Routine brief risk-reduction counseling with biannual STD testing reduces STD incidence among HIV-infected men who have sex with men in care.
We evaluated whether routine biannual sexually transmitted disease (STD) testing coupled with brief risk-reduction counseling reduces STD incidence and high-risk behaviors. The SUN study is a prospective observational HIV cohort study conducted in 4 US cities. At enrollment and every 6 months thereafter, participants completed a behavioral survey and were screened for STDs, and if diagnosed, were treated. Medical providers conducted brief risk-reduction counseling with all patients. Among men who have sex with men (MSM), we examined trends in STD incidence and rates of self-reported risk behaviors before and after exposure to the risk-reduction intervention. The "preintervention" visit was the study visit that was at least 6 months after enrollment STD screening and treatment and at which the participant was first exposed to the intervention. The "postintervention" visit was 12 months later. Among 216 MSM with complete STD and behavioral data, median age was 44.5 years; 77% were non-Hispanic white; 83% were on highly active antiretroviral treatment; 84% had an HIV RNA level <400 copies/mL and the median CD4 (cluster of differentiation 4) count was 511 cells/mm. Twelve months after first exposure to the risk-reduction intervention, STD incidence declined from 8.8% to 4.2% (P = 0.041). Rates of unprotected receptive or insertive anal intercourse with HIV-positive partners increased (19% to 25%, P = 0.024), but did not change with HIV-negative partners or partners of unknown HIV status (24% to 22%, P = 0.590). STD incidence declined significantly among HIV-infected MSM after implementing frequent, routine STD testing coupled with risk-reduction counseling. These findings support adoption of routine STD screening and risk-reduction counseling for HIV-infected MSM.


Molecular and biological characterization of Hammondia heydorni-like oocysts from a dog fed hearts from naturally infected white-tailed deer (Odocoileus Virginianus).
Neospora caninum and Hammondia heydorni are morphologically and phylogenetically related coccidians that are found in dogs. Although there is serological evidence of N. caninum infection in the white-tailed deer (Odocoileus virginianus), the parasite has not been yet isolated from the tissues of this host. In an attempt to isolate N. caninum from deer, hearts from 4 deer with antibodies to N. caninum were fed to 2 dogs. One of these dogs shed unsporulated oocysts 12-14 microm in diameter. Sporulated oocysts were not infective to Mongolian gerbils (Meriones ungulatus), and DNA isolated from these oocysts was not amplified using N. caninum-specific primers. However, positive amplification with the H. heydorni-specific first internal transcribed spacer (ITS-1) primers and common toxoplasmatiid ITS-1 primers confirmed the presence of H. heydorni DNA in the samples. The oocysts were considered to be H. heydorni on the basis of their morphology, biology, and molecular characteristics. This is the first record of a H. heydorni-like parasite in the white-tailed deer.


Endogenous Pain Facilitation Rather Than Inhibition Differs Between People with Chronic Fatigue Syndrome, Multiple Sclerosis, and Controls: An Observational Study.
Commonalities in the core symptoms of fatigue and cognitive dysfunction experienced by chronic fatigue syndrome (CFS, also known as ME) and multiple sclerosis (MS) patients have been described. Many CFS and MS patients also experience chronic pain, which has been attributed to central sensitization in both groups of patients. However, the characteristics of pain in CFS and MS patients have not been compared. To compare experimental pain measurements in patients with CFS or MS and healthy controls. Observational study. This study took place in Belgium at Vrije Universiteit Brussel and the University of Antwerp. Pressure pain thresholds, temporal summation, conditioned pain modulation, and occlusion cuff pressure thresholds rated as painful (1st cuff pressure threshold) and as 3/10 on a verbal numerical scale (2nd cuff pressure threshold) were measured in patients with CFS (n = 48), MS (n = 19) and healthy pain-free controls (n = 30). Adjusted between-group differences were estimated using linear regression models. Finger pain pressure thresholds of patients with CFS, compared with patients with MS, were 25% lower (difference ratio 0.75 [95% CI 0.59, 0.95], P = 0.02) and shoulder pain pressure thresholds were 26% lower (difference ratio 0.74 [0.52, 1.04], P = 0.08). Compared with patients with MS, patients with CFS had 29% lower first cuff pressure threshold (difference ratio 0.71 [0.53, 0.94], P = 0.02) and 41% lower 2nd cuff pressure threshold (0.59 [0.41, 0.86], P = 0.006). Finger temporal summation was higher in patients with CFS than in patients with MS (mean difference 1.15 [0.33, 1.97], P = 0.006), but there were no differences in shoulder temporal summation or conditioned pain modulation at either site. Differences between patients with CFS and MS tended to be greater than between either patient group and healthy controls. Pain pressure thresholds and cuff pressure thresholds tended to be positively correlated, and temporal summation negatively correlated, with higher physical function and lower fatigue in both groups of patients. Subjective pain in patients with CFS but not in patients with MS was strongly negatively correlated with pain pressure thresholds and cuff pressure thresholds, and positively correlated with temporal summation. The main limitations of our study are the relatively small sample sizes, its cross-sectional design, and its exploratory nature. We found differences in the characteristics of pain symptoms reported by patients with CFS and patients with MS, which suggest different underlying mechanisms. Specifically, overactive endogenous pain facilitation was characteristic of pain in patients with CFS but not in patients with MS, suggesting a greater role for central sensitization in CFS.


Characteristics of Human Endometrial Stem Cells in Tissue and Isolated Cultured Cells: An Immunohistochemical Aspect.
The aim of this study was to investigate the percentage of the stem cells population in human endometrial tissue sections and cultured cells at fourth passage. Human endometrial specimens were divided into two parts, one part for morphological studies and the other part for in vitro culture. Full thickness of human normal endometrial sections and cultured endometrial cells at fourth passage were analyzed via immunohistochemistry for CD146 and some stemness markers such as Oct4, Nanog, Sox2, and Klf4 and the expression of typical mesenchymal stem cell markers CD90, CD105. 11.88 ± 1.29% of human endometrial cells whitin tissue sections expressed CD146 marker vs. 28±2.3% of cultured cells, CD90 and CD105 were expressed by functionalis stroma (85±2.4 and 89±3.2%) than basalis stroma (16±1.4 and 17±1.9%), respectively (P<0.05). Oct4 and Nanog-expressing cells comprise 1.43±0.08 and 0.54±0.01% of endometrial stromal cells in endometrial sections vs. 12±3.1% and 8±2.9% of cultured cells, respectively. They reside near the glands in the basal layer of endometrium. Sox2 and Klf4 were not commonly expressed in tissue samples and cultured cells. CD9 and EpCAM were expressed by epithelial cells of the endometrium, rather than by stroma or perivascular cells. The human endometrial stem cells and pluripotency markers may be localized more in basalis layer of endometrium. The immunostaining observations of endometrial cells at fourth passage were correlated with the immunohistochemistry data.


Curcumin pretreatment attenuates inflammation and mitochondrial dysfunction in experimental stroke: The possible role of Sirt1 signaling.
The effects of curcumin (CCM) on cerebral ischemia/reperfusion injury are not well understood. The aim of this study was to investigate whether CCM attenuates inflammation and mitochondrial dysfunction in a rat model of cerebral ischemia/reperfusion injury and whether Sirt1 is involved in these potential protective effects. Sirtinol, a Sirt1 inhibitor, was used to elucidate the underlying mechanism. Rats were subjected to 2h of transient middle cerebral artery occlusion (MCAO), followed by reperfusion for 24h. Brain magnetic resonance imaging (MRI) was used to detect infarct volumes. Neurological scores and brain water content were also assessed. Levels of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in the brain were detected using commercial enzyme-linked immunosorbent assay (ELISA) kits. Expression of SIRT1, acetylated p53 (Ac-p53), Bcl-2, and Bax was measured by western blotting. Our results suggested that CCM exerted a neuroprotective effect, as shown by reduced infarct volumes and brain edema and improved neurological scores. CCM also exerted anti-inflammatory effects, as indicated by decreased TNF-α and IL-6 levels in the brain. CCM elevated mitochondrial membrane potential, mitochondrial complex I activity, and mitochondrial cytochrome c levels, but reduced cytosolic cytochrome c levels. Moreover, CCM upregulated SIRT1 and Bcl-2 expression and downregulated Ac-p53 and Bax expression. These effects of CCM were abolished by sirtinol. In conclusion, our results demonstrate that CCM treatment attenuates ischemic stroke-induced brain injury via activation of SIRT1.


Health care professionals' personal and professional views of herbal medicines in the United Kingdom.
Health care professionals (HCPs) have a pivotal role in optimizing patient care and should be familiar with complementary and alternative medicines. The aim of the study was to explore UK-based HCP personal and professional opinions and experiences of herbal medicines (HMs). An online questionnaire was distributed via social media to recruit (n = 112) a range of HCPs from across the United Kingdom. HCPs from primary and secondary care, the private sector, and academia took part. A large proportion of participants (62%) said they did not personally use any HMs, and 38% did use HMs. HCPs who had personally used HMs had a positive impression of HMs and were more likely to recommend HMs to patients than those who had not used HMs themselves. Participants were given the opportunity to share their perceptions on the safety and efficacy of HMs and their experiences with patients reporting adverse drug reactions to HMs and herb-drug interactions. HCPs identified their lack of knowledge on HMs and insufficient training, which made them unable to advise patients on the safe use of HMs. More education on HMs would help improve HCP knowledge of HMs and help them make better informed decisions when considering patient pharmaceutical care plans.


Consensus recommendations for the management of constipation in patients with advanced, progressive illness.
Constipation is a highly prevalent and distressing symptom in patients with advanced, progressive illnesses. Although opioids are one of the most common causes of constipation in patients with advanced, progressive illness, it is important to note that there are many other potential etiologies and combinations of causes that should be taken into consideration when making treatment decisions. Management approaches involve a combination of good assessment techniques, preventive regimens, appropriate pharmacological treatment of established constipation, and frequent monitoring. In this vulnerable patient population, maintenance of comfort and respect for individual preferences and sensitivities should be overriding considerations when making clinical decisions. This consensus document was developed by a multidisciplinary group of leading Canadian palliative care specialists in an effort to define best practices in palliative constipation management that will be relevant and useful to health care professionals. Although a wide range of options exists to help treat constipation and prevent its development or recurrence, there is a limited body of evidence evaluating pharmacological interventions. These recommendations are, therefore, based on the best of the available evidence, combined with expert opinion derived from experience in clinical practice. This underscores the need for further clinical evaluation of the available agents to create a robust, evidence-based foundation for treatment decisions in the management of constipation in patients with advanced, progressive illness.


