Modeling HIV-1 Latency Using Primary CD4+ T Cells from Virally Suppressed HIV-1-Infected Individuals on Antiretroviral Therapy.
The low frequency of latently HIV-infected cells in vivo limits the testing of potential HIV cure strategies using cells from successfully suppressed individuals. To date, primary cell models of latency use cells infected in vitro Primary CD4+ T cell models carrying an individual's endogenous HIV reservoir that recapitulate in vivo conditions of HIV latency are still outstanding. We developed a primary CD4+ T cell model of HIV latency derived from memory CD4+ T cells isolated from virally suppressed HIV-infected individuals that recapitulates HIV-1 latency and viral reactivation events. This model is based on the expansion of primary CD4+ T cells up to 300-fold in cell number. These cells reestablish a resting state without active virus production after extended culture and maintain a stable number of total HIV proviruses. The ability of these cells to respond to various classes of latency-reversing agents is similar to that of ex vivo CD4+ T cells directly isolated from blood. Importantly, viral outgrowth assays confirmed the ability of these expanded cells to produce replication-competent endogenous virus. In sum, this model recapitulates ex vivo viral reactivation conditions, captures the variability between individuals with different HIV reservoirs, and provides large numbers of cells for testing multiple agents from a single donor. The use of this novel model will allow accurate exploration of novel cure approaches aimed either at promoting viral reactivation or maintaining sustained latency.IMPORTANCE Primary cell models of HIV latency have been very useful to identify mechanisms contributing to HIV latency and to evaluate potential HIV cure strategies. However, the current models utilize in vitro infection with exogenous virus that does not fully recapitulate virus reactivation profiles of endogenous HIV in in vivo-infected CD4+ T cells. In contrast, obtaining sufficient amounts of CD4+ T cells from HIV-infected individuals to interrogate the HIV reservoir in vitro requires leukapheresis. In the model we propose here, in vitro expansion and extended culture of primary CD4+ T cells isolated from virally suppressed HIV-infected individuals enable obtaining large numbers of cells harboring endogenous latent HIV reservoirs without performing leukapheresis. This model captures the variability of HIV reservoirs seeded in different individuals and should be useful to evaluate future HIV cure strategies.


Are cancer-related decision aids effective? A systematic review and meta-analysis.
Decision aids (DAs) have been developed to improve communication between health professionals and patients, and to involve patients in decisions about their health care. Cancer-related decisions can be difficult due to problems in communicating complex information about prognosis and the modest benefits of available treatments. We conducted a systematic review of cancer-related DAs. Randomized controlled trials (RCTs) of cancer-related DAs about screening, prevention, and treatment decision making were included. We completed a comprehensive literature search and conducted both qualitative and quantitative analyses. We also conducted a meta regression to explore heterogeneity of effect estimates. We identified 34 RCTs of DAs in a screening (n = 22 trials) or preventive/treatment (n = 12 trials) context. DAs significantly improved knowledge about screening options when compared to usual practice (weighted average effect size, 0.50; 95% CI, 0.27 to 0.73; P < .0001). A similar effect on knowledge was also found for preventive/treatment options (weighted average effect size, 0.50; 95% CI, 0.31 to 0.70; P < .0001). Overall, general anxiety was not increased in most trials and was significantly reduced in a screening context. Decisional conflict was reduced overall but not when screening and preventive/treatment studies were analyzed separately. There were few differences between different types of DAs. Cancer-related DAs are effective in increasing patient knowledge compared with usual practice without increasing anxiety particularly in the area of cancer screening. Further research is needed to determine the effectiveness of DAs in the prevention and treatment context.


Negative biotic interactions drive predictions of distributions for species from a grassland community.
Understanding the factors that determine species' geographical distributions is important for addressing a wide range of biological questions, including where species will be able to maintain populations following environmental change. New methods for modelling species distributions include the effects of biotic interactions alongside more commonly used abiotic variables such as temperature and precipitation; however, it is not clear which types of interspecific relationship contribute to shaping species distributions and should therefore be prioritized in models. Even if some interactions are known to be influential at local spatial scales, there is no guarantee they will have similar impacts at macroecological scales. Here we apply a novel method based on information theory to determine which types of interspecific relationship drive species distributions. Our results show that negative biotic interactions such as competition have the greatest effect on model predictions for species from a California grassland community. This knowledge will help focus data collection and improve model predictions for identifying at-risk species. Furthermore, our methodological approach is applicable to any kind of species distribution model that can be specified with and without interspecific relationships.


Analysis of a novel gene, Sdgc, reveals sex chromosome-dependent differences of medaka germ cells prior to gonad formation.
In vertebrates that have been examined to date, the sexual identity of germ cells is determined by the sex of gonadal somatic cells. In the teleost fish medaka, a sex-determination gene on the Y chromosome, DMY/dmrt1bY, is expressed in gonadal somatic cells and regulates the sexual identity of germ cells. Here, we report a novel mechanism by which sex chromosomes cell-autonomously confer sexually different characters upon germ cells prior to gonad formation in a genetically sex-determined species. We have identified a novel gene, Sdgc (sex chromosome-dependent differential expression in germ cells), whose transcripts are highly enriched in early XY germ cells. Chimeric analysis revealed that sexually different expression of Sdgc is controlled in a germ cell-autonomous manner by the number of Y chromosomes. Unexpectedly, DMY/dmrt1bY was expressed in germ cells prior to gonad formation, but knockdown and overexpression of DMY/dmrt1bY did not affect Sdgc expression. We also found that XX and XY germ cells isolated before the onset of DMY/dmrt1bY expression in gonadal somatic cells behaved differently in vitro and were affected by Sdgc. Sdgc maps close to the sex-determination locus, and recombination around the two loci appears to be repressed. Our results provide important insights into the acquisition and plasticity of sexual differences at the cellular level even prior to the developmental stage of sex determination.


International committee for monitoring assisted reproductive technologies: world report on assisted reproductive technologies, 2007.
To analyze information on assisted reproductive technology (ART) performed worldwide, and trends in outcomes over successive years. Cross-sectional survey on access, efficiency, and safety of ART procedures performed in 55 countries during 2007. Not applicable. Infertile women and men undergoing ART globally. Collection and analysis of international ART data. Number of cycles performed, by country and region, including pregnancies, single and multiple birth rates, and perinatal mortality. Overall, >1,251,881 procedures with ART were reported, and resulted in 229,442 reported babies born. The availability of ART varied by country, from 12 to 4,140 treatments per million population. Of all aspiration cycles, 65.2% (400,617 of 614,540) were intracytoplasmic sperm injection. The overall delivery rate per fresh aspiration was 20.3%, and for frozen-embryo transfer (FET), 18.4%, with a cumulative delivery rate of 25.8%. With wide regional variations, single-embryo transfer represented 23.4% of fresh transfers, and the proportion of deliveries with twins and triplets from fresh transfers was 22.3% and 1.2%, respectively. The perinatal mortality rate was 19.9 per 1,000 births for fresh in vitro fertilization using intracytoplasmic sperm injection, and 9.6 per 1,000 for FET. The proportion of women aged ≥40 years increased to 19.8% from 15.5% in 2006. The international trend toward <3 transferred embryos continued, as did the wider uptake of FET. This was achieved without compromising delivery rates. The application of ART for women aged >40 years was a major component of ART services in some regions and countries.


