?
There is a continuous debate regarding the best bottle nipple to be used to enhance the bottle-feeding performance of a preterm infant. To verify that feeding performance can be improved by using the bottle nipple with the physical characteristics that enhance infants' sucking skills. Ten "healthy" VLBW infants (941+/-273 g) were recruited. Feeding performance was monitored at two time periods, when taking 1-2 and 6-8 oral feedings/d. At each time and within 24 h, performance was monitored using three different bottle nipples offered in a randomized order. Rate of milk transfer (ml/min) was the primary outcome measure. The sucking skills monitored comprised stage of sucking, suction amplitude, and duration of the generated negative intraoral suction pressure. At both times, infants demonstrated a similar rate of milk transfer among all three nipples. However, the stage of sucking, suction amplitude, and duration of the generated suction were significantly different between nipples at 1-2, but not 6-8 oral feedings/d. We did not identify a particular bottle nipple that enhanced bottle feeding in healthy VLBW infants. Based on the notion that afferent sensory feedback may allow infants to adapt to changing conditions, we speculate that infants can modify their sucking skills in order to maintain a rate of milk transfer that is appropriate with the level of suck-swallow-breathe coordination achieved at a particular time. Therefore, it is proposed that caretakers should be more concerned over monitoring the coordination of suck-swallow-breathe than over the selection of bottle nipples.


Low frequency of treatment of osteoporosis among postmenopausal women following a fracture.
Osteoporosis is a major cause of morbidity. Treatment of osteoporosis reduces the risk of fracture, particularly for postmenopausal women with a history of fracture. A retrospective study was conducted using the automated databases of 7 health maintenance organizations to evaluate the use of drugs recommended for secondary prevention of osteoporotic fracture. Women 60 years and older with an inpatient or outpatient diagnostic code for a fracture of the hip, vertebra, or wrist between October 1, 1994, and September 30, 1996, and at least 1 year of continuous enrollment with a drug benefit plan following the date of fracture, were identified. The frequency of use of medications for the treatment of osteoporosis (estrogen replacement therapy, bisphosphonates, and calcitonin) during the 1-year period following the date of the initial fracture was estimated overall and according to patient age, fracture site, and year of fracture. During the study period, 3492 women 60 years and older were diagnosed with a fracture of the hip, vertebra, or wrist, and met the inclusion criteria. Of these patients, 822 (24%) received a drug for osteoporosis treatment during the year following the fracture. The proportion of women receiving treatment for osteoporosis was approximately 2-fold higher among those with a fracture of the vertebra (44%) than among those with a fracture of the hip (21%) or wrist (23%) (P<.001). Of the 2605 women who had not been treated for osteoporosis in the 90 days before a fracture, 14% received treatment for osteoporosis in the year following a fracture. Increasing age was associated with a reduced likelihood of receiving osteoporosis treatment (P<.001). Most of the older women who had experienced a fracture of the hip, vertebra, or wrist did not receive drug treatment for osteoporosis within 1 year following the fracture. Interventions to improve the detection and treatment of osteoporosis in high-risk patients need to be developed.


Peripheral Nerve Block as a Supplement to Light or Deep General Anesthesia in Elderly Patients Receiving Total Hip Arthroplasty: A Prospective Randomized Study.
Peripheral nerve block combined with general anesthesia is a preferable anesthesia method for elderly patients receiving hip arthroplasty. The depth of sedation may influence patient recovery. Therefore, we investigated the influence of peripheral nerve blockade and different intraoperative sedation levels on the short-term recovery of elderly patients receiving total hip arthroplasty. Patients aged 65 years and older undergoing total hip arthroplasty were randomized into 3 groups: a general anesthesia without lumbosacral plexus block group, and 2 general anesthesia plus lumbosacral plexus block groups, each with a different level of sedation (light or deep). The extubation time and intraoperative consumption of propofol, sufentanil, and vasoactive agent were recorded. Postoperative delirium and early postoperative cognitive dysfunction were assessed using the Confusion Assessment Method and Mini-Mental State Examination, respectively. Postoperative analgesia was assessed by the consumption of patient-controlled analgesics and visual analog scale scores. Discharge time and complications over a 30-day period were also recorded. Lumbosacral plexus block reduced opioid intake. With lumbosacral plexus block, intraoperative deep sedation was associated with greater intake of propofol and vasoactive agent. In contrast, patients with lumbosacral plexus block and intraoperative light sedation had lower incidences of postoperative delirium and postoperative cognitive decline, and earlier discharge readiness times. The 3 groups showed no difference in complications within 30 days of surgery. Lumbosacral plexus block reduced the need for opioids and offered satisfactory postoperative analgesia. It led to better postoperative outcomes in combination with intraoperative light sedation (high bispectral index).