Dissolution DNP using trityl radicals at 7 T field.
Dissolution DNP has become an important method to generate highly polarized substrates such as pyruvic acid for in vivo imaging and localized spectroscopy. In a quest to further increase the polarization levels, which is important for in vivo MRI employing 13C detection, we describe the design and implementation of a new DNP polarizer that is suitable for dissolution operation at 7 T static magnetic field and a temperature of 1.4 K. We describe all important sample preparation steps and experimental details necessary to optimize trityl based samples for use in our polarizer at this higher field. In [1-13C]-pyruvic acid polarization levels of about 56% are achieved, compared to typical polarization levels of about 35-45% at a standard field of 3.4 T. At the same time, the polarization build-up time increases significantly from about 670 s at 3.4 T to around 1300-1900 s at 7 T, depending on the trityl derivate used. We also investigate the effect of adding trace amounts of Gd3+ to the samples. While one trityl compound does not exhibit any benefit, the other profits significantly, boosting achievable polarization by 6%.


[The Theory and Trend of Microhistory: History of Medicine].
Microhistory, first developed in the 1970s, is the study of the past on a very small scale applying zoom-in methodology. Although microhistory had been introduced to Korea during the late 1990s, there still exists much of misunderstanding and confusions surrounding its nature. Microhistory is to be distinguished from the monographs which deal with petty subject, or from the history of everyday life, and from the case studies. In the field of the history of medicine, there are not many microhistories proper. Several works that claim to utilize microhistorical approach, could not be categorized as microhistory because they carry strong characteristics of macrohistory, specifically those of the disciplinary history or the case studies. The well known work of Harold J. Cook, Trials of an Ordinary Doctor, is not an exception. These studies fail to materialize the critical mind of microhistory that pursues to write a history from below and to restore the agency of obscure people. However, Guido Ruggiero's "The Strange Death of Margarita Marcellini," David Cressy's Travesties and Transgressions in Tudor and Stuart England, and Laurel Thatcher Ulrich's A Midwife's Tale clearly demonstrate the attributes, characteristics, and methodologies of microhistory. These studies well display the emphasis of microhistory, which reveal the complexity of early modern medicine, and the complicated function of individual relationships within each and every social setting. Recently, some scholars begin to suggest that the rigid definition of microhistory should be softened, arguing that there could be various types of microhistory. The history of medicine has many advantage of aptly applying many virtues of microhistory: the de-territoriality of diseases, the peculiar elements of the training and practice in hospitals and medical schools which call for anthropological survey, and the possibility of utilizing doctor's records that contain the confessions of the patients. Also, medical historians need to expand the scope of medical provider for their analysis, incorporating pseudo-medical doctor. The essential property of microhistory should be found in its request for various new subjects and fresh perspectives.


Small and ugly? Phylogenetic analyses of the "selfing syndrome" reveal complex evolutionary fates of monomorphic primrose flowers.
One of the most common trends in plant evolution, loss of self-incompatibility and ensuing increases in selfing, is generally assumed to be associated with a suite of phenotypic changes, notably a reduction of floral size, termed the selfing syndrome. We investigate whether floral morphological traits indeed decrease in a deterministic fashion after losses of self-incompatibility, as traditionally expected, using a phylogeny of 124 primrose species containing nine independent transitions from heterostyly (heteromorphic incompatibility) to homostyly (monomorphic self-compatibility), a classic system for evolution of selfing. We find similar overall variability of homostylous and heterostylous species, except for diminished herkogamy in homostyles. Bayesian mixed models demonstrate differences between homostylous and heterostylous species in all traits, but net effects across species are small (except herkogamy) and directionality differs among traits. Strongly drift-like evolutionary trajectories of corolla tube length and corolla diameter inferred by Ornstein-Uhlenbeck models contrast with expected deterministic trajectories toward small floral size. Lineage-specific population genetic effects associated with evolution of selfing may explain that reductions of floral size represent one of several possible outcomes of floral evolution after loss of heterostyly in primroses. Contrary to the traditional paradigm, selfing syndromes may, but do not necessarily evolve in response to increased selfing.


Spatially explicit estimates of prey consumption reveal a new krill predator in the Southern Ocean.
Development in foraging behaviour and dietary intake of many vertebrates are age-structured. Differences in feeding ecology may correlate with ontogenetic shifts in dispersal patterns, and therefore affect foraging habitat and resource utilization. Such life-history traits have important implications in interpreting tropho-dynamic linkages. Stable isotope ratios in the whiskers of sub-yearling southern elephant seals (Mirounga leonina; n = 12) were used, in conjunction with satellite telemetry and environmental data, to examine their foraging habitat and diet during their first foraging migration. The trophic position of seals from Macquarie Island (54°30'S, 158°57'E) was estimated using stable carbon (δ(1) (3)C) and nitrogen (δ(15)N) ratios along the length of the whisker, which provided a temporal record of prey intake. Satellite-relayed data loggers provided details on seal movement patterns, which were related to isotopic concentrations along the whisker. Animals fed in waters south of the Polar Front (>60°S) or within Commission for the Conservation of Antarctic Marine Living Resources (CCAMLR) Statistical Subareas 88.1 and 88.2, as indicated by both their depleted δ(1) (3)C (<-20‰) values, and tracking data. They predominantly exploited varying proportions of mesopelagic fish and squid, and crustaceans, such as euphausiids, which have not been reported as a prey item for this species. Comparison of isotopic data between sub-yearlings, and 1, 2 and 3 yr olds indicated that sub-yearlings, limited by their size, dive capabilities and prey capture skills to feeding higher in the water column, fed at a lower trophic level than older seals. This is consistent with the consumption of euphausiids and most probably, Antarctic krill (Euphausia superba), which constitute an abundant, easily accessible source of prey in water masses used by this age class of seals. Isotopic assessment and concurrent tracking of seals are successfully used here to identify ontogenetic shifts in broad-scale foraging habitat use and diet preferences in a highly migratory predator.


Indonesian healthcare professionals' experiences in rural and urban settings during the first wave of COVID-19: A qualitative study.
During the COVID-19 pandemic, healthcare workers (HCWs) faced unprecedented challenges, increased workload, and often struggled to provide healthcare services. We explored the experiences faced by HCWs working at primary healthcare centers (PHCs) and hospitals across urban and rural settings in Indonesia. As part of a larger multi-country study, we conducted semi-structured in-depth interviews with a purposive sample of Indonesian HCWs. We used thematic analysis to identify the main challenges described by the participants. We interviewed 40 HCWs between December 2020 and March 2021. We identified that challenges varied depending on their role. i) For those in clinical roles, challenges included maintaining trust with communities, and patient referral issues; ii) for those in non-clinical roles, sub-optimal laboratory capacity and logistics, and lack of training were the main challenges; iii) for managerial roles, challenges included access to budget and supplies, and staff shortages due to isolation and overwork. There were also several cross-cutting challenges across all the roles including limited or rapidly changing information (in urban settings), and culture and communication (in rural settings). All of these challenges contributed to mental health issues among all HCW cadres. HCWs across roles and settings were confronted with unprecedented challenges. Understanding the various challenges across different healthcare cadres and within different settings is crucial for supporting HCWs during pandemic times. In rural areas, in particular, HCWs should be more sensitive to cultural and linguistic differences to enhance the effectiveness and awareness of public health messages.


Female infants are more susceptible to the effects of maternal antenatal depression; findings from the Pelotas (Brazil) Birth Cohort Study.
We utilised data from the 2015 Pelotas Birth Cohort, a large prospective cohort in southern Brazil, to examine the association of moderate and severe antenatal depression with child birth outcomes and explore interactions with sociodemographic characteristics. Data was available for n = 3046 participants and their infants. We measured antenatal depression using the Edinburgh Postnatal Depression Scale (EPDS, ≥13 for moderate and ≥17 for severe depression). Outcome measures included gestational age, birth weight, length and head circumference, using the Intergrowth-21st standards. We controlled for known confounders including obstetric risk. We did not find differences in childbirth outcomes by maternal depression status for participants with at least moderate depression, although there was an increased risk for female offspring to be small for gestational age (SGA, OR 2.33[1.37,3.97]). For severe depression (EPDS≥17) we found an increased risk for lower APGAR scores (OR 1.63[1.02,2.60]) and being SGA (OR 1.77[1.06,2.97], with an increased risk for female offspring in particular to be in lower weight centiles (-10.71 [-16.83,-4.60]), to be SGA (OR 3.74[1.89, 7.44]) and in the lower 10th centile for length (OR 2.19[1.25,3.84]). include the use of a maternal report questionnaire to ascertain depressive symptoms. In this recent large longitudinal cohort in Brazil we did not find independent effects of depression on adverse birth outcomes or interactions with sociodemographic characteristics. We found an increased risk of being SGA for female offspring of women with moderate and severe depression, in line with other research suggesting females may be more susceptible to antenatal disturbances. This work was supported by the Wellcome Trust, United Kingdom (095582), the Brazilian National Research Council (CNPq) and the Coordination for the Improvement of Higher Education Personnel (CAPES). EN was supported by the UK Economic and Social Research Council GCRF Postdoctoral Fellowship (ES/P009794/1).


A Phase I proof-of-concept and safety trial of sildenafil to treat cerebral vasospasm following subarachnoid hemorrhage.
Studies show that phosphodiesterase-V (PDE-V) inhibition reduces cerebral vasospasm (CVS) and improves outcomes after experimental subarachnoid hemorrhage (SAH). This study was performed to investigate the safety and effect of sildenafil (an FDA-approved PDE-V inhibitor) on angiographic CVS in SAH patients. A2-phase, prospective, nonrandomized, human trial was implemented. Subarachnoid hemorrhage patients underwent angiography on Day 7 to assess for CVS. Those with CVS were given 10 mg of intravenous sildenafil in the first phase of the study and 30 mg in the second phase. In both, angiography was repeated 30 minutes after infusion. Safety was assessed by monitoring neurological examination findings and vital signs and for the development of adverse reactions. For angiographic assessment, in a blinded fashion, pre- and post-sildenafil images were graded as "improvement" or "no improvement" in CVS. Unblinded measurements were made between pre- and post-sildenafil angiograms. Twelve patients received sildenafil; 5 patients received 10 mg and 7 received 30 mg. There were no adverse reactions. There was no adverse effect on heart rate or intracranial pressure. Sildenafil resulted in a transient decline in mean arterial pressure, an average of 17% with a return to baseline in an average of 18 minutes. Eight patients (67%) were found to have a positive angiographic response to sildenafil, 3 (60%) in the low-dose group and 5 (71%) in the high-dose group. The largest degree of vessel dilation was an average of 0.8 mm (range 0-2.1 mm). This corresponded to an average percentage increase in vessel diameter of 62% (range 0%-200%). The results from this Phase I safety and proof-of-concept trial assessing the use of intravenous sildenafil in patients with CVS show that sildenafil is safe and well tolerated in the setting of SAH. Furthermore, the angiographic data suggest that sildenafil has a positive impact on human CVS.