The reaction-limited kinetics of membrane-to-surface adhesion and detachment.
Biological adhesion is frequently mediated by specific membrane proteins (adhesion molecules). Starting with the notion of adhesion molecules, we present a simple model of the physics of membrane-to-surface attachment and detachment. This model consists of coupling the equations for deformation of an elastic membrane with equations for the chemical kinetics of the adhesion molecules. We propose a set of constitutive laws relating bond stress to bond strain and also relating the chemical rate constants of the adhesion molecules to bond strain. We derive an exact formula for the critical tension. We also describe a fast and accurate finite difference algorithm for generating numerical solutions of our model. Using this algorithm, we are able to compute the transient behaviour during the initial phases of adhesion and detachment as well as the steady-state geometry of adhesion and the velocity of the contact. An unexpected consequence of our model is the predicted occurrence of states in which adhesion cannot be reversed by application of tension. Such states occur only if the adhesion molecules have certain constitutive properties (catch-bonds). We discuss the rational for such catch-bonds and their possible biological significance. Finally, by analysis of numerical solutions, we derive an accurate and general expression for the steady-state velocity of attachment and detachment. As applications of the theory, we discuss data on the rolling velocity of granulocytes in post-capillary venules and data on lectin-mediated adhesion of red cells.


Proteins dominate in the surface layers formed on materials exposed to extracellular polymeric substances from bacterial cultures.
The chemical compositions of the surface conditioning layers formed by different types of solutions (from isolated EPS to whole culture media), involving different bacterial strains relevant for biocorrosion were compared, as they may influence the initial step in biofilm formation. Different substrata (polystyrene, glass, steel) were conditioned and analyzed by X-ray photoelectron spectroscopy. Peak decomposition and assignment were validated by correlations between independent spectral data and the ubiquitous presence of organic contaminants on inorganic substrata was taken into account. Proteins or peptides were found to be a major constituent of all conditioning layers and polysaccharides were not present in appreciable concentrations; the proportion of nitrogen which may be due to DNA was lower than 15%. There was no significant difference between the compositions of the adlayers formed from different conditioning solutions, except for the adlayers produced with tightly bound EPS extracted from D. alaskensis.


Gestational trophoblastic diseases: 4. Presentation with persistent low positive human chorionic gonadotropin test results.
A high proportion of women with persistent low levels of hCG, in the absence of pregnancy or any evidence of tumor, have received chemotherapy and hysterectomy for assumed malignancy. Such chemotherapy and surgery were ineffective and unwarranted. This study identifies the causes of persistent low level of hCG and provides guidelines for the management of these patients, preventing unnecessary treatment in the future. The USA hCG Reference Service has consulted on 170 women with low levels of hCG persisting for 3 months or longer. Serum total hCG was measured in the Diagnostic Products Corporation (DPC) Immulite assay and hyperglycosylated hCG in the Nichols Advantage test. Among these 170 patients, the average persistent hCG result was 102 +/- 152 mIU/ml, with a range of 6.1-900 mIU/ml. Thirteen (7.6%) of the 170 patients had true malignancy, 5 had placental site trophoblastic tumor, 3 had other gestational trophoblastic neoplasms (GTN), and 5 had non-trophoblastic malignancies. The remaining 157 patients had false-positive hCG, quiescent gestational trophoblastic disease (quiescent GTD), or pituitary hCG (hCG of pituitary origin). Of 71 patients with false-positive hCG, 47 patients received chemotherapy and 9 had surgery that had no effect on the level of hCG. Five of these patients with false-positive hCG were being monitored for hydatidiform mole or GTN. The majority of these cases were first investigated following an incidental pregnancy test. Of 69 patients who had quiescent GTD, 41 received chemotherapy and 9 underwent hysterectomy. All these therapies were unnecessary and ineffective. While 21 patients with quiescent GTD followed incidental pregnancy tests, the majority were discovered while monitoring patients after treatment for hydatidiform mole or GTN/choriocarcinoma (n = 48). Seventeen cases of pituitary hCG were found among those women who were peri- or post-menopause. Two patients also received chemotherapy for assumed malignancy which was not present. Clinicians frequently assume that an elevated hCG implies that a patient is pregnant or has GTD or recurrent GTN, even when apart from the pregnancy test, no clinical evidence was found to support such a diagnosis. In most of these cases of persistent low hCG etiologies, all therapies were found unnecessary and ineffective. Guidelines are proposed for managing these patients. It is essential to demonstrate a malignancy clinically and with readily available biochemical tests before initiating therapy. This applies whether the patient is identified by an incidental pregnancy test or is actively being monitored for gestational trophoblastic disease.


The signal recognition particle (SRP) RNA links conformational changes in the SRP to protein targeting.
The RNA component of the signal recognition particle (SRP) is universally required for cotranslational protein targeting. Biochemical studies have shown that SRP RNA participates in the central step of protein targeting by catalyzing the interaction of the SRP with the SRP receptor (SR). SRP RNA also accelerates GTP hydrolysis in the SRP.SR complex once formed. Using a reverse-genetic and biochemical analysis, we identified mutations in the E. coli SRP protein, Ffh, that abrogate the activity of the SRP RNA and cause corresponding targeting defects in vivo. The mutations in Ffh that disrupt SRP RNA activity map to regions that undergo dramatic conformational changes during the targeting reaction, suggesting that the activity of the SRP RNA is linked to the major conformational changes in the signal sequence-binding subunit of the SRP. In this way, the SRP RNA may coordinate the interaction of the SRP and the SR with ribosome recruitment and transfer to the translocon, explaining why the SRP RNA is an indispensable component of the protein targeting machinery.


Effects of tonin on vascular smooth muscle of the hypertensive rat and normal rabbit.
1. The response of arterial smooth muscle to noradrenaline was studied in one-clip hypertensive rats with or without the contralateral/kidney and in normotensive rabbits. 2. Strips of aorta from one-kidney, one-clip hypertensive animals were less responsive to noradrenaline than normotensive control rats. The contractile response of strips from two-kidney, one-clip hypertensive animals was not different from the control group. These results suggest that the mechanisms responsible for the lesser reactivity in the one-kidney hypertensive group are not a consequence of elevated blood pressure itself, but may be related to the intrinsic contractility of aortic smooth muscle. 3. Tonin potentiated the contraction induced by noradrenaline in aortic strips from hypertensive and normotensive rats. However, this effect was more important in the one-kidney, one-clip hypertensive animals. In the aortic and mesenteric strips from normal rabbits, tonin produced not only potentiation to noradrenaline but direct contraction. 4. The potentiation to noradrenaline and the direct effect of tonin were not affected by a variety of antagonists but were blocked by a calcium ion antagonist, verapamil, suggesting that tonin may act directly on vascular smooth muscle through mechanisms which might be mediated by calcium ions.


Role of nuclear cardiology in advancing cardiac surgery.
Cardiac surgeons are commonly faced with issues regarding the balance between the potential risk and the potential benefit of a surgical procedure. Nuclear cardiology procedures [myocardial perfusion SPECT (MPS) and positron emission tomography (PET)] provide the surgeon with objective information that augments standard clinical and angiographic assessments with respect to diagnosis, prognosis, and potential benefit from intervention. Development of the technology and methodology of gated MPS acquisition and interpretation allows assessment of the extent and severity of hypoperfused but viable myocardium, as well as global LVEF and LV volume measurements, diastolic function, and LV geometry. With PET, myocardial metabolism and blood flow reserve can also be measured. This chapter provides insight into the current evidence regarding settings in which nuclear cardiology procedures are helpful to the surgeon in assessment of patients having or being considered for cardiac surgery in the setting of coronary artery disease (CAD). Overall, a risk-benefit approach to MPS results is proposed, with principal focus on identifying patients at risk for major cardiac events who may benefit from a surgical procedure.