The nucleotide sequence and genomic organization of Citrus leaf blotch virus: candidate type species for a new virus genus.
The complete nucleotide sequence of Citrus leaf blotch virus (CLBV) was determined. CLBV genomic RNA (gRNA) has 8747 nt, excluding the 3'-terminal poly(A) tail, and contains three open reading frames (ORFs) and untranslated regions (UTR) of 73 and 541 nucleotides at the 5' and 3' termini, respectively. ORF1 potentially encodes a 227.4-kDa polypeptide, which has methyltransferase, papain-like protease, helicase, and RNA-dependent RNA polymerase motifs. ORF2 encodes a 40.2-kDa polypeptide containing a motif characteristic of cell-to-cell movement proteins. The 40.7-kDa polypeptide encoded by ORF3 was identified as the coat protein. The genome organization of CLBV resembles that of viruses in the genus Trichovirus, but they differ in various aspects: (i) in trichoviruses ORF2 overlaps ORFs 1 and 3, whereas in CLBV, ORFs 2 and 3 are separated and ORFs 1 and 2 overlap in one nucleotide; (ii) CLBV gRNA and CP are larger than those of trichoviruses; and (iii) the CLBV 3' UTR is larger than that of trichoviruses. Phylogenetic comparisons based on CP amino acid signatures clearly separates CLBV from trichoviruses. Also contrasting with trichoviruses, CLBV could not be transmitted to Chenopodium quinoa Willd. Considering these singularities, we propose that CLBV should be included in a new virus genus.


Biological sensing of small field differences by magnetically sensitive chemical reactions.
There is evidence that animals can detect small changes in the Earth's magnetic field by two distinct mechanisms, one using the mineral magnetite as the primary sensor and one using magnetically sensitive chemical reactions. Magnetite responds by physically twisting, or even reorienting the whole organism in the case of some bacteria, but the magnetic dipoles of individual molecules are too small to respond in the same way. Here we assess whether reactions whose rates are affected by the orientation of reactants in magnetic fields could form the basis of a biological compass. We use a general model, incorporating biological components and design criteria, to calculate realistic constraints for such a compass. This model compares a chemical signal produced owing to magnetic field effects with stochastic noise and with changes due to physiological temperature variation. Our analysis shows that a chemically based biological compass is feasible with its size, for any given detection limit, being dependent on the magnetic sensitivity of the rate constant of the chemical reaction.