Comparison of human solute carriers.
Solute carriers are eukaryotic membrane proteins that control the uptake and efflux of solutes, including essential cellular compounds, environmental toxins, and therapeutic drugs. Solute carriers can share similar structural features despite weak sequence similarities. Identification of sequence relationships among solute carriers is needed to enhance our ability to model individual carriers and to elucidate the molecular mechanisms of their substrate specificity and transport. Here, we describe a comprehensive comparison of solute carriers. We link the proteins using sensitive profile-profile alignments and two classification approaches, including similarity networks. The clusters are analyzed in view of substrate type, transport mode, organism conservation, and tissue specificity. Solute carrier families with similar substrates generally cluster together, despite exhibiting relatively weak sequence similarities. In contrast, some families cluster together with no apparent reason, revealing unexplored relationships. We demonstrate computationally and experimentally the functional overlap between representative members of these families. Finally, we identify four putative solute carriers in the human genome. The solute carriers include a biomedically important group of membrane proteins that is diverse in sequence and structure. The proposed classification of solute carriers, combined with experiment, reveals new relationships among the individual families and identifies new solute carriers. The classification scheme will inform future attempts directed at modeling the structures of the solute carriers, a prerequisite for describing the substrate specificities of the individual families.


Testing combined pharmacotherapies and behavioral interventions in alcohol dependence: rationale and methods.
Increasing knowledge about effective therapies for alcohol dependence calls for new research designs to examine treatment interactions between pharmacotherapies and behavioral interventions. In 1997, the National Institute on Alcohol Abuse and Alcoholism recruited 11 sites and a coordinating center for a large-scale (1,375 subjects), randomized placebo controlled trial to test 16 weeks of active treatment using naltrexone and acamprosate alone and in combination. Most participants receive 9 brief sessions delivered by medically trained providers to promote sobriety and enhance medication adherence (Medical Management, MM). Half the participants are also randomized to individualized psychotherapy (up to 20 sessions of Combined Behavioral Intervention, CBI), integrating elements of the successful behavioral interventions from Project MATCH. COMBINE seeks to evaluate the efficacy of the two most promising medications (naltrexone and acamprosate) both singly and together, when combined with different intensities of behavioral therapies. COMBINE incorporates a number of innovative design aspects, including a no-pill psychotherapy-alone condition, behavioral interventions that are both manual-guided and individualized, and pharmacotherapy dosing that is greater than in some previous trials. Two COMBINE pilot studies demonstrate the safety and acceptability of the combination pharmacotherapy dosing, and the feasibility of implementing the manualized behavioral interventions. This paper introduces COMBINE's goals, methods and analytic strategies, and their potential to improve multimodal treatment selection.


Phylogeny of Fomitopsis pinicola: a species complex.
Fungal species with a broad distribution may exhibit considerable genetic variation over their geographic ranges. Variation may develop among populations based on geographic isolation, lack of migration, and genetic drift, though this genetic variation may not always be evident when examining phenotypic characters. Fomitopsis pinicola is an abundant saprotrophic fungus found on decaying logs throughout temperate regions of the Northern Hemisphere. Phylogenetic studies have addressed the relationship of F. pinicola to other wood-rotting fungi, but pan-continental variation within F. pinicola has not been addressed using molecular data. While forms found growing on hardwood and softwood hosts exhibit variation in habit and appearance, it is unknown if these forms are genetically distinct. In this study, we generated DNA sequences of the nuc rDNA internal transcribed spacers (ITS), the TEF1 gene encoding translation elongation factor 1-α, and the RPB2 gene encoding the second largest subunit of RNA polymerase II for collections across all major geographic regions where this fungus occurs, with a primary focus on North America. We used Bayesian and maximum likelihood analyses and evaluated the gene trees within the species tree using coalescent methods to elucidate evolutionarily independent lineages. We find that F. pinicola sensu lato encompasses four well-supported, congruent clades: a European clade, southwestern US clade, and two sympatric northern North American clades. Each clade represents distinct species according to phylogenetic and population-genetic species concepts. Morphological data currently available for F. pinicola do not delimit these species, and three of the species are not specific to either hardwood or softwood trees. Originally described from Europe, F. pinicola appears to be restricted to Eurasia. Based on DNA data obtained from an isotype, one well-defined and widespread clade found only in North America represents the recently described Fomitopsis ochracea The remaining two North American clades represent previously undescribed species.


Effect of intermittent three-month treatment with different doses of Ukrain on subregional bone mineral density of the femur of ovariectomized rats.
Ukrain, thiophosphoric acid alkaloid derivative from Chelidonium majus L., was administered i.p. to ovariectomized rats in doses of 7, 14 and 28 mg/kg every other day for 10 days, followed by a 10-day break, and the procedure was performed five times. At the end of long-term treatment with Ukrain (24 h after the last dose of the drug) the rats' right femora were harvested and the bone densitometric parameters of the whole bone and distal metaphyseal and basicervical subregions were assessed using the dual-energy X-ray absorptiometry (DXA) densitometric method. The present results show a decrease in bone mineral density in groups of ovariectomized rats that received 7 mg/kg and 14 mg/kg of Ukrain versus untreated ovariectomized animals. Administration of Ukrain at a dose of 28 mg/kg did not significantly alter bone parameters of ovariectomized rats.


Regulation of the early expression of MAFbx/atrogin-1 and MuRF1 through membrane-initiated cortisol action in the skeletal muscle of rainbow trout.
Glucocorticoids are key stress-related hormones in vertebrates, with cortisol being the main glucocorticoid in teleosts. Glucocorticoids exert their effects through two mechanisms of action: genomic/classic and membrane initiated. In mammals, cortisol-mediated stress has been found to be associated with increased expression of critical atrophy-related genes (atrogenes), such as MAFbx/atrogin-1 and murf1/trim63. However, the direct impact of cortisol on the early regulation of atrogene expression in teleost skeletal muscle and the contribution of membrane-initiated cortisol action to this process have not been identified. In this work, the mRNA levels of atrogin-1 and murf1 were assessed in isolated myotubes and skeletal muscle of rainbow trout administered with cortisol or cortisol-BSA. This latter compound is a membrane-impermeable cortisol analog that exclusively induces membrane-initiated effects. We found that cortisol (10 mg/kg) first decreased the expression of both atrogenes at 3 h of treatment and then increased their expression at 9 h of treatment in the skeletal muscle of rainbow trout. Additionally, the in vitro analysis suggested that membrane-initiated cortisol action regulates murf1 but not atrogin-1 in rainbow trout myotubes. Using RU486 to selectively block glucocorticoid receptor (GR), we found that early downregulation of murf1 is potentially mediated by membrane GR signaling in myotubes. Considering the results of both the in vivo and in vitro approaches, we suggest that membrane-initiated cortisol action regulates the early expression of atrophy-related processes in teleosts.


Hormone replacement therapy after endometrial cancer.
Endometrial cancer is the fourth most common female cancer in the UK and the most common gynaecological cancer. Quality of life and symptom control needs to be considered in women who enter a surgically induced menopause. Hormone replacement in this population has been controversial to date. The current evidence regarding the safety of estrogen only and combined hormone replacement therapy is discussed in this review. The use of topical vaginal therapies, alternate therapies and the current data regarding testosterone use for symptom control is also outlined.


Colon tumor cell growth-inhibitory activity of sulindac sulfide and other nonsteroidal anti-inflammatory drugs is associated with phosphodiesterase 5 inhibition.
Nonsteroidal anti-inflammatory drugs (NSAID) display promising antineoplastic activity, but toxicity resulting from cyclooxygenase (COX) inhibition limits their clinical use for chemoprevention. Studies suggest that the mechanism may be COX independent, although alternative targets have not been well defined. Here, we show that the NSAID sulindac sulfide (SS) inhibits cyclic guanosine 3',5'-monophosphate (cGMP) phosphodiesterase (PDE) activity in colon tumor cell lysates at concentrations that inhibit colon tumor cell growth in vitro and in vivo. A series of chemically diverse NSAIDs also inhibited cGMP hydrolysis at concentrations that correlate with their potency to inhibit colon tumor cell growth, whereas no correlation was observed with COX-2 inhibition. Consistent with its selectivity for inhibiting cGMP hydrolysis compared with cyclic AMP hydrolysis, SS inhibited the cGMP-specific PDE5 isozyme and increased cGMP levels in colon tumor cells. Of numerous PDE isozyme-specific inhibitors evaluated, only the PDE5-selective inhibitor MY5445 inhibited colon tumor cell growth. The effects of SS and MY5445 on cell growth were associated with inhibition of β-catenin-mediated transcriptional activity to suppress the synthesis of cyclin D and survivin, which regulate tumor cell proliferation and apoptosis, respectively. SS had minimal effects on cGMP PDE activity in normal colonocytes, which displayed reduced sensitivity to SS and did not express PDE5. PDE5 was found to be overexpressed in colon tumor cell lines as well as in colon adenomas and adenocarcinomas compared with normal colonic mucosa. These results suggest that PDE5 inhibition, cGMP elevation, and inhibition of β-catenin transcriptional activity may contribute to the chemopreventive properties of certain NSAIDs.