Increased diversity of egg-associated bacteria on brown trout (Salmo trutta) at elevated temperatures.
The taxonomic composition of egg-associated microbial communities can play a crucial role in the development of fish embryos. In response, hosts increasingly influence the composition of their associated microbial communities during embryogenesis, as concluded from recent field studies and laboratory experiments. However, little is known about the taxonomic composition and the diversity of egg-associated microbial communities within ecosystems; e.g., river networks. We sampled late embryonic stages of naturally spawned brown trout at nine locations within two different river networks and applied 16S rRNA pyrosequencing to describe their bacterial communities. We found no evidence for a significant isolation-by-distance effect on the composition of bacterial communities, and no association between neutral genetic divergence of fish host (based on 11 microsatellites) and phylogenetic distances of the composition of their associated bacterial communities. We characterized core bacterial communities on brown trout eggs and compared them to corresponding water samples with regard to bacterial composition and its presumptive function. Bacterial diversity was positively correlated with water temperature at the spawning locations. We discuss this finding in the context of the increased water temperatures that have been recorded during the last 25 years in the study area.


Isolation of a novel inducible rat heat-shock protein (HSP70) gene and its expression during ischaemia/hypoxia and heat shock.
Most of the members of the mammalian heat-shock protein (HSP) gene family have been studied and isolated from human and mouse cells. Few studies have concentrated on the HSPs of rat, a commonly used experimental animal. We have isolated and characterized a novel inducible rat HSP70 gene using an HSP70 cDNA sequence obtained from an ischaemic rat heart cDNA library. The isolated rat HSP70 gene was found to be a functional gene, as indicated by RNAase-protection and Northern-blot analysis. The deduced amino acid sequence of the inducible rat HSP70 exhibits a high degree of similarity to previously isolated mammalian inducible HSP70 gene products. Expression of the inducible HSP70 gene in rat myogenic cells (H9c2) is markedly increased after relatively short periods of hypoxia as well as by heat shock. Two heat-shock elements (HSE) are present in the rat HSP70 promoter. Transient transfection of rat HSP70 promoter/chloramphenicol acetyltransferase constructs into H9c2 cells shows that the presence of either of the two HSEs is sufficient for heat-shock inducibility. In contrast, induction of the rat HSP70/chloramphenicol acetyltransferase constructs by hypoxia is only detectable when both HSEs are present. This leads us to conclude that the induction of HSP70 by hypoxia and heat shock occurs through the same regulatory HSEs but the activation of the inducible HSP70 gene by heat shock is several-fold higher than by hypoxia.


Activated type I TGFbeta receptor kinase enhances the survival of mammary epithelial cells and accelerates tumor progression.
We have examined the effects of transforming growth factor-beta (TGFbeta) signaling on mammary epithelial cell survival. Transgenic mice expressing an active mutant of Alk5 in the mammary gland (MMTV-Alk5(T204D)) exhibited reduced apoptosis in terminal endbuds and during postlactational involution. Transgene-expressing mammary cells contained lower Smad2/3 and higher c-myc levels than controls, high ligand-independent phosphatidylinositol-3 kinase (PI3K) and Akt activities, and were insensitive to TGFbeta-mediated growth arrest. Treatment with a proteasome inhibitor increased Smad2/3 levels and ligand-independent Smad transcriptional reporter activity, as well as reduced both c-myc protein and basal cell proliferation. Treatment with an Alk5 kinase small-molecule inhibitor upregulated Smad2/3 levels, reduced PI3K activity, P-Akt, and c-myc, and inhibited cell survival. Although Alk5(T204D)-expressing mice did not develop mammary tumors, bigenic MMTV-Alk(T204D) x Neu mice developed cancers that were more metastatic than those occurring in MMTV-Neu transgenics. These data suggest that (1) TGFbeta can signal to PI3K/Akt and enhance mammary epithelial cell survival in vivo before cytological or histological evidence of transformation, and (2) TGFbeta signaling can provide epithelial cells with a 'gain-of-function' effect that synergizes with oncogene-induced transformation.


The contralateral transfalcine transprecuneus approach to the atrium of the lateral ventricle: operative technique and surgical results.
Surgical approaches to the atrium of the lateral ventricle remain a challenging neurosurgical issue because of the eloquent nature of the surrounding anatomy. To report our operative techniques and preliminary surgical results with the contralateral transfalcine transprecuneus approach. A retrospective data review was performed of patients undergoing a contralateral transfalcine transprecuneus approach for the resection of lesions in the atrium of the lateral ventricle. Patients were positioned in the prone position with a 30° elevation, and a 15° rotation was used. After a contralateral parasagittal parieto-occipital craniotomy and falx incision, the corticotomy in the contralateral precuneus gyrus created a corridor to the tumor. An endoscope was used to assist with the surgery. Headache was the primary preoperative symptom, which improved in all patients after surgery. After treatment, symptoms were improved in all 3 patients with hemiparesis and in 3 of 6 patients with preexisting visual deficits; symptoms were unchanged in the other 3 patients with visual deficits during the 13- to 38-month follow-up. Nine lesions were totally removed, and 1 metastatic breast cancer lesion was subtotally removed; all patients had good neurological outcomes and no operative mortality. The contralateral transfalcine transprecuneus approach is appropriate for most lesions in the atrium of the lateral ventricle. It provides a wider surgical angle (especially for the lateral extension) and reduces the risk of disturbance of the optic radiation compared with the conventional approaches. The use of magnetic resonance venography-magnetic resonance imaging neuronavigation makes the procedure much easier and more accurate, and the neuroendoscope adds to the visualization of the microscope and can reduce surgical complications.


Type 2 diabetes increases and metformin reduces total, colorectal, liver and pancreatic cancer incidences in Taiwanese: a representative population prospective cohort study of 800,000 individuals.
Metformin protection against cancer risk in Orientals is uncertain. We examined the possible metformin effect on total, esophageal, gastric, colorectal (CRC), hepatocellular (HCC) and pancreatic cancers in a Taiwanese cohort. A representative sample of 800,000 was drawn from the Taiwanese National Health Insurance data of 2000. A cohort of 480,984 participants 20 years or older, diabetes-cancer-free on 1st January 2000 was formed and categorized as four groups by DM and metformin usage status. Eligible incident cancer events had to occur one year after the index date until the end of 2007. The Cox proportional-hazards model evaluated relative risk of cancer for treated DM patients with or without metformin. The covariates included age, gender, other oral anti-hyperglycemic medication, Charlson comorbidity index (CCI) score and metformin exposure dosage and duration. With diabetes but no anti-hyperglycemic medication, cancer incidence density increased at least 2-fold for total, CRC and HCC. On metformin, total, CRC and HCC incidences decreased to near non-diabetic levels but to varying degrees depending on gender and cancer type (CRC in women, liver in men). Adjustment for other oral anti-hyperglycemic agents usage and CCI made the benefit of metformin more evident [hazard ratios (95% confidence intervals): total 0.12 (0.08-0.19), CRC 0.36 (0.13-0.98), liver 0.06 (0.02-0.16), pancreas 0.15 (0.03-0.79)]. There was a significant gender interaction with metformin in CRC which favored women. Metformin dosage for a significant decrease in cancer incidence was ≤ 500 mg/day. Metformin can reduce the incidences of several gastroenterological cancers in treated diabetes.