4F2hc stabilizes GLUT1 protein and increases glucose transport activity.
Glucose transporter 1 (GLUT1) is widely distributed throughout various tissues and contributes to insulin-independent basal glucose uptake. Using a split-ubiquitin membrane yeast two-hybrid system, we newly identified 4F2 heavy chain (4F2hc) as a membrane protein interacting with GLUT1. Though 4F2hc reportedly forms heterodimeric complexes between amino acid transporters, such as LAT1 and LAT2, and regulates amino acid uptake, we investigated the effects of 4F2hc on GLUT1 expression and the associated glucose uptake. First, FLAG-tagged 4F2hc and hemagglutinin-tagged GLUT1 were overexpressed in human embryonic kidney 293 cells and their association was confirmed by coimmunoprecipitation. The green fluorescent protein-tagged 4F2hc and DsRed-tagged GLUT1 showed significant, but incomplete, colocalization at the plasma membrane. In addition, an endogenous association between GLUT1 and 4F2hc was demonstrated using mouse brain tissue and HeLa cells. Interestingly, overexpression of 4F2hc increased the amount of GLUT1 protein in HeLa and HepG2 cells with increased glucose uptake. In contrast, small interfering RNA (siRNA)-mediated 4F2hc gene suppression markedly reduced GLUT1 protein in both cell types, with reduced glucose uptake. While GLUT1 mRNA levels were not affected by overexpression or gene silencing of 4F2hc, GLUT1 degradation after the addition of cycloheximide was significantly suppressed by 4F2hc overexpression and increased by 4F2hc siRNA treatment. Taken together, these observations indicate that 4F2hc is likely to be involved in GLUT1 stabilization and to contribute to the regulation of not only amino acid but also glucose metabolism.


Texton-based segmentation of retinal vessels.
With improvements in fundus imaging technology and the increasing use of digital images in screening and diagnosis, the issue of automated analysis of retinal images is gaining more serious attention. We consider the problem of retinal vessel segmentation, a key issue in automated analysis of digital fundus images. We propose a texture-based vessel segmentation algorithm based on the notion of textons. Using a weak statistical learning approach, we construct textons for retinal vasculature by designing filters that are specifically tuned to the structural and photometric properties of retinal vessels. We evaluate the performance of the proposed approach using a standard database of retinal images. On the DRIVE data set, the proposed method produced an average performance of 0.9568 specificity at 0.7346 sensitivity. This compares well with the best-published results on the data set 0.9773 specificity at 0.7194 sensitivity [Proc. SPIE5370, 648 (2004)].


On the evaporation kinetics of [60] fullerene in aromatic organic solvents.
We investigate the effect of C60 fullerene nanospheres on the evaporation kinetics of a number of aromatic solvents with different levels of molecular association, namely, benzene, toluene, and chlorobenzene. The dependence of the evaporation rate on the fullerene concentration is not monotonic but rather exhibits maxima and minima. The results strongly support the notion of molecular structuring within the liquid solvent controlled by the nature of the fullerene/solvent interaction and the level of molecular association within the solvent itself.


Genomics, evolution, and crystal structure of a new family of bacterial spore kinases.
Bacterial spore formation is a complex process of fundamental relevance to biology and human disease. The spore coat structure is complex and poorly understood, and the roles of many of the protein components remain unclear. We describe a new family of spore coat proteins, the bacterial spore kinases (BSKs), and the first crystal structure of a BSK, YtaA (CotI) from Bacillus subtilis. BSKs are widely distributed in spore-forming Bacillus and Clostridium species, and have a dynamic evolutionary history. Sequence and structure analyses indicate that the BSKs are CAKs, a prevalent group of small molecule kinases in bacteria that is distantly related to the eukaryotic protein kinases. YtaA has substantial structural similarity to CAKs, but also displays distinctive features that broaden our understanding of the CAK group. Evolutionary constraint analysis of the protein surfaces indicates that members of the BSK family have distinct clade-conserved patterns in the substrate binding region, and probably bind and phosphorylate distinct targets. Several classes of BSKs have apparently independently lost catalytic activity to become pseudokinases, indicating that the family also has a major noncatalytic function.