Electron microscope observations on synaptic vesicles in synapses of the retinal rods and cones.
The submicroscopic organization of the rod and cone synapses of the albino rabbit has been investigated with the use of the electron microscope. The most common rod synapse consists of an enlarged expansion of the rod fiber (the so called spherule) into which the dendritic postsynaptic fiber of the bipolar cell penetrates and digitates. The membrane surrounding the terminal consists of a double layer, the external of which is interpreted as belonging to the intervening glial cells. The synaptic membrane has a pre- and a postsynaptic layer with a total thickness of 180 to 300 A. The presynaptic layer is frequently denser and is intimately associated with the adjacent synaptic vesicles. The synaptic membrane shows processes constituted by foldings of the presynaptic layer. The entire spherule is filled with synaptic vesicles varying in diameter between 200 and 650 A with a mean of 386 A. In addition, the spherule contains a few large vacuoles near the rod fiber, interpreted as endoplasmic reticulum, and a matrix in which with high resolution a fine filamentous material can be observed. The postsynaptic fiber is homogeneous and usually does not show synaptic vesicles. In animals maintained in complete darkness for 24 hours vesicles appear to accumulate near the synaptic membrane and its processes. After 9 days there is a sharp decrease in size of the synaptic vesicles. A special rod synapse in which the dendritic postsynaptic expansion penetrates directly into the rod cell body has been identified. In line with Cajal's classification this type of synapse could be considered as a somatodendritic one. The cone synapse has a much larger terminal with a more complex relationship with the postsynaptic fiber. However, the same components recognized in the rod synapse can be observed. In animals maintained for 9 days in complete darkness there is also a considerable diminution in size of the synaptic vesicles.


Cross-sectional associations of sedentary behaviour and physical activity on depression in Japanese older adults: an isotemporal substitution approach.
Reducing sedentary behaviour (SB) and increasing physical activity (PA) have been shown to be associated with decreased depression. However, there are yet few studies examining the potential benefits on older adults' depression, when SB is replaced with PA. This study aimed to examine the associations of objectively assessed SB, light-intensity PA (LPA) and moderate-to-vigorous PA (MVPA) with depression among a sample of Japanese older adults, and to explore impacts of substituting SB with PA on older adults' depression. Cross-sectional analysis. General community. A total of 276 older adults aged 65-85 years living in Japan. Three behaviours including the average daily time spent in SB (≤1.5 METs); LPA (>1.5 to <3.0 METs) and MVPA (≥3.0 METs) per day were calculated by accelerometers. Depression was assessed using the Japanese version of the 15-item Geriatric Depression Scale (GDS-15). Less SB (β=0.129, 95% CI 0.015 to 0.243) and more LPA (β=-0.138, 95% CI -0.265 to -0.011) were found to be significantly and negatively associated with the GDS-15 score in the single-activity model. The isotemporal substitution model found that replacing only 30 min per day of SB with the same amount of LPA to be significantly and negatively associated with the GDS-15 score (β=-0.131, 95% CI -0.260 to -0.002). These findings indicated that substituting even small amounts of SB with LPA may contribute to less depression in older adults. Potential favourable effects can be observed for replacing only 30 min per day of SB with LPA.


Lactobacillus rhamnosus cell lysate in the management of resistant childhood atopic eczema.
The number of children suffering from atopic eczema has increased over the past 30 years especially in children between the ages of 2 and 5 years. These is a significant group of eczematous children that are resistant to standard therapy. Babies and children with eczema suffer pain, irritation and disfigurement from the dermatitis. In this study, we have followed 14 cases of pediatric patients (ages of 8 months to 64 months) with a history of resistant eczema for a period of at least six months. All of these children received 300 mg to 500 mg standardized Lactobacillus rhamnosus cell lysate daily as an immunobiotic supplement. The results of this open label non-randomized clinical observation showed a substantial improvement in quality of life, skin symptoms and day- and nighttime irritation scores in children with the supplementation of Lactobacillus rhamnosus lysate. There were no intolerance or adverse reactions observed in these children. Lactobacillus rhamnosus cell lysate may thus be used as a safe and effective immunobiotic for the treatment and prevention of childhood eczema and possible other types of atopy (allergic diseases).


Serum alkaline phosphatase and phosphate and risk of mortality and hospitalization.
Elevated alkaline phosphatase (AlkPhos) and phosphate levels are associated with cardiovascular morbidity and mortality in patients receiving dialysis. A retrospective cohort study was conducted to test these associations in outpatients with an estimated GFR > or =60 ml/min/1.73 m(2). Patients with serum AlkPhos and phosphate levels measured between 2000 and 2002 (n = 10,743) at Montefiore Medical Center (MMC) clinics were followed through September 11, 2008 (median 6.8 years). Mortality data were obtained via Social Security Administration records (n = 949 deaths). Hospitalization data were obtained from MMC records. The mean age was 51 years, 64% were women, 22% were white, 26% were non-Hispanic black, 16% were Hispanic, 13% had a diagnosis of hypertension, 9% had diabetes mellitus, and 8% had cardiovascular disease at baseline. AlkPhos and phosphate were independently associated with mortality and cardiovascular-related hospitalization after multivariable adjustment. Comparing patients in the highest (> or =104 U/L) versus lowest quartile of AlkPhos (< or =66 U/L), the adjusted hazard ratio (HR) for mortality was 1.65 (P trend across quartiles <0.001). For the highest compared with the lowest quartile of serum phosphate (> or =3.8 mg/dl versus < or =3.0 mg/dl), the adjusted HR for mortality was 1.29 (P trend across quartiles = 0.008). High AlkPhos but not phosphate levels were also associated with all-cause, infection-related, and fracture-related hospitalization. Higher levels of serum AlkPhos and phosphate were associated with increased mortality and cardiovascular-related hospitalization in an inner-city clinic population. Further studies are needed to elucidate mechanisms underlying these associations.


A Survey of Chronic Pain Due to Spinal Dural Arteriovenous Fistulae.
Chronic pain due to spinal dural arteriovenous fistulae (SDAVF) during follow-up is a serious issue because it can affect patients' quality of life. The severity of posttreatment chronic leg pain in patients with SDAVF is unclear. To investigate the prevalence, severity, and characteristics of chronic leg pain in patients with SDAVF and to examine the clinical factors associated with chronic pain. We conducted a survey of their chronic posttreatment leg pain in 49 SDAVF patients. It consisted of a numeric rating scale and a neuropathic pain symptom inventory (NPSI). Of the 49 patients, 32 (65.3%) completed the questionnaire. There were 24 males and 8 females whose mean age was 69.1 years. The mean follow-up period was 86.3 months. Chronic leg pain was reported by 26 (81.3%) of 32 patients; 14 experienced pain before treatment and 12 had new-onset posttreatment pain. The pain was reported as moderate to severe by 21 (80.8%) of 26 patients; the mean numeric rating scale and NPSI scores were 6.0 ± 2.3 and 17.4 ± 10.3, respectively. The NPSI subscores were significantly higher for spontaneous pain and paresthesia/dysesthesia than for paroxysmal pain. We found no significant clinical factors related to chronic posttreatment pain; on magnetic resonance imaging scans we identified spinal cord atrophy in 8 patients with chronic pain. Most of our SDAVF patients reported moderate to severe chronic leg pain characterized by spontaneous pain and paresthesia/dysesthesia. Spinal cord atrophy on magnetic resonance imaging scans was a characteristic in patients with chronic pain.


Mortality patterns in infectious salmon anaemia virus outbreaks in New Brunswick, Canada.
Mortality levels attributed to infectious salmon anaemia viral (ISAV) infections were examined at the net pen and site level in the 1996 smolt year class in three areas of New Brunswick, Canada. The year class in this region was the first known to have potential exposure to ISAV beginning at the time of seawater transfer. There was considerable variability in mortality patterns among net pen groups of fish. Net pen outbreak definitions were based on at least seven high mortality days in which there were at least 100 per 100 000 fish per day or >5% cumulative mortality for the study period. There were 106 net pen outbreaks in a study population consisting of 218 net pens. Although the number of new cases decreased as water temperature decreased, overall mortality levels at the study sites did not decrease noticeably. The median peak daily mortality rate during outbreaks was 492 per 100 000 fish per day, with 10% of cases experiencing >5200 mortalities per 100 000 fish per day. The median duration of outbreaks in net pens for which the fish were not slaughtered during the outbreak was 33 days and the median total loss in those outbreaks was 6600 per 100 000 fish.


Plasmodium falciparum dihydrofolate reductase and dihyropteroate synthase mutations and the use of trimethoprim-sulfamethoxazole prophylaxis among persons infected with human immunodeficiency virus.
A prospective cohort design was used to measure the association between daily cotrimoxazole-prophylaxis and infection with Plasmodium falciparum containing mutations associated with antifolate resistance among persons infected with human immunodeficiency virus (HIV) in Tororo and Busia District, in eastern Uganda. Of 149 cases of P. falciparum parasitemia diagnosed, 147 (99%) (smears from participants taking prophylaxis = 91 and smears from those not taking cotrimoxazole prophylaxis = 56) were successfully assessed for mutations in the dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) mutations associated with antifolate resistance. Prevalences of the dhfr pure triple mutant (74% and 70%; P = 0.71), the dhps pure double mutant (95% and 88%; P = 0.21), and the dhfr/dhps pure quintuple mutant (73% and 64%; P = 0.36), were not significantly different between those taking and those not taking cotrimoxazole-prophylaxis, respectively. The overall prevalence of the pure quintuple mutant in this study was 69%, which is among the highest in Africa. Although resistance rates of P. falciparum to antifolate drugs are high, cotrimoxazole-prophylaxis in HIV-infected persons was not associated with a higher prevalence of mutations associated with antifolate resistance.


A randomized controlled trial of antidepressant continuation for major depression following traumatic brain injury.
This study examines whether continuation therapy with citalopram can prevent a relapse following remission of major depression due to traumatic brain injury. After 65 subjects with DSM-IV-diagnosed major depression following traumatic brain injury were treated with open-label citalopram (20 mg to 50 mg/d), 25 subjects (38.5%) met criteria for remission. Of those, 21 (84.0%) were randomly assigned to either same-dose citalopram or placebo and followed monthly over 40 weeks. Remission was defined as a Hamilton Depression Rating Scale (HDRS) score of ≤ 7 or a Clinical Global Impressions-Improvement rating of "much improved" or better. The main outcome variable was the presence of relapse, as defined by meeting criteria for major depressive episode according to the DSM-IV and an HDRS score ≥ 16. Data were collected from February 16, 2005, to May 5, 2008. Ten subjects were randomly assigned to citalopram and 11 to placebo. There were 3 dropouts, including 1 for adverse drug effects (diarrhea). Relapse occurred in 11 subjects (52.4%), with a mean ± SD time to relapse of 23.52 ± 16.6 weeks. The groups did not differ in relapse rates (drug: 50.0% [5/10] vs placebo: 54.5% [6/11], Fisher exact test, P = .835) or time to relapse (log rank test χ² = 0.148, P = .700). The present study suggests important limitations of continuation pharmacotherapy in the prevention of relapse of major depression following traumatic brain injury. clinicaltrials.gov Identifier: NCT00162916.