Ex vivo phenotype and frequency of influenza virus-specific CD4 memory T cells.
Recent advances in class II tetramer staining technology have allowed reliable direct ex vivo visualization of antigen-specific CD4 T cells. In order to define the frequency and phenotype of a prototype response to a nonpersistent pathogen, we have used such techniques to analyze influenza virus-specific memory CD4 T cells directly from blood. These responses are stably detectable ex vivo at low frequencies (range, 0.00012 to 0.0061% of CD4 T cells) and display a distinct "central memory" CD62L(+) phenotype.


Towards a universal concept of vulnerability: Broadening the evidence from the elderly to perinatal health using a Delphi approach.
The concept 'vulnerability' is prevalent in the public domain, health care, social institutions and multidisciplinary research. Conceptual heterogeneity is present, hampering the creation of a common evidence-base of research achievements and successful policies. Recently an international expert group combined a specific literature review with a 2-stage Delphi procedure, arriving at a seemingly universal concept of vulnerability for the elderly with applications for research instruments. We replicated and extended this study, to generalize this result to health in general, and perinatal health in particular. Two independent expert panels (general health, perinatal health) repeated the Delphi-procedure, using an extended and updated literature review to derive statements on the concept and defining pathways of vulnerability. Additional views were collected on research tools. Consensus-by-design was explicitly avoided. Data collection and processing was independent. Both panels showed surprising convergence on the pathways of vulnerability to health/ill-health, and their interaction. The agreed conceptual model describes a dynamic relation between health and ill-health and vulnerability. The 2 key pathways that link to vulnerability, are complementary, but not symmetrical as biological processes of maintaining health or obtaining better health are not reciprocal to recovery, so also not in terms of vulnerability impacts. An individual's degree of vulnerability is the net balance of risk effects and protective and healing factors (socially, biologically and in terms of health literacy and health care access). These factors can for measurement purposes (according to the panels: interview for exploration, checklists for population research) be grouped into 'material resources', 'taking responsibility for one's own health', 'risky activities and behaviors', and 'social support'. Supportive and transforming action can thus be undertaken. A universal concept of vulnerability in the context of health was successfully derived after careful replication and extension of an international Delphi study on vulnerability among the elderly.


Distinct cerebellar contributions to intrinsic connectivity networks.
Convergent data from various scientific approaches strongly implicate cerebellar systems in nonmotor functions. The functional anatomy of these systems has been pieced together from disparate sources, such as animal studies, lesion studies in humans, and structural and functional imaging studies in humans. To better define this distinct functional anatomy, in the current study we delineate the role of the cerebellum in several nonmotor systems simultaneously and in the same subjects using resting state functional connectivity MRI. Independent component analysis was applied to resting state data from two independent datasets to identify common cerebellar contributions to several previously identified intrinsic connectivity networks (ICNs) involved in executive control, episodic memory/self-reflection, salience detection, and sensorimotor function. We found distinct cerebellar contributions to each of these ICNs. The neocerebellum participates in (1) the right and left executive control networks (especially crus I and II), (2) the salience network (lobule VI), and (3) the default-mode network (lobule IX). Little to no overlap was detected between these cerebellar regions and the sensorimotor cerebellum (lobules V-VI). Clusters were also located in pontine and dentate nuclei, prominent points of convergence for cerebellar input and output, respectively. The results suggest that the most phylogenetically recent part of the cerebellum, particularly crus I and II, make contributions to parallel cortico-cerebellar loops involved in executive control, salience detection, and episodic memory/self-reflection. The largest portions of the neocerebellum take part in the executive control network implicated in higher cognitive functions such as working memory.


Practical GC/MS analysis of oxidation dye components in hair fiber as a forensic investigative procedure.
The purpose of this study was to improve the reliability of discrimination (or identification) of dyed hair by analyzing chemical substances present in the hair, as an addition to the conventional macroscopical and microscopical examinations and ABO blood group examination. Oxidation hair-dye components such as p-phenylenediamine (PPDA), toluylene-2,5-diamine (T-2,5-DA), o-aminophenol (OAP), m-aminophenol (MAP), p-aminophenol (PAP) and p-amino-o-cresol (PAOC) were selected as the object of study. After alkaline-digestion, hair samples were adjusted to a pH of 12.6 to 12.8, and applied onto an Extrelut column. After 15 min, the components were simultaneously extracted and derivatized with n-hexane including 1% heptafluoro-n-butyryl (HFB) chloride. Their HFB derivatives within a condensed sample were diluted in ethyl acetate, and analyzed by gas chromatography-mass spectrometry (GC-MS) with full mass scanning or selected ion monitoring. For estimating their levels, 2,4,6-trimethylaniline was used as the internal standard. Standard curves obtained by preparing a 5 cm segment of control hair spiked with authentic substances were linear, ranging from 0.1 to 4.0 micrograms for PPDA and T-2,5-DA, and from 0.01 to 0.4 microgram for OAP, MAP, PAP and PAOC. The coefficient of variation of inter-day precisions (each n = 4) was below 16% for PPDA, below 20% for OAP and PAP and below 24% for T-2,5-DA, MAP and PAOC. These components were detectable even at 6 ng for PPDA, T-2,5-DA, MAP and PAP, 8 ng for OAP, and 4 ng for PAOC. Recovery percents using this procedure ranged from 54 to 86%. By using actual dyed hair samples this method was shown to provide high sensitivity for accurate detection.


Factors predicting incidence of post-operative delirium in older people following hip fracture surgery: a systematic review and meta-analysis.
Delirium is one of the most common complications following hip fracture surgery in older people. This study identified pre- and peri-operative factors associated with the development of post-operative delirium following hip fracture surgery. Published and unpublished literature were searched to identify all evidence reporting variables on patient characteristics, on-admission, intra-operative and post-operative management assessing incident delirium in older people following hip fracture surgery. Pooled odds ratio (OR) and mean difference of those who experienced delirium compared to those who did not were calculated for each variable. Evidence was assessed using the Downs and Black appraisal tool and interpreted using the GRADE approach. A total of 6704 people (2090 people with post-operative delirium) from 32 studies were analysed. There was moderate evidence of nearly a two-times greater probability of post-operative delirium for those aged 80 years and over (OR: 1.77; 95% CI: 1.09, 2.87), whether patients lived in a care institution pre-admission (OR: 2.65; 95% CI: 1.79, 3.92), and a six-times greater probability of developing post-operative delirium with a pre-admission diagnosis of dementia (OR: 6.07, 95% CI: 4.84, 7.62). There was no association with intra-operative variables and probability of delirium. Clinicians treating people with a hip fracture should be vigilant towards post-operative delirium if their patients are older, have pre-existing cognitive impairment and poorer overall general health. This is also the case for those who experience post-operative complications such as pneumonia or a urinary tract infection. Copyright © 2017 John Wiley & Sons, Ltd.