Patient preference and willingness to pay for transient elastography versus liver biopsy: A perspective from British Columbia.
The cost of liver biopsy (LB) is publicly funded in British Columbia, while the cost of transient elastography (FibroScan [FS], Echosens, France) is not. Consequently, there is regional variation regarding FS access and monitoring of liver disease progression. To evaluate patient preference for FS versus LB and to assess the willingness to self-pay for FS. Questionnaires were distributed in clinic and via mail to LB-experienced and LB-naive patients who underwent FS at Vancouver General Hospital, Vancouver, British Columbia. The overall response rate was 76%. Of the 422 respondents, 205 were LB-experienced. The mean age was 53.5 years, 50.2% were male, 54.7% were Caucasian, 38.2% had hepatitis C and 26.3% had an annual household income >$75,000. Overall, 95.4% of patients preferred FS to LB. FS was associated with greater comfort than LB, with the majority reporting no discomfort during FS (84.1% versus 7.8% for LB), no discomfort after (96.2% versus 14.6% LB) and no feelings of anxiety after FS explanation (78.2% versus 12.7% LB). FS was also associated with greater speed, with the majority reporting short test duration (97.2% versus 48.3% LB) and short wait for the test result (95.5% versus 30.2% LB). Most (75.3%) respondents were willing to self-pay for FS, with 26.3% willing to pay $25 to $49. Patients with unknown liver disease preferred LB (OR [FS preference] 0.20 [95% CI 0.07 to 0.53]). FS was the preferred method of assessing liver fibrosis among patients, with the majority willing to self-pay. To ensure consistency in access, provincial funding for FS is needed. However, LB remains the procedure of choice for individuals with an unknown diagnosis.


Designing prospective cohort studies for assessing reproductive and developmental toxicity during sensitive windows of human reproduction and development--the LIFE Study.
The relationship between the environment and human fecundity and fertility remains virtually unstudied from a couple-based perspective in which longitudinal exposure data and biospecimens are captured across sensitive windows. In response, we completed the LIFE Study with methodology that intended to empirically evaluate a priori purported methodological challenges: implementation of population-based sampling frameworks suitable for recruiting couples planning pregnancy; obtaining environmental data across sensitive windows of reproduction and development; home-based biospecimen collection; and development of a data management system for hierarchical exposome data. We used two sampling frameworks (i.e., fish/wildlife licence registry and a direct marketing database) for 16 targeted counties with presumed environmental exposures to persistent organochlorine chemicals to recruit 501 couples planning pregnancies for prospective longitudinal follow-up while trying to conceive and throughout pregnancy. Enrolment rates varied from <1% of the targeted population (n = 424,423) to 42% of eligible couples who were successfully screened; 84% of the targeted population could not be reached, while 36% refused screening. Among enrolled couples, ∼ 85% completed daily journals while trying; 82% of pregnant women completed daily early pregnancy journals, and 80% completed monthly pregnancy journals. All couples provided baseline blood/urine samples; 94% of men provided one or more semen samples and 98% of women provided one or more saliva samples. Women successfully used urinary fertility monitors for identifying ovulation and home pregnancy test kits. Couples can be recruited for preconception cohorts and will comply with intensive data collection across sensitive windows. However, appropriately sized sampling frameworks are critical, given the small percentage of couples contacted found eligible and reportedly planning pregnancy at any point in time.


Engineered liposomes sequester bacterial exotoxins and protect from severe invasive infections in mice.
Gram-positive bacterial pathogens that secrete cytotoxic pore-forming toxins, such as Staphylococcus aureus and Streptococcus pneumoniae, cause a substantial burden of disease. Inspired by the principles that govern natural toxin-host interactions, we have engineered artificial liposomes that are tailored to effectively compete with host cells for toxin binding. Liposome-bound toxins are unable to lyse mammalian cells in vitro. We use these artificial liposomes as decoy targets to sequester bacterial toxins that are produced during active infection in vivo. Administration of artificial liposomes within 10 h after infection rescues mice from septicemia caused by S. aureus and S. pneumoniae, whereas untreated mice die within 24-33 h. Furthermore, liposomes protect mice against invasive pneumococcal pneumonia. Composed exclusively of naturally occurring lipids, tailored liposomes are not bactericidal and could be used therapeutically either alone or in conjunction with antibiotics to combat bacterial infections and to minimize toxin-induced tissue damage that occurs during bacterial clearance.