BACE inhibition causes rapid, regional, and non-progressive volume reduction in Alzheimer's disease brain.
In the phase 3 EPOCH trial (Clinicaltrials.gov; NCT01739348), treatment with the BACE inhibitor verubecestat failed to improve cognition in patients with mild-to-moderate Alzheimer's disease, but was associated with reduced hippocampal volume after 78 weeks as assessed by MRI. The aims of the present exploratory analyses were to: (i) characterize the effect of verubecestat on brain volume by evaluating the time course of volumetric MRI changes for a variety of brain regions; and (ii) understand the mechanism through which verubecestat might cause hippocampal (and other brain region) volume loss by assessing its relationship to measures of amyloid, neurodegeneration, and cognition. Participants were aged 55-85 years with probable Alzheimer's disease dementia and a Mini Mental State Examination score ≥15 and ≤26. MRIs were obtained at baseline and at Weeks 13, 26, 52 and 78 of treatment. MRIs were segmented using Freesurfer and analysed using a tensor-based morphometry method. PET amyloid data were obtained with 18F-flutemetamol (Vizamyl®) at baseline and Week 78. Standardized uptake value ratios were generated with subcortical white matter as a reference region. Neurofilament light chain in the CSF was assessed as a biomarker of neurodegeneration. Compared with placebo, verubecestat showed increased MRI brain volume loss at Week 13 with no evidence of additional loss through Week 78. The verubecestat-related volumetric MRI loss occurred predominantly in amyloid-rich brain regions. Correlations between amyloid burden at baseline and verubecestat-related volumetric MRI reductions were not significant (r = 0.05 to 0.26, P-values > 0.27). There were no significant differences between verubecestat and placebo in changes from baseline in CSF levels of neurofilament light chain at Week 78 (increases of 7.2 and 14.6 pg/ml for verubecestat versus 19.7 pg/ml for placebo, P-values ≥ 0.1). There was a moderate correlation between volumetric MRI changes and cognitive decline in all groups including placebo at Week 78 (e.g. r = -0.45 to -0.55, P < 0.001 for whole brain), but the correlations were smaller at Week 13 and significant only for the verubecestat groups (e.g. r = -0.15 and -0.11, P < 0.04 for whole brain). Our results suggest that the verubecestat-associated MRI brain volume loss is not due to generalized, progressive neurodegeneration, but may be mediated by specific effects on BACE-related amyloid processes.


Shifted T Helper Cell Polarization in a Murine Staphylococcus aureus Mastitis Model.
Mastitis, one of the most costly diseases in dairy ruminants, is an inflammation of the mammary gland caused by pathogenic infection. The mechanisms of adaptive immunity against pathogens in mastitis have not been fully elucidated. To investigate T helper cell-mediated adaptive immune responses, we established a mastitis model by challenge with an inoculum of 4 × 106 colony-forming units of Staphylococcus aureus in the mammary gland of lactating mice, followed by quantification of bacterial burden and histological analysis. The development of mastitis was accompanied by a significant increase in both Th17 and Th1 cells in the mammary gland. Moreover, the relative expression of genes encoding cytokines and transcription factors involved in the differentiation and function of these T helper cells, including Il17, Rorc, Tgfb, Il1b, Il23, Ifng, Tbx21, and Il12, was greatly elevated in the infected mammary gland. IL-17 is essential for neutrophil recruitment to infected mammary gland via CXC chemokines, whereas the excessive IL-17 production contributes to tissue damage in mastitis. In addition, a shift in T helper cell polarization toward Th2 and Treg cells was observed 5 days post-infection, and the mRNA expression of the anti-inflammatory cytokine Il10 was markedly increased at day 7 post-infection. These results indicate that immune clearance of Staphylococcus aureus in mastitis is facilitated by the enrichment of Th17, Th1 and Th2 cells in the mammary gland mediated by pro-inflammatory cytokine production, which is tightly regulated by Treg cells and the anti-inflammatory cytokine IL-10.


Self-injurious behavior in young boys with fragile X syndrome.
In this study, we distributed surveys to 67 families of young boys with fragile X syndrome to determine the prevalence, onset, form, function, location, and correlates of self-injurious behavior. Fifty-five surveys were completed (82%). The mean age of the boys at the time of the survey was 80 months (range = 20-144). Self-injurious behavior (SIB) was reported for 58% of the participants with a mean age of onset of 31 months. The mean number of forms of self-injury was 2 per participant. Biting was the most commonly reported form of self-injury with the fingers and back of the hand disproportionately targeted as the most prevalent self-injury body site. There was no linear increase in risk of SIB with age past 25 months. SIB was reported as most likely to occur following the presentation of difficult task demands or changes in routine. Significant group differences were found between overall ratings of problem behavior for boys with self-injury compared to those without self-injury. Groups did not differ on measures of fragile X mental retardation protein (FMRP), autism status, adaptive behavior, or age first medicated. Results are discussed in terms of future research designed to further elucidate the behavioral phenotype of fragile X syndrome.


Counsellors' perspectives on the experience of providing HIV counselling in Kenya and Tanzania: the Voluntary HIV-1 Counselling and Testing Efficacy Study.
Demand for HIV counselling services is increasing in developing counties, but there have been few previous studies that describe counsellors' roles and experiences providing HIV-related counselling in developing countries. Such information can be used to better supervise and support counsellors and thereby improve counselling services. As a sub-study of the Voluntary Counseling and Testing Efficacy Study, we conducted focus groups and individual interviews with 11 counsellors and counselling supervisors providing HIV counselling services in Kenya and Tanzania. Counsellors told us that their jobs were both rewarding and stressful. In addition to their obligations in the counselling relationship (providing information, protecting confidentiality and being non-judgemental), they perceived pressure to provide information and be good role models in their communities. Additional stresses were related to external (economic and political) conditions, 'spillover' of HIV issues from their personal lives and providing counselling in a research setting. Counsellor stress might be reduced and their effectiveness and retention improved by (1) allowing work flexibility; (2) providing supportive, non-evaluative supervision; (3) offering alternatives to client behaviour change as the indication of counsellor performance; (4) acknowledging and educating about 'emotional labour' in counselling; (5) providing frequent information updates and intensive training; and (6) encouraging counsellor participation in the development of research protocols.


Association of education with dietary intake among young adults in the bi-ethnic Coronary Artery Risk Development in Young Adults (CARDIA) cohort.
To examine associations of changes in dietary intake with education in young black and white men and women. The Coronary Artery Risk Development in Young Adults (CARDIA) study, a multi-centre population-based prospective study. Dietary intake data at baseline and year 7 were obtained from an extensive nutritionist-administered diet history questionnaire with 700 items developed for CARDIA. Participants were recruited in 1985-1986 from four sites: Birmingham, Alabama; Chicago, Illinois; Minneapolis, Minnesota; and Oakland, California. Participants were from a general community sample of 703 black men (BM), 1006 black women (BW), 963 white men (WM) and 1054 white women (WW) who were aged 18-30 years at baseline. Analyses here include data for baseline (1985-1986) and year 7 (1992-1993). Most changes in dietary intake were observed among those with high education (>or=12 years) at both examinations. There was a significant decrease in intake of energy from saturated fat and cholesterol and a significant increase in energy from starch for each race-gender group (P<0.001). Regardless of education, taste was considered an important influence on food choice. The inverse relationship of education with changes in saturated fat and cholesterol intakes suggests that national public health campaigns may have a greater impact among those with more education.


[Caries prevalence among students in the city of Winterthur: comparison with the city of Zurich and 16 communities in the canton of Zurich].
The aim of the present survey was to assess the caries prevalence in schoolchildren of the city of Winterthur in the year 2002. A simple random sample of 1000 schoolchildren was drawn. 65% of the children participated in the standardized investigation. The mean DMFT-value (permanent dentition) in 12-year-olds was 1.0 and was on a low level in a European comparison. The mean dmft-value (deciduous dentition) in 7-year-olds was 2.0. From 1971 to 2002, DMFT-values decreased by about 80% in Winterthur. The dmft-values decreased by about 60%. The splitting by nationality revealed a high caries prevalence in schoolchildren from the former Yugoslavia and Albania. A multivariate analysis showed no significant differences in caries prevalence between the schoolchildren of the city of Winterthur and other communities of the canton of Zurich.


Treatment utilization by patients with personality disorders.
Utilization of mental health treatment was compared in patients with personality disorders and patients with major depressive disorder without personality disorder. Semistructured interviews were used to assess diagnosis and treatment history of 664 patients in four representative personality disorder groups-schizotypal, borderline, avoidant, and obsessive-compulsive-and in a comparison group of patients with major depressive disorder. Patients with personality disorders had more extensive histories of psychiatric outpatient, inpatient, and psychopharmacologic treatment than patients with major depressive disorder. Compared to the depression group, patients with borderline personality disorder were significantly more likely to have received every type of psychosocial treatment except self-help groups, and patients with obsessive-compulsive personality disorder reported greater utilization of individual psychotherapy. Patients with borderline personality disorder were also more likely to have used antianxiety, antidepressant, and mood stabilizer medications, and those with borderline or schizotypal personality disorder had a greater likelihood of having received antipsychotic medications. Patients with borderline personality disorder had received greater amounts of treatment, except for family/couples therapy and self-help, than the depressed patients and patients with other personality disorders. These results underscore the importance of considering personality disorders in diagnosis and treatment of psychiatric patients. Borderline and schizotypal personality disorder are associated with extensive use of mental health resources, and other, less severe personality disorders may not be addressed sufficiently in treatment planning. More work is needed to determine whether patients with personality disorders are receiving adequate and appropriate mental health treatments.