Rapid Analysis of Anabolic Steroid Metabolites in Urine by Combining Field Asymmetric Waveform Ion Mobility Spectrometry with Liquid Chromatography and Mass Spectrometry.
The combination of field asymmetric waveform ion mobility spectrometry with liquid chromatography-mass spectrometry (LC-FAIMS-MS) has been developed for the analysis of glucuronide and sulfate metabolites of seven anabolic-androgenic steroids in urine. Separation by FAIMS-MS was investigated in positive ion mode for selected cationic adducts (H+, NH4+, Na+, K+, and Cs+). LC-FAIMS-MS analysis of the doubly sodiated adducts ([M + 2Na - H]+) of isobaric and coeluting steroid metabolites allowed their rapid (8 min) qualitative and quantitative determination in spiked urine using hydrophilic interaction liquid chromatography prior to FAIMS-MS separation, with discrimination >95% achieved between the steroids investigated. A quantitative evaluation of the LC-FAIMS-MS method was performed giving limits of detection in the range 1-6 ng mL-1, limits of quantification in the range 3-20 ng mL-1, with reproducibility (%RSD < 10%; n = 6) and linearity (R2 > 0.99). The LC-FAIMS-MS method demonstrates increases in signal-to-noise ratios for the doubly sodiated steroid metabolites in unspiked urine (>250%) by the reduction of isobaric interferences from the matrix. An alternative or additional tool for identification of the steroid metabolites is based on the observations of different patterns of sodium acetate clusters that are characteristic for each metabolite.


An introduction to standardized clinical nomenclature for dysmorphic features: the Elements of Morphology project.
Human structural malformations (anomalies or birth defects) have an enormous and complex range of manifestations and severity. The description of these findings can be challenging because the variation of many of the features is continuous and only some of them can be objectively assessed (that is, measured), among other factors. An international group of clinicians resolved to develop a set of terms that could be used to describe human structural malformations, under the general project name 'Elements of Morphology'. Here, the background to the project, progress to date, and the practical implementation of the terminology in research reporting is discussed.


Caregiving-related needs analysis: a proposed model reflecting current research and socio-political developments.
The present authors have developed a model for assessing caregiving-related problems and needs, the Economic, Social, Psychological and Educational Requirements Table (ESPERT). The model was based on an analysis of the international literature concerning caregiving-related problems, and current specific European and Italian legislation, as well as the authors' own research results (which also permitted them to design a specific instrument for the general assessment of caregivers). This paper describes the socio-psychological and socio-political background prompting the development of the model, the general instrument for the assessment of caregivers and an example of its use in a sample of caregivers of cancer patients. In the example study, 80 caregivers of cancer patients were asked to complete questionnaires evaluating their state and trait anxiety, depressive symptoms, personality traits, and specifically, their neuroticism or emotional instability, caregiving strain and the degree of satisfaction with their lives. In addition to the specific information obtained about this group of caregivers, the present results show that the completion of the ESPERT makes it possible to identify the proportion of subjects at economic, social and emotional risk, and to decide rapidly what kind of resources/interventions can be implemented, bearing in mind the most balanced cost-benefit ratio.


A novel (99m)Tc labelling strategy for the development of silica based particles for medical applications.
The reaction of high-valent fac-{(99m)TcO3}(+) complexes with alkenes ((3 + 2)-cycloaddition) has been used to develop a novel labelling procedure for silica based particles. This strategy can help to gain more detailed in vivo data of mesoporous silica by non-invasive radioimaging in the future. By applying norbornene as a coupling unit the (3 + 2)-cycloaddition reaction proceeds stereoselectively and with high reaction rates.


Haptic Exploratory Procedures of Children and Youth with and without Cerebral Palsy.
Aims: Compare haptic exploratory procedures (EPs) and exploratory movements (EMs) of children. This study also tested the interrater reliability of a novel digital recording method. Methods: Participants were 31 children with typical development (TD) (aged 6 years 1 month to 15 years 9 months; 14 male) and 23 children with spastic unilateral cerebral palsy (CP) (aged 6 years to 15 years 5 months; 13 males; right hemiplegia, n = 12). Results: There were no statistically significant differences between groups for expected EP (p = .15), additional EPs (p = .78), or EMs (p = .69) but there was for mean duration of exploration (p < .001) and accuracy (p < .001). This suggests that although children with CP performed similar haptic EPs for each object as children with TD, they took more time and were less accurate in their identification. There was substantial agreement between the two raters' observations of expected EP, κ = .64, p < .0005. Conclusion: Children with CP performed similar haptic EPs as their TD peers. However, despite similarities, the results indicate that for children with CP manual ability was not the primary determinant of accuracy or speed of identification. This study provides evidence for a reliable method of recording haptic EPs.


Programmable high-bit-rate pattern generator with a segmented semiconductor optical amplifier.
We have demonstrated that spatial gain modulation in a segmented semiconductor optical amplifier can be converted to a temporal signal. A four-segment amplifier was used to generate digital return-to-zero patterns at 40 Gbits/s , and this technique should be readily scalable to more than 100 Gbits/s .


Untangling occupation and activity.
Activity and occupation are two core concepts of occupational therapy that are in need of differentiation. Occupation is defined here as a person's personally constructed, one-time experience within a unique context. Activity is defined as a more general, culturally shared idea about a category of action. The ways in which subjectivity and context are handled within the concepts of occupation and activity are keys to disentangling them. The proposed untangling of the two concepts into distinct definitions is congruent with their historical origins as well as with current definitional trends. Once occupation and activity are recognized as two separate and equally valuable concepts, they offer a rich set of theoretical relations for exploration. The clarity that will result from differentiating occupation and activity will enhance disciplinary discourse and research as well as enhance the intervention efficacy, moral surety, and political strength of the profession.


Research on biomodulatory effect of natural compounds.
The purpose of this study was to determine whether the extract isolated from the artichoke Cynara cardunculus L. (ECC) had antimutagenic effect and was able to enhance the therapeutic effect of cytostatic drug cis-platinum (cis-Pt). The potential antimutagenic activity of ECC was assayed by a test on sex-linked recessive lethal mutations detection in Drosophila melanogaster males treated with ethylmethane sulfonate (EMS). The possible enhancement of cytostatic/cytotoxic effect of cis-Pt by ECC was evaluated in the cell revitalization assay by measuring cell viability via Trypan blue exclusive assay using mouse leukemia cells L1210. EMS was both toxic and genotoxic in D. melanogaster males. It statistically significantly increased the frequency of sex-linked recessive lethal mutations in comparison to the negative control. Furthermore, ECC statistically significantly reduced the genotoxic effect of EMS. It acted in a desmutagenic manner via EMS inactivation. In the cell revitalization assay, ECC enhanced the cytotoxic/cytostatic effect of cis-Pt. The therapeutic potential of ECC was established on the basis of statistically significantly lowered recovery of cis-Pt pre-treated mouse leukemia cells in the presence of ECC. The results imply that the extract isolated from artichoke C. cardunculus L. has marked beneficial activities antimutagenic and therapeutic effect enhancing) and its potential biomedical application in the combination therapy of cancer and some neurodegenerative diseases may be suggested.


The Role of Curcumin in Prevention and Management of Metastatic Disease.
In the last two decades, targeted therapies have enhanced tumor patient care and treatment success, however, metastatic growth still cannot be stopped efficiently and, therefore, mortality rates remain high. Prevention strategies against formation of metastases are the most promising approach we have, however, due to lack of clinical validation studies, they have not yet entered routine clinical care. In order to smooth the way for efficient prevention, further preclinical and large clinical studies are required. In this context, the underlying molecular mechanisms and factors that lead to metastatic growth have to be explored, and potential preventive agents have to be tested. Thereby, special attention has to be paid to natural bioactive compounds which do not exert major adverse effects, like the plant-derived polyphenol Curcumin, which is known to be a powerful antitumor agent. So far, most of the preclinical studies with Curcumin have focused on its effect on inhibiting tumor cell proliferation and invasion, although, it is known that it also inhibits metastatic spread in vivo. This review discusses the preventive potential of this natural compound not only against tumor onset, but also against formation of metastases.