ACPA IgG galactosylation associates with disease activity in pregnant patients with rheumatoid arthritis.
Patients with autoantibody-positive rheumatoid arthritis (RA) are less likely to experience pregnancy-induced improvement of RA disease activity (DAS28-C reactive protein (CRP)) compared with patients with autoantibody-negative RA. Anti-citrullinated protein antibodies (ACPAs) are the most specific autoantibodies for RA. We previously demonstrated that disease improvement is associated with changes in total IgG glycosylation, which regulate antibody effector function. Therefore, we sought to analyse the ACPA-IgG glycosylation profile during pregnancy with the aim to understand the lower change of pregnancy-induced improvement of the disease in patients with autoantibody-positive RA. ACPA-IgGs were purified from ACPA-positive patient sera (n=112) of the Pregnancy-induced Amelioration of Rheumatoid Arthritis cohort, a prospective study designed to investigate pregnancy-associated improvement of RA. The fragment crystallisable (Fc)glycosylation profile of ACPA-IgGs was characterised by mass spectrometry and compared with that of total IgG derived from the same patients or from ACPA-negative patients. All ACPA-IgG subclasses display significant changes in the level of galactosylation and sialylation during pregnancy, although less pronounced than in total IgG. The pregnancy-induced increase in ACPA-IgG galactosylation, but not sialylation, associates with lower DAS28-CRP. In ACPA-positive patients, no such association was found with changes in the galactosylation of total IgG, whereas in ACPA-negative patients changes in disease activity correlated well with changes in the galactosylation of total IgG. In ACPA-positive RA, the pregnancy-induced change in galactosylation of ACPA-IgG, and not that of total IgG, associates with changes in disease activity. These data may indicate that in ACPA-positive patients the galactosylation of ACPA-IgG is of more pathogenic relevance than that of total IgG.


Assisted Ventilation in Patients with Acute Respiratory Distress Syndrome: Lung-distending Pressure and Patient-Ventilator Interaction.
In patients with acute respiratory distress syndrome (ARDS), the use of assisted mechanical ventilation is a subject of debate. Assisted ventilation has benefits over controlled ventilation, such as preserved diaphragm function and improved oxygenation. Therefore, higher level of "patient control" of ventilator assist may be preferable in ARDS. However, assisted modes may also increase the risk of high tidal volumes and lung-distending pressures. The current study aims to quantify how differences in freedom to control the ventilator affect lung-protective ventilation, breathing pattern variability, and patient-ventilator interaction. Twelve patients with ARDS were ventilated in a randomized order with assist pressure control ventilation (PCV), pressure support ventilation (PSV), and neurally adjusted ventilatory assist (NAVA). Transpulmonary pressure, tidal volume, diaphragm electrical activity, and patient-ventilator interaction were measured. Respiratory variability was assessed using the coefficient of variation of tidal volume. During inspiration, transpulmonary pressure was slightly lower with NAVA (10.3 ± 0.7, 11.2 ± 0.7, and 9.4 ± 0.7 cm H2O for PCV, PSV, and NAVA, respectively; P < 0.01). Tidal volume was similar between modes (6.6 [5.7 to 7.0], 6.4 [5.8 to 7.0], and 6.0 [5.6 to 7.3] ml/kg for PCV, PSV, and NAVA, respectively), but respiratory variability was higher with NAVA (8.0 [6.4 to 10.0], 7.1 [5.9 to 9.0], and 17.0 [12.0 to 36.1] % for PCV, PSV, and NAVA, respectively; P < 0.001). Patient-ventilator interaction improved with NAVA (6 [5 to 8] % error) compared with PCV (29 [14 to 52] % error) and PSV (12 [9 to 27] % error); P < 0.0001. In patients with mild-to-moderate ARDS, increasing freedom to control the ventilator maintains lung-protective ventilation in terms of tidal volume and lung-distending pressure, but it improves patient-ventilator interaction and preserves respiratory variability.