Oral bisphosphonate use and breast cancer incidence in postmenopausal women.
Emerging clinical evidence suggests intravenous bisphosphonates may inhibit breast cancer while oral bisphosphonates have received limited evaluation regarding breast cancer influence. The association between oral bisphosphonate use and invasive breast cancer was examined in postmenopausal women enrolled onto the Women's Health Initiative (WHI). We compared a published hip fracture prediction model, which did not incorporate bone mineral density (BMD), with total hip BMD in 10,418 WHI participants who had both determinations. To adjust for potential BMD difference based on bisphosphonate use, the hip fracture prediction score was included in multivariant analyses as a BMD surrogate. Of the 154,768 participants, 2,816 were oral bisphosphonate users at entry (90% alendronate, 10% etidronate). As calculated hip fracture risk score was significantly associated with both BMD (regression line = 0.79 to 0.0478 log predicted fracture; P < .001; r = 0.43) and breast cancer incidence (P = .03), this variable was incorporated into regression analyses to adjust for BMD difference between users and nonusers of bisphopshonate. After 7.8 mean years of follow-up (standard deviation, 1.7), invasive breast cancer incidence was lower in bisphosphonate users (hazard ratio [HR], 0.68; 95% CI, 0.52 to 0.88; P < .01) as was incidence of estrogen receptor (ER) -positive invasive cancers (HR, 0.70; 95% CI, 0.52 to 0.94, P = .02). A similar but not significant trend was seen for ER-negative invasive cancers. The incidence of ductal carcinoma in situ was higher in bisphosphonate users (HR, 1.58; 95% CI, 1.08 to 2.31; P = .02). Oral bisphosphonate use was associated with significantly lower invasive breast cancer incidence, suggesting bisphosphonates may have inhibiting effects on breast cancer.


A novel model system for design of biomaterials based on recombinant analogs of spider silk proteins.
Spider dragline silk possesses impressive mechanical and biochemical properties. It is synthesized by a couple of major ampullate glands in spiders and comprises of two major structural proteins--spidroins 1 and 2. The relationship between structure and mechanical properties of spider silk is not well understood. Here, we modeled the complete process of the spider silk assembly using two new recombinant analogs of spidroins 1 and 2. The artificial genes sequence of the hydrophobic core regions of spidroin 1 and 2 have been designed using computer analysis of existing databases and mathematical modeling. Both proteins were expressed in Pichia pastoris and purified using a cation exchange chromatography. Despite the absence of hydrophilic N- and C-termini, both purified proteins spontaneously formed the nanofibrils and round micelles of about 1 microm in aqueous solutions. The electron microscopy study has revealed the helical structure of a nanofibril with a repeating motif of 40 nm. Using the electrospinning, the thin films with an antiparallel beta-sheet structure were produced. In summary, we were able to obtain artificial structures with characteristics that are perspective for further biomedical applications, such as producing three-dimensional matrices for tissue engineering and drug delivery.


Intravenously administered oligonucleotides can be delivered to conducting airway epithelium via the bronchial circulation.
Topical gene transfer to the airways of cystic fibrosis (CF) patients has been inefficient, partly due to extracellular barriers such as sputum. In an attempt to circumvent these, we assessed whether airway epithelial cells can be transfected by intravenous (i.v.) administration of liposome-complexed or "naked" oligonucleotides (ODNs). The conducting airways are the likely target for CF therapy and are supplied by the bronchial circulation. Consequently, we assessed ODN transfer in the mouse trachea and main bronchi as these are supplied by the bronchial circulation. Liposome-protamine-DNA (LPD) complexes were detected in the bronchial circulation but did not transfect conducting airway epithelial cells, even in the presence of microvascular leakage. In contrast, 'naked' ODNs were delivered to 17% (inter-quartile range (IQR) 10-34%) and 35% (IQR 24-59%) of epithelial cells when injected at 500 microg/animal, without and with microvascular leakage, respectively. Two types of nuclear signal were observed; punctate in cells throughout the airways (3%, IQR 2-6%, and 6%, IQR 4-7%, of cells when delivered without and with microvascular leakage, respectively) and diffuse in a small number of epithelial cells in the proximal trachea. ODNs may be relevant to CF in a variety of ways and these data suggest one way towards implementing their use.


Association between high cost user status and end-of-life care in hospitalized patients: A national cohort study of patients who die in hospital.
Studies comparing end-of-life care between patients who are high cost users of the healthcare system compared to those who are not are lacking. The objective of this study was to describe and measure the association between high cost user status and several health services outcomes for all adults in Canada who died in acute care, compared to non-high cost users and those without prior healthcare use. We used administrative data for all adults who died in hospital in Canada between 2011 and 2015 to measure the odds of admission to the intensive care unit (ICU), receipt of invasive interventions, major surgery, and receipt of palliative care during the hospitalization in which the patient died. High cost users were defined as those in the top 10% of acute healthcare costs in the year prior to a person's hospitalization in which they died. Among 252,648 people who died in hospital, 25,264 were high cost users (10%), 112,506 were non-high cost users (44.5%) and 114,878 had no prior acute care use (45.5%). After adjustment for age and sex, high cost user status was associated with a 14% increased odds of receiving an invasive intervention, a 15% increased odds of having major surgery, and an 8% lower odds of receiving palliative care compared to non-high cost users, but opposite when compared to patients without prior healthcare use. Many patients receive aggressive elements of end-of-life care during the hospitalization in which they die and a substantial number do not receive palliative care. Understanding how this care differs between those who were previously high- and non-high cost users may provide an opportunity to improve end of life care for whom better care planning and provision ought to be an equal priority.


Damages with High Consequences: Analysis of Perforations in Surgical Latex Operation Gloves from Orthopedic Surgeries.
To prevent surgical site infections (SSIs) during operation, the use of sterile surgical latex gloves is common. The aim of this study was to examine the damage of the gloves in surgeries with different mechanical stress and the influence on the kind of damages. Gloves were collected during primary arthroplasty, revision arthroplasty (hip and knee), and arthroscopy (shoulder, hip, and knee). Surgical latex operation gloves were collected from surgeons after the operation and were tested with watertightness test (ISO EN 455-1:2000). A total of 1460 surgical gloves were retrieved from 305 elective operations. On average, 15.9% of the gloves showed postoperative lesions, with the highest incidence occurring in revision arthroplasty with 25%. In primary and revision arthroplasty, the index finger of the dominant hand was most frequently affected (62.7% and 58.6%); in contrast, gloves from arthroscopies had most lesions on thumb and middle finger (42.9% each). Tear and perforation size differed from ≤1 mm to >5 mm, and primary and revision arthroplasty showed bigger damages. Surgical gloves have a high malfunction, which increases with growing mechanical stress. A high rate of perforation occurred mostly in revision arthroplasty. Breaching the integrity of the gloves, especially by high mechanical loads, could lead to an increased rate of infection.


Estimating damage costs of flooding on small- and medium-sized enterprises in Kigali, Rwanda.
In the past, Kigali has frequently experienced heavy rain events. These have often led to flooding, which also affected businesses. In the face of climate change, such events can become more frequent and can threaten economic development. To determine if more action is needed to protect businesses from flooding, we assessed how many businesses have suffered damages from floods in certain years in a certain area of Kigali. We also gathered information on how businesses were affected, how they are preparing for flooding and what support they are seeking. We developed and piloted a survey, a standardised questionnaire for gathering information on the relevance of flooding for businesses. The survey was then conducted among 350 businesses in Kigali asking business owners about their experiences with flooding in recent years. Eighty-one per cent of businesses have been affected by floods in 2013 and 2014. The annual damage costs resemble 22% of the total net profit of the businesses in the area. The most common damages were damages to goods that were to be sold and damages to buildings. The extent of past flood damages warrants action on flood risk management, both by businesses and citizens, as well as by city officials. Suitable actions range from increasing awareness about suitable protection measures to upgrading the sewage system.


Comparisons of computational methods for differential alternative splicing detection using RNA-seq in plant systems.
Alternative Splicing (AS) as a post-transcription regulation mechanism is an important application of RNA-seq studies in eukaryotes. A number of software and computational methods have been developed for detecting AS. Most of the methods, however, are designed and tested on animal data, such as human and mouse. Plants genes differ from those of animals in many ways, e.g., the average intron size and preferred AS types. These differences may require different computational approaches and raise questions about their effectiveness on plant data. The goal of this paper is to benchmark existing computational differential splicing (or transcription) detection methods so that biologists can choose the most suitable tools to accomplish their goals. This study compares the eight popular public available software packages for differential splicing analysis using both simulated and real Arabidopsis thaliana RNA-seq data. All software are freely available. The study examines the effect of varying AS ratio, read depth, dispersion pattern, AS types, sample sizes and the influence of annotation. Using a real data, the study looks at the consistences between the packages and verifies a subset of the detected AS events using PCR studies. No single method performs the best in all situations. The accuracy of annotation has a major impact on which method should be chosen for AS analysis. DEXSeq performs well in the simulated data when the AS signal is relative strong and annotation is accurate. Cufflinks achieve a better tradeoff between precision and recall and turns out to be the best one when incomplete annotation is provided. Some methods perform inconsistently for different AS types. Complex AS events that combine several simple AS events impose problems for most methods, especially for MATS. MATS stands out in the analysis of real RNA-seq data when all the AS events being evaluated are simple AS events.


Production, purification and characterization of recombinant, full-length human claudin-1.
The transmembrane domain proteins of the claudin superfamily are the major structural components of cellular tight junctions. One family member, claudin-1, also associates with tetraspanin CD81 as part of a receptor complex that is essential for hepatitis C virus (HCV) infection of the liver. To understand the molecular basis of claudin-1/CD81 association we previously produced and purified milligram quantities of functional, full-length CD81, which binds a soluble form of HCV E2 glycoprotein (sE2). Here we report the production, purification and characterization of claudin-1. Both yeast membrane-bound and detergent-extracted, purified claudin-1 were antigenic and recognized by specific antibodies. Analytical ultracentrifugation demonstrated that extraction with n-octyl-β-d-glucopyranoside yielded monodispersed, dimeric pools of claudin-1 while extraction with profoldin-8 or n-decylphosphocholine yielded a dynamic mixture of claudin-1 oligomers. Neither form bound sE2 in line with literature expectations, while further functional analysis was hampered by the finding that incorporation of claudin-1 into proteoliposomes rendered them intractable to study. Dynamic light scattering demonstrated that claudin-1 oligomers associate with CD81 in vitro in a defined molar ratio of 1∶2 and that complex formation was enhanced by the presence of cholesteryl hemisuccinate. Attempts to assay the complex biologically were limited by our finding that claudin-1 affects the properties of proteoliposomes. We conclude that recombinant, correctly-folded, full-length claudin-1 can be produced in yeast membranes, that it can be extracted in different oligomeric forms that do not bind sE2 and that a dynamic preparation can form a specific complex with CD81 in vitro in the absence of any other cellular components. These findings pave the way for the structural characterization of claudin-1 alone and in complex with CD81.