Expanded insecticide catabolic activity gained by a single nucleotide substitution in a bacterial carbamate hydrolase gene.
Carbofuran-mineralizing strain Novosphingobium sp. KN65.2 produces the CfdJ enzyme that converts the N-methylcarbamate insecticide to carbofuran phenol. Purified CfdJ shows a remarkably low KM towards carbofuran. Together with the carbaryl hydrolase CehA of Rhizobium sp. strain AC100, CfdJ represents a new protein family with several uncharacterized bacterial members outside the proteobacteria. Although both enzymes differ by only four amino acids, CehA does not recognize carbofuran as a substrate whereas CfdJ also hydrolyzes carbaryl. None of the CfdJ amino acids that differ from CehA were shown to be silent regarding carbofuran hydrolytic activity but one particular amino acid substitution, i.e., L152 to F152, proved crucial. CfdJ is more efficient in degrading methylcarbamate pesticides with an aromatic side chain whereas CehA is more efficient in degrading the oxime carbamate nematicide oxamyl. The presence of common flanking sequences suggest that the cfdJ gene is located on a remnant of the mobile genetic element Tnceh carrying cehA. Our results suggest that these enzymes can be acquired through horizontal gene transfer and can evolve to degrade new carbamate substrates by limited amino acid substitutions. We demonstrate that a carbaryl hydrolase can gain the additional capacity to degrade carbofuran by a single nucleotide transversion.


Synthesis and turnover of ribulose biphosphate carboxylase and of its subunits during the cell cycle of Chlamydomonas reinhardtii.
The chloroplast enzyme ribulose-1,5-bisphosphate (Ru-1,5-P2) carboxylase (EC 4.1 1.39) is made up ot two nonidentical subunits, one synthesized in the chloroplast and the other outside. Both of these subunits of the assembled enzyme are synthesized in a stepwise manner during the synchronous cell cycle of the green alga Chlamydomonas reinhardtii. The activity of this enzyme increases in the light and this increase is due to de novo protein synthesis as shown by the measurement of the amount of protein and by the pulse incorporation of radioactive arginine in the 18S enzyme peak in linear sucrose density gradients. During the dark phase of the cell cycle, there is little change in the enzymatic activity as well as in the amount of this enzyme. Pulse-labeling studies using radioactive arginine indicated that there is a slow but detectable rate of synthesis of the carboxylase and of its subunits in the dark. Ru-1,5-P2 carboxylase, prelabeled with radioactive arginine throughout the entire light period, shows a similarly slow rate of degradation in the following dark period. This slow turnover of the enzyme in the dark accounts for the steady levels of carboxylase protein and of enzymatic activity during this period. A wide variety of inhibitors of protein synthesis by 70S and 80S ribosomes abolished the incorporation of [3H]arginine into total Ru-1,5-P2 carboxylase during short-term incubation. These results suggest a tight-coordinated control of the biosynthesis of the small and large subunits of the enzyme. This stringent control is further substantiated by the finding that both subunits are synthesized in sychrony with each other, that the ratio of radioactivity of the small to the large subunit remains constant throughout the entire light-dark cycle, and that the rates of synthesis and of degradation of both subunits are similar to that of the assembled enzyme.


Estimating the effect of sulfonylurea on HbA1c in diabetes: a systematic review and meta-analysis.
Sulfonylureas are widely prescribed glucose-lowering medications for diabetes, but the extent to which they improve glycaemia is poorly documented. This systematic review evaluates how sulfonylurea treatment affects glycaemic control. Medline, EMBASE, the Cochrane Library and clinical trials registries were searched to identify double-blinded randomised controlled trials of fixed-dose sulfonylurea monotherapy or sulfonylurea added on to other glucose-lowering treatments. The primary outcome assessed was change in HbA1c, and secondary outcomes were adverse events, insulin dose and change in body weight. Thirty-one trials with a median duration of 16 weeks were included in the meta-analysis. Sulfonylurea monotherapy (nine trials) lowered HbA1c by 1.51% (17 mmol/mol) more than placebo (95% CI, 1.25, 1.78). Sulfonylureas added to oral diabetes treatment (four trials) lowered HbA1c by 1.62% (18 mmol/mol; 95% CI 1.0, 2.24) compared with the other treatment, and sulfonylurea added to insulin (17 trials) lowered HbA1c by 0.46% (6 mmol/mol; 95% CI 0.24, 0.69) and lowered insulin dose. Higher sulfonylurea doses did not reduce HbA1c more than lower doses. Sulfonylurea treatment resulted in more hypoglycaemic events (RR 2.41, 95% CI 1.41, 4.10) but did not significantly affect the number of other adverse events. Trial length, sulfonylurea type and duration of diabetes contributed to heterogeneity. Sulfonylurea monotherapy lowered HbA1c level more than previously reported, and we found no evidence that increasing sulfonylurea doses resulted in lower HbA1c. HbA1c is a surrogate endpoint, and we were unable to examine long-term endpoints in these predominately short-term trials, but sulfonylureas appear to be associated with an increased risk of hypoglycaemic events.


Dyadic coping and relationship satisfaction: A meta-analysis.
Meta-analytic methods were used to empirically determine the association between dyadic coping and relationship satisfaction. Dyadic coping is a systemic conceptualization of the processes partners use to cope with stressors, such as stress communication, individual strategies to assist the other partner cope with stress, and partners' strategies to cope together. A total of 72 independent samples from 57 reports with a combined sum of 17,856 participants were included. The aggregated standardized zero-order correlation (r) for total dyadic coping with relationship satisfaction was .45 (p=.000). Total dyadic coping strongly predicted relationship satisfaction regardless of gender, age, relationship length, education level, and nationality. Perceptions of overall dyadic coping by partner and by both partners together were stronger predictors of relationship satisfaction than perceptions of overall dyadic coping by self. Aggregated positive forms of dyadic coping were a stronger predictor of relationship satisfaction than aggregated negative forms of dyadic coping. Comparisons among dyadic coping dimensions indicated that collaborative common coping, supportive coping, and hostile/ambivalent coping were stronger predictors of relationship satisfaction than stress communication, delegated coping, protective buffering coping, and overprotection coping. Clinical implications and recommendations for future research are provided.


Simulation of infrared scattering from ice aggregates by use of a size-shape distribution of circular ice cylinders.
The scalar optical properties (extinction coefficient, mass extinction coefficient, single-scattering albedo, and asymmetry parameter) of a distribution of randomly oriented ice aggregates are simulated generally to well within 4% accuracy by use of a size-shape distribution of randomly oriented circular ice cylinders at wavelengths in the terrestrial window region. The single-scattering properties of the ice aggregates are calculated over the whole size distribution function by the finite-difference time-domain and improved geometric optics methods. The single-scattering properties of the size-shape distribution of circular ice cylinders are calculated by the T-matrix method supplemented by scattering solutions obtained from complex-angular-momentum theory. Moreover, radiative-transfer studies demonstrate that the maximum error in brightness temperature space when the size-shape distribution of circular ice cylinders is used to represent scattering from ice aggregates is only approximately 0.4 K The methodology presented should find wide applicability in remote sensing of ice cloud and parameterization of cirrus cloud scalar optical properties in climate models.


An Imbalance in TAZ and YAP Expression in Hepatocellular Carcinoma Confers Cancer Stem Cell-like Behaviors Contributing to Disease Progression.
Transcriptional coactivator with PDZ-binding motif (TAZ) and yes-associated protein (YAP) are equivalently placed downstream effectors of the Hippo pathway with oncogenic roles in human cancers. However, the expression profiles of TAZ/YAP differ depending on the cancer cell type, suggesting that these proteins have different roles during cancer progression, yet no studies have examined the biologic significance of the balance between TAZ and YAP expression levels. Here we examined the functional roles of TAZ/YAP in hepatocellular carcinoma progression. We found that TAZ, but not YAP, was predominantly expressed in HCC. TAZ knockdown under normal conditions attenuated cell growth in HCC cells; however, TAZ knockdown combined with 5-fluorouracil treatment significantly increased chemoresistance compared with control cells. Notably, TAZ knockdown induced compensatory YAP expression and was accompanied by upregulation of CD90, a HCC-specific cancer stem cell marker. Continuous treatment with 5-fluorouracil also induced YAP expression and promoted tumor formation in vivo. Conversely, double knockdown of TAZ/YAP reduced chemoresistance and tumorigenicity. Moreover, YAP knockdown aggravated HCC cell growth to a greater degree than TAZ knockdown, and YAP overexpression was strongly associated with poor prognoses in patients with HCC. Collectively, these studies demonstrate that TAZ and YAP exhibit different functional roles in cancer progression, and a shift to predominant YAP expression upon TAZ depletion conferred cancer stem cell-like properties including chemoresistance and tumorigenicity in HCC. Therefore, targeting of both TAZ/YAP will be required for a complete antitumor response in HCC.


Risk factors for tuberculosis in HIV-infected persons. A prospective cohort study. The Gruppo Italiano di Studio Tubercolosi e AIDS (GISTA).
To analyze how demographic, clinical, and laboratory characteristics influence the risk of tuberculosis in human immunodeficiency virus (HIV)-infected individuals; to examine the incidence of tuberculosis associated with change in skin test responsiveness in HIV-infected, tuberculin-negative, nonanergic individuals. Multicenter cohort study. Twenty-three infectious disease units in public hospitals in Italy. A consecutive sample of 3397 HIV-infected subjects were considered for entry in the study. Of these, 2695 who were followed up for at least 4 weeks were enrolled in the study; 739 subjects (27.4%) were unavailable for follow-up. The median duration of follow-up was 91 weeks. Culture-proven tuberculosis. Eighty-three episodes of tuberculosis were observed. Incidence rates of tuberculosis were 5.42 per 100 person-years among tuberculin-positive subjects, 3.00 per 100 person-years among anergic subjects, and 0.45 per 100 person-years among tuberculin-negative nonanergic subjects. In multivariate analysis, being tuberculin-positive (hazard ratio [HR], 9.94; 95% confidence interval [CI], 3.84 to 25.72) or anergic (HR, 3.35; 95% CI, 1.40 to 8.00), or having a CD4+ lymphocyte count less than 0.20 x 10(9)/L (HR, 4.87; 95% CI, 2.35 to 10.11) or between 0.20 and 0.35 x 10(9)/L (HR, 2.35; 95% CI, 1.09 to 5.05) were statistically significantly associated with the risk of tuberculosis. Incidence of tuberculosis increased with decreasing levels of CD4+ lymphocytes in the three groups of subjects with different skin test responsiveness. Skin tests were repeated 1 year after enrollment in 604 tuberculin-negative nonanergic subjects; three case of tuberculosis were observed among the 13 subjects who converted to tuberculin reactivity. Risk of tuberculosis in HIV-infected persons can be more precisely quantified by jointly considering skin test reactivity and CD4+ lymphocyte count. Periodic skin tests in tuberculin-negative nonanergic individuals can be useful in identifying individuals at high risk of active tuberculosis.


Xuemaitong granules attenuate carotid atherosclerosis by decreasing the expression of CD14+CD16+ monocytes, IL-6, TNF-α, and hsCRP.
Carotid atherosclerosis (CAS) has been extensively studied because its position can be easily observed. Our objective was to investigate the effects of Xuemaitong granules on the generation and activation of CD14+CD16+ monocytes on the inflammatory reaction in CAS patients. In this study, 22 male apolipoprotein E (apoE)-deficient mice were fed a high-fat diet for 13 weeks. After induction of an atherosclerotic plaque, the animals were randomly divided into the Xuemaitong granule group (450.5 mg/kg via intragastric administration, N=11) and the control group (equal volume saline via intragastric administration, N=11). Venous blood was obtained to analyze monocyte and CD14+CD16+ inflammatory monocyte levels, as well as interleukin (IL)-6, tumor necrosis factor (TNF)-a, and high-sensitivity C-reactive protein (hsCRP). For clinical studies, 100 CAS patients received oral administration of Xuemaitong granules for 6 months. Monocytes, CD14+CD16+ inflammatory monocytes, and the inflammatory cytokines IL-6, TNF-a, and hsCRP were analyzed. Compared with the control group, a remarkable decrease in the number of monocytes and CD14+CD16+ inflammatory monocytes as well as TNF-a, hsCRP, and IL-6 was noted in the Xuemaitong group. Compared with before treatment levels, the proportions of monocytes and their subsets of CD14+CD16+ inflammatory monocytes and the concentration of the inflammatory cytokines IL-6, TNF-a, and hsCRP significantly decreased. Xuemaitong granules played a significant role in the anti-inflammatory reactions. In addition, the granules attenuated the expression of the CD14+CD16+ inflammatory monocytes, resulting in the downregulation of the cytokines IL-6, TNF-a, and hsCRP.


Direct comparison of progenitor cells derived from adipose, muscle, and bone marrow from wild-type or craniosynostotic rabbits.
Reports have identified cells capable of osteogenic differentiation in bone marrow, muscle, and adipose tissues, but there are few direct comparisons of these different cell types. Also, few have investigated the potential connection between a tissue-specific abnormality and cells derived from seemingly unrelated tissues. In this article, the authors compare cells isolated from wild-type rabbits or rabbits with nonsyndromic craniosynostosis, defined as the premature fusion of one or more of the cranial sutures. Cells were derived from bone marrow, adipose, and muscle of 10-day-old wild-type rabbits (n = 17) or from age-matched rabbits with familial nonsyndromic craniosynostosis (n = 18). Cells were stimulated with bone morphogenetic protein-4 (BMP4), and alkaline phosphatase expression and cell proliferation were assessed. In wild-type rabbits, cells derived from muscle had more alkaline phosphatase activity than cells derived from either adipose or bone marrow. The cells derived from craniosynostotic rabbit bone marrow and muscle were significantly more osteogenic than those derived from wild-type rabbits. Adipose-derived cells demonstrated no significant differences. Although muscle-derived cells were most osteogenic in wild-type rabbits, bone marrow-derived cells were most osteogenic in craniosynostotic rabbits. These results suggest that cells from different tissues have different potentials for differentiation. Furthermore, cells derived from rabbits with craniosynostosis were different from cells from wild-type rabbits. Interestingly, cells derived from the craniosynostotic rabbits were not uniformly more responsive compared with wild-type cells, suggesting that specific tissue-derived cells may react differently in individuals with craniosynostosis.