Scope, quality and inclusivity of international clinical guidelines on mental health and substance abuse in relation to dual diagnosis, social and community outcomes: a systematic review.
It is estimated that up to 75% of patients with severe mental illness (SMI) also have substance use disorder (SUD). The aim of this systematic review was to explore the scope, quality and inclusivity of international clinical guidelines on mental health and/or substance abuse in relation to diagnosis and treatment of co-existing disorders and considerations for wider social and contextual factors in treatment recommendations. A protocol (PROSPERO CRD42020187094) driven systematic review was conducted. A systematic search was undertaken using six databases including MEDLINE, Cochrane Library, EMBASE, PsychInfo from 2010 till June 2020; and webpages of guideline bodies and professional societies. Guideline quality was assessed based on 'Appraisal of Guidelines for Research & Evaluation II' (AGREE II) tool. Data was extracted using a pre-piloted structured data extraction form and synthesized narratively. Reporting was based on PRISMA guideline. A total of 12,644 records were identified. Of these, 21 guidelines were included in this review. Three of the included guidelines were related to coexisting disorders, 11 related to SMI, and 7 guidelines were related to SUD. Seven (out of 18) single disorder guidelines did not adequately recommend the importance of diagnosis or treatment of concurrent disorders despite their high co-prevalence. The majority of the guidelines (n = 15) lacked recommendations for medicines optimisation in accordance with concurrent disorders (SMI or SUD) such as in the context of drug interactions. Social cause and consequence of dual diagnosis such as homelessness and safeguarding and associated referral pathways were sparsely mentioned. Despite very high co-prevalence, clinical guidelines for SUD or SMI tend to have limited considerations for coexisting disorders in diagnosis, treatment and management. There is a need to improve the scope, quality and inclusivity of guidelines to offer person-centred and integrated care.


Semi-parametric proportional hazards models with crossed random effects for psychometric response times.
The semi-parametric proportional hazards model with crossed random effects has two important characteristics: it avoids explicit specification of the response time distribution by using semi-parametric models, and it captures heterogeneity that is due to subjects and items. The proposed model has a proportionality parameter for the speed of each test taker, for the time intensity of each item, and for subject or item characteristics of interest. It is shown how all these parameters can be estimated by Markov chain Monte Carlo methods (Gibbs sampling). The performance of the estimation procedure is assessed with simulations and the model is further illustrated with the analysis of response times from a visual recognition task.


Frequency domain formulation of active parametric deformable models.
Active deformable models are simple tools, very popular in computer vision and computer graphics, for solving ill-posed problems or mimic real physical systems. The classical formulation is given in the spatial domain, the motor of the procedure is a second-order linear system, and rigidity and elasticity are the basic parameters for its characterization. This paper proposes a novel formulation based on a frequency-domain analysis: The internal energy functional and the Lagrange minimization are performed entirely in the frequency domain, which leads to a simple formulation and design. The frequency-based implementation offers important computational savings in comparison to the original one, a feature that is improved by the efficient hardware and software computation of the FFT algorithm. This new formulation focuses on the stiffness spectrum, allowing the possibility of constructing deformable models apart from the elasticity and rigidity-based original formulation. Simulation examples validate the theoretical results.


StressHacker: Towards Practical Stress Monitoring in the Wild with Smartwatches.
In modern life, the nonstop and pervasive stress tends to keep us on long-lasting high alert, which over time, could lead to a broad range of health problems from depression, metabolic disorders to heart diseases. However, there is a stunning lack of practical tools for effective stress management that can help people navigate through their daily stress. This paper presents the feasibility evaluation of StressHacker, a smartwatch-based system designed to continuously and passively monitor one's stress level using bio-signals obtained from the on-board sensors. With the proliferation of smartwatches, StressHacker is highly accessible and suited for daily use. Our preliminary evaluation is based on 300 hours of data collected in a real-life setting (12 subjects, 29 days). The result suggests that StressHacker is capable of reliably capturing daily stress dynamics (precision = 86.1%, recall = 91.2%), thus with great potential to enable seamless and personalized stress management.


Farnesyl protein transferase inhibitors as potential cancer chemopreventives.
Among the most important targets for chemopreventive intervention and drug development are deregulated signal transduction pathways. Ras proteins serve as central connectors between signals generated at the plasma membrane and nuclear effectors; thus, disrupting the Ras signaling pathway could have significant potential as a cancer chemopreventive strategy. Target organs for Ras-based chemopreventive strategies include those associated with activating ras mutations (e.g., colorectum, pancreas, and lung) and those carrying aberrations in upstream element(s), such as growth factors and their receptors. Ras proteins require posttranslational modification with a farnesyl moiety for both normal and oncogenic activity. Inhibitors of the enzyme that catalyzes this reaction, farnesyl protein transferase (FPT) should, therefore, inhibit Ras-dependent proliferative activity in cancerous and precancerous lesions (J. B. Gibbs et al., Cell, 77: 175-178, 1994). Because growth factor networks are redundant, selective inhibition of signaling pathways activated in precancerous and cancerous cells should be possible. Requirements for Ras farnesylation inhibitors include: specificity for FPT compared with other prenyl transferases; specificity for FPT compared with other farnesyl PPi-utilizing enzymes; ability to specifically inhibit processing of mutant K-ras (the most commonly mutated ras gene in human cancers); high potency; selective activity in intact cells; activity in vivo; and lack of toxicity. Numerous FPT inhibitors have been identified through random screening of natural products and by rational design of analogues of the two substrates, farnesyl PPi and the COOH-terminal CAAX motif of Ras tetrapeptides. A possible testing strategy for developing FPT inhibitors as chemopreventive agents includes the following steps: (a) determine FPT inhibitory activity in vitro; (b) evaluate selectivity (relative to other protein prenyl transferases and FPT-utilizing enzymes); (c) determine inhibition of Ras-mediated effects in intact cells; (d) determine inhibition of Ras-mediated effects in vivo (e.g., in nude mouse tumor xenografts); and (e) determine chemopreventive efficacy in vivo (e.g., in carcinogen-induced A/J mouse lung, rat colon, or hamster pancreas).


Modulation of Lactobacillus plantarum gastrointestinal robustness by fermentation conditions enables identification of bacterial robustness markers.
Lactic acid bacteria (LAB) are applied worldwide in the production of a variety of fermented food products. Additionally, specific Lactobacillus species are nowadays recognized for their health-promoting effects on the consumer. To optimally exert such beneficial effects, it is considered of great importance that these probiotic bacteria reach their target sites in the gut alive. In the accompanying manuscript by Bron et al. the probiotic model organism Lactobacillus plantarum WCFS1 was cultured under different fermentation conditions, which was complemented by the determination of the corresponding molecular responses by full-genome transcriptome analyses. Here, the gastrointestinal (GI) survival of the cultures produced was assessed in an in vitro assay. Variations in fermentation conditions led to dramatic differences in GI-tract survival (up to 7-log) and high robustness could be associated with low salt and low pH during the fermentations. Moreover, random forest correlation analyses allowed the identification of specific transcripts associated with robustness. Subsequently, the corresponding genes were targeted by genetic engineering, aiming to enhance robustness, which could be achieved for 3 of the genes that negatively correlated with robustness and where deletion derivatives displayed enhanced survival compared to the parental strain. Specifically, a role in GI-tract survival could be confirmed for the lp_1669-encoded AraC-family transcription regulator, involved in capsular polysaccharide remodeling, the penicillin-binding protein Pbp2A involved in peptidoglycan biosynthesis, and the Na(+)/H(+) antiporter NapA3. Moreover, additional physiological analysis established a role for Pbp2A and NapA3 in bile salt and salt tolerance, respectively. Transcriptome trait matching enabled the identification of biomarkers for bacterial (gut-)robustness, which is important for our molecular understanding of GI-tract survival and could facilitate the design of culture conditions aimed to enhance probiotic culture robustness.


Sulfonylureas rapidly cross phospholipid bilayer membranes by a free-diffusion mechanism.
Because sulfonylureas directly activate the exocytotic machinery, we were interested in the extent to which these compounds penetrate the beta-cell plasma membrane and the underlying molecular mechanism(s). We now provide evidence that sulfonylureas cross phospholipid bilayer membranes rapidly and effectively by a free-diffusion mechanism. Two sulfonylurea compounds investigated by 1H nuclear magnetic resonance spectroscopy, glibenclamide and tolbutamide, were found to incorporate into phospholipid bilayers, with the ionizable sulfonamide exposed to the aqueous interface and its apparent dissociation constant (pKa) increased to approximately 7.0. Diffusion of weak amphiphilic acids across membranes is associated with a measurable change in pH. Thus, by using a fluorescence-based pH assay, we could investigate the diffusion of sulfonylurea compounds across phospholipid bilayer membranes. A fluorescent pH indicator (pyranin or [2',7'-bis (2-carboxyethyl)-5(6)-carboxyfluorescein] [BCECF]) was trapped in egg phosphatidylcholine vesicles. Addition of glibenclamide decreased internal pH (pHin), and addition of albumin reversed this drop by 50%. With the same amount of tolbutamide, the decrease in pHin was much smaller, primarily because of the lower partitioning of tolbutamide into phospholipid bilayers. Using similar protocols, we also demonstrated diffusion by the same mechanism across the beta-cell plasma membrane. Thus, we now provide a molecular mechanism by which sulfonylureas can penetrate the plasma membrane and reach intracellular sites regulating exocytosis.


An Orally Active Bradykinin B1 Receptor Antagonist Engineered as a Bifunctional Chimera of Sunflower Trypsin Inhibitor.
An orally active and metabolically stable peptide TIBA was successfully engineered as a chimera by fusing an analgesic bradykinin receptor antagonist peptide and the trypsin inhibitory loop of sunflower trypsin inhibitor-1. As a fusion cyclic peptide, the metabolically labile analgesic peptide is protected from degradation by exopeptidases as well as the endopeptidases, and its serum half-life extended from <5 min to >6 h as a chimera. Moreover, the chimera TIBA was also found to be orally active in an animal pain model using a hot plate assay.


The physiology of salinity tolerance in larvae of two species of Culex mosquitoes: the role of compatible solutes.
We investigated the physiological basis for differences in salinity tolerance ranges in mosquito larvae of the genus Culex. We examined the response of larvae of C. quinquefasciatus, a freshwater obligate, and C. tarsalis, a euryhaline osmoconformer, following transfer from fresh water to 34% sea water. Hemolymph Na(+) and Cl(-) levels increased similarly in both species, indicating that ion regulation does not differ under these conditions. C. quinquefasciatus responded to increased environmental salinity with increased hemolymph levels of serine, but suffered a significant reduction in levels of trehalose. C. tarsalis responded to increased environmental salinity with increased hemolymph levels of both proline and trehalose. When C. tarsalis larvae were held in 64% sea water, which C. quinquefasciatus larvae cannot tolerate, hemolymph proline and trehalose were accumulated approximately 50-fold and twofold, respectively, relative to freshwater values. We found that proline serves as both an intra- and extracellular compatible solute in C. tarsalis, the first such circumstance documented in an animal in response to increased environmental salinity. Analyses of the acute responses of the two species to an increase in salinity (from 30% to 50% sea water) indicate that larvae of C. tarsalis are able to volume-regulate via drinking and to attenuate increases in hemolymph NaCl concentration using unknown mechanisms during large, rapid increases in salinity.


Closely related yet different: a borylene and its dimer are non-interconvertible but connected through reactivity.
The self-stabilizing, tetrameric cyanoborylene [(cAAC)B(CN)]4 (I, cAAC = 1-(2,6-diisopropylphenyl)-3,3,5,5-tetramethylpyrrolidin-2-ylidene) and its diborene relative, [(cAAC)(CN)B[double bond, length as m-dash]B(CN)(cAAC)] (II), both react with disulfides and diselenides to yield the corresponding cAAC-supported cyanoboron bis(chalcogenides). Furthermore, reactions of I or II with elemental sulfur and selenium in various stoichiometries provided access to a variety of cAAC-stabilized cyanoboron-chalcogen heterocycles, including a unique dithiaborirane, a diboraselenirane, 1,3-dichalcogena-2,4-diboretanes, 1,3,4-trichalcogena-2,5-diborolanes and a rare six-membered 1,2,4,5-tetrathia-3,6-diborinane. Stepwise addition reactions and solution stability studies provided insights into the mechanism of these reactions and the subtle differences in reactivity observed between I and II.


The estrogen metabolites 2-methoxyestradiol and 2-hydroxyestradiol inhibit endometriotic cell proliferation in estrogen-receptor-independent manner.
Endometriosis, a painful disorder associated with infertility, is estimated to occur in approximately 7-10% of reproductive age women. Although endometriosis is considered as an estrogen-dependent disease, the role of estrogen metabolites via receptor-independent mechanisms has not yet been comprehensively clarified. In the present study, growth studies were performed comparing the effect of estradiol (E2), estrogen metabolites, that is, 2-hydroxyestradiol (2-OHE2) and 2-methoxyestradiol (2-ME), as well as estrogen-receptor-independent mechanisms using the estrogen receptor antagonist fulvestrant, on cell proliferation of endometriotic cells. The estrogen metabolites 2-OHE2 and 2-ME inhibited cell growth in a dose-dependent manner in pharmacological doses. Lower concentrations of 2-OHE2 had a stimulating effect on cell proliferation while pharmacologic doses exerted an antimitogenic effect. The effects on cell growth were at least partially receptor-independent, as demonstrated by simultaneous receptor antagonization with fulvestrant. In conclusion, our results demonstrate that in pharmacological doses the estrogen metabolites 2-ME and 2-OHE2 show inhibiting effects on the proliferation of endometriotic cells and may be promising substances for the treatment of endometriosis.


A field trial of a survey method for estimating the coverage of selective feeding programmes.
To test a survey method for estimating the coverage of selective feeding programmes in humanitarian emergencies. The trial survey used a stratified design with strata that were defined using the centric systematic area sample method. Thirty 100 km2 quadrats were sampled. The communities located closest to the centre of each quadrat were sampled using a case-finding approach. The method proved simple and rapid to implement and allowed overall and per-quadrat coverage to be estimated. Overall coverage was 20.0% (95% confidence intervals, 13.8-26.3%). Per-quadrat coverage ranged from zero (in nine quadrats) to 50% (in one quadrat). Coverage was highest in the quadrats closest to therapeutic feeding centres and in quadrats containing major roads leading to the towns in which therapeutic feeding centres were located. The method should be used, in preference to WHO Expanded Programme on Immunization (EPI)-derived survey methods, for estimating the coverage of selective feeding programmes. Its use should also be considered when evaluating the coverage of other selective entry programmes or when coverage is likely to be spatially inhomogeneous.


Bradykinin receptor gene variant and human physical performance.
Accumulating evidence suggests that athletic performance is strongly influenced by genetic variation. One such locus of influence is the gene for angiotensin-I converting enzyme (ACE), which exhibits a common variant [ACE insertion (I)/deletion (D)]. ACE can drive formation of vasoconstrictor ANG II but preferentially degrades vasodilator bradykinin. The ACE I allele is associated with higher kinin activity. A common gene variant in the kinin beta(2) receptor (B(2)R) exists: the -9 as opposed to +9 allele is associated with higher receptor mRNA expression. We tested whether this variant was associated with the efficiency of muscular contraction [delta efficiency (DE)] in 115 healthy men and women, or with running distance among 81 Olympic standard track athletes. We further sought evidence of biological interaction with ACE I/D genotype. DE was highly significantly associated with B(2)R genotype (23.84 +/- 2.41 vs. 24.25 +/- 2.81 vs. 26.05 +/- 2.26% for those of +9/+9 vs. +9/-9 vs. -9/-9 genotype; n = 25, 61, and 29, respectively; P = 0.0008 for ANOVA adjusted for sex). There was evidence for interaction with ACE I/D genotype, with individuals who were ACE II, with B(2)R -9/-9 having the highest DE at baseline. The ACE I/B(2)R -9 "high kinin receptor activity" haplotype was significantly associated with endurance (predominantly aerobic) event among elite athletes (P = 0.003). These data suggest that common genetic variation in the B(2)R is associated with efficiency of skeletal muscle contraction and with distance event of elite track athletes and that at least part of the associations of ACE and fitness phenotypes is through elevation of kinin activity.


Mechanisms whereby insulin increases diacylglycerol in BC3H-1 myocytes.
We previously suggested that insulin increases diacylglycerol (DAG) in BC3H-1 myocytes, both by increases in synthesis de novo of phosphatidic acid (PA) and by hydrolysis of non-inositol-containing phospholipids, such as phosphatidylcholine (PC) and phosphatidylethanolamine (PE). We have now evaluated these insulin effects more thoroughly, and several potential mechanisms for their induction. In studies of the effect on PA synthesis de novo, insulin stimulated [2-3H]glycerol incorporation into PA, DAG, PC/PE and total glycerolipids of BC3H-1 myocytes, regardless of whether insulin was added simultaneously with, or after 2 h or 3 or 10 days of prelabelling with, [2-3H]glycerol. In prelabelled cells, time-related changes in [2-3H]glycerol labelling of DAG correlated well with increases in DAG content: both were maximal in 30-60 s and persisted for 20-30 min. [2-3H]Glycerol labelling of glycerol 3-phosphate, on the other hand, was decreased by insulin, presumably reflecting increased utilization for PA synthesis. Glycerol 3-phosphate concentrations were 0.36 and 0.38 mM before and 1 min after insulin treatment, and insulin effects could not be explained by increases in glycerol 3-phosphate specific radioactivity. In addition to that of [2-3H]glycerol, insulin increased [U-14C]glucose and [1,2,3-3H]glycerol incorporation into DAG and other glycerolipids. Effects of insulin on [2-3H]glycerol incorporation into DAG and other glycerolipids were half-maximal and maximal at 2 nM- and 20 nM-insulin respectively, and were not dependent on glucose concentration in the medium, extracellular Ca2+ or protein synthesis. Despite good correlation between [3H]DAG and DAG content, calculated increases in DAG content from glycerol 3-phosphate specific radioactivity (i.e. via the pathway of PA synthesis de novo) could account for only 15-30% of the observed increases in DAG content. In addition to increases in [3H]glycerol labelling of PC/PE, insulin rapidly (within 30 s) increased PC/PE labelling by [3H]arachidonic acid, [3H]myristic acid, and [14C]choline. Phenylephrine, ionophore A23187 and phorbol esters did not increase [2-3H]glycerol incorporation into DAG or other glycerolipids in 2-h-prelabelling experiments; thus activation of the phospholipase C which hydrolyses phosphatidylinositol, its mono- and bis-phosphate, Ca2+ mobilization, and protein kinase C activation, appear to be ruled out as mechanisms to explain the insulin effect on synthesis de novo of PA, DAG and PC.(ABSTRACT TRUNCATED AT 400 WORDS)


Less Intense Postacute Care, Better Outcomes For Enrollees In Medicare Advantage Than Those In Fee-For-Service.
Traditional fee-for-service (FFS) Medicare's prospective payment systems for postacute care provide little incentive to coordinate care or control costs. In contrast, Medicare Advantage plans pay for postacute care out of monthly capitated payments and thus have stronger incentives to use it efficiently. We compared the use of postacute care in skilled nursing and inpatient rehabilitation facilities by enrollees in Medicare Advantage and FFS Medicare after hospital discharge for three high-volume conditions: lower extremity joint replacement, stroke, and heart failure. After accounting for differences in patient characteristics at discharge, we found lower intensity of postacute care for Medicare Advantage patients compared to FFS Medicare patients discharged from the same hospital, across all three conditions. Medicare Advantage patients also exhibited better outcomes than their FFS Medicare counterparts, including lower rates of hospital readmission and higher rates of return to the community. These findings suggest that payment reforms such as bundling in FFS Medicare may reduce the intensity of postacute care without adversely affecting patient health.


Crystal structure of the 14-3-3zeta:serotonin N-acetyltransferase complex. a role for scaffolding in enzyme regulation.
Serotonin N-acetyltransferase (AANAT) controls the daily rhythm in melatonin synthesis. When isolated from tissue, AANAT copurifies with isoforms epsilon and zeta of 14-3-3. We have determined the structure of AANAT bound to 14-3-3zeta, an association that is phosphorylation dependent. AANAT is bound in the central channel of the 14-3-3zeta dimer, and is held in place by extensive interactions both with the amphipathic phosphopeptide binding groove of 14-3-3zeta and with other parts of the central channel. Thermodynamic and activity measurements, together with crystallographic analysis, indicate that binding of AANAT by 14-3-3zeta modulates AANAT's activity and affinity for its substrates by stabilizing a region of AANAT involved in substrate binding.