Ultrafast nonlinear optical spectroscopy of a dual-band negative index metamaterial all-optical switching device.
We study the nonlinear optical response of a fishnet structure-metamaterial all-optical switching device that exhibits two near-infrared negative-index resonances. We study and compare the nonlinear optical response at both resonances and identify transient spectral features associated with the negative index resonance. We see a significantly stronger response at the longer wavelength resonance, but identical temporal dynamics at both resonances, providing insight into separately engineering the switching time and switching ratio of such a fishnet structure metamaterial all-optical switch. We also numerically reproduce the nonlinear behavior of our device using the Drude conductivity model and a finite integration technique over wide spectral and pump fluence ranges. Thereby, we show that beyond the linear properties of the device, the magnitude of the pump-probe response is completely described by only two material parameters. These results provide insight into engineering various aspects of the nonlinear response of fishnet structure metamaterials.


Deconvolution of fully overlapped reflections from crystals of foot-and-mouth disease virus O1 G67.
Foot-and-mouth disease virus O(1) G67 forms crystals that appear similar to those of the closely related viruses O(1)K and O(1)BFS, both of which belong to space group I23. Statistical disorder in the O(1) G67 crystals means, however, that the measured diffraction data possess higher symmetry consistent with point group 432. It is shown that this is due to intimate twinning, with mosaic blocks randomly distributed between the two orientations. This results in a twofold loss of information due to the exact superimposition of non-identical reflections from the two orientations. A simple procedure has been devised to deconvolute these overlapped reflections by applying constraints in both real and reciprocal space. This procedure works well, providing interpretable electron-density maps for this virus. Other applications are discussed.


Overweight duration in older adults and cancer risk: a study of cohorts in Europe and the United States.
Recent studies have shown that cancer risk related to overweight and obesity is mediated by time and might be better approximated by using life years lived with excess weight. In this study we aimed to assess the impact of overweight duration and intensity in older adults on the risk of developing different forms of cancer. Study participants from seven European and one US cohort study with two or more weight assessments during follow-up were included (n = 329,576). Trajectories of body mass index (BMI) across ages were estimated using a quadratic growth model; overweight duration (BMI ≥ 25) and cumulative weighted overweight years were calculated. In multivariate Cox models and random effects analyses, a longer duration of overweight was significantly associated with the incidence of obesity-related cancer [overall hazard ratio (HR) per 10-year increment: 1.36; 95 % CI 1.12-1.60], but also increased the risk of postmenopausal breast and colorectal cancer. Additionally accounting for the degree of overweight further increased the risk of obesity-related cancer. Risks associated with a longer overweight duration were higher in men than in women and were attenuated by smoking. For postmenopausal breast cancer, increased risks were confined to women who never used hormone therapy. Overall, 8.4 % of all obesity-related cancers could be attributed to overweight at any age. These findings provide further insights into the role of overweight duration in the etiology of cancer and indicate that weight control is relevant at all ages. This knowledge is vital for the development of effective and targeted cancer prevention strategies.


Simultaneous liver and kidney transplantation in elderly patients: Outcomes and validation of a clinical risk score for patient selection.
Introduction and aim. Many transplant programs have expanded eligibility to include patients previously ineligible because of advanced age. Outcomes of simultaneous liver-kidney transplantation (SLK) in recipients with advanced age are not known. Data from patients undergoing transplantation between 2002 and 2015 were obtained from the UNOS Standard Analysis and Research file. SLK recipients aged ≥ 65 years (N = 677), SLK recipients aged < 65 years (N = 4517), and recipients of liver transplant alone(LTA) aged ≥ 65 years(N = 8495) were compared. Recipient characteristics were similar between the SLK groups. Similar patient and graft survival were observed in SLK recipients aged ≥ 65 years compared to SLK recipients aged < 65 years and LTA recipients aged ≥ 65 years. Importantly, in a subgroup analysis, superior survival was seen in the SLK group aged ≥ 65 years compared to LTA recipients aged ≥ 65 years who underwent dialysis in the week prior to transplantation (p < 0.001). A prediction model of patient survival was developed for the SLK group aged ≥ 65 years with predictors including: age ≥ 70 years (3 points), calculated MELD score (-1 to 2 points), and recipient ventilator status at the time of SLK (4 points). The risk score predicted patient survival, with a significantly inferior survival seen in patients with a score ≥ 4 (p < 0.001). Age should not be used as a contraindication for SLK transplantation. The validated scoring system provides a guide for patient selection and can be used when evaluating elderly patients for SLK transplantation listing.


From signal transduction to signal interpretation: an alternative model for the molecular function of insulin receptor substrates.
The insulin receptor (IR) recruits adaptor proteins, so-called insulin receptor substrates (IRS), to connect with downstream signalling pathways. A family of IRS proteins was defined based on three major common structural elements: Amino-terminal PH and PTB domains that mediate protein-lipid or protein-protein interactions, mostly carboxy-terminal multiple tyrosine residues that serve as binding sites for proteins that contain one or more SH2 domains and serine/threonine-rich regions which may be recognized by negative regulators of insulin action. The current model for the role of IRS proteins therefore combines an adaptor function with the integration of mostly negative input from other signal transduction cascades allowing for modulation of signalling amplitude. In this review we propose an extended version of the adaptor model that can explain how signalling specificity could be implemented at the level of IRS proteins.


COVID-19's natural course among ambulatory monitored outpatients.
Research objective was to detail COVID-19's natural trajectory in relation to the Czech population's viral load. Our prospective detailed daily questionnaire-based telemonitoring study evaluated COVID-19's impact among 105 outpatients. In accordance with government quarantine requirements, outpatients were divided into a cohort with two negative tests at the end of the disease (40 patients) and a cohort with a new algorithm (65 patients) following a 14-day quarantine. Median follow-up differed significantly between the 2 groups (23 days vs. 16 days). Only 6% of patients were asymptomatic during the entire telemonitoring period. Another 13% of patients were diagnosed asymptomatic, as suspected contacts, yet later developed symptoms, while the remaining 81% were diagnosed as symptomatic on average 6 days following symptom onset. Telemonitoring enabled precise symptom status chronicling. The most frequently reported complaints were fevers, respiratory issues, and anosmia. Six patients were eventually hospitalized for complications detected early after routine telemonitoring. During the extended follow-up (median 181 days), anosmia persisted in 26% of patients. 79% of patients in the new quarantine algorithm cohort reported no symptoms on day 11 compared to just 56% of patients in the two negative test cohort upon first testing negative (median-19 days). The highest viral load occurred within 0-2 days of initial symptom onset. Both the PCR viral load and two consecutive PCR negative sample realizations indicated high interindividual variability with a surprisingly fluctuating pattern among 43% of patients. No definitive COVID-19 symptoms or set of symptoms excepting anosmia (59%) and/or ageusia (47%) were identified. No preexisting medical conditions specifically foreshadowed disease trajectory in a given patient. Without a PCR negativity requirement for quarantine cessation, patients could exhibit fewer symptoms. Our study therefore highlights the urgent need for routine ambulatory patient telemedicine monitoring, early complication detection, intensive mass education connecting disease demeanor with subsequent swift diagnostics, and, notably, the need to reevaluate and modify quarantine regulations for better control of SARS-CoV-2 proliferation.


The fat-1 transgene in mice increases antioxidant potential, reduces pro-inflammatory cytokine levels, and enhances PPAR-gamma and SIRT-1 expression on a calorie restricted diet.
Both n-3 fatty acids (FA) and calorie-restriction (CR) are known to exert anti-inflammatory and anti-oxidative effects in animals and humans. In this study, we investigated the synergistic anti-inflammatory and anti-oxidative capacity of n-3 FA and CR using Fat-1 transgenic mice (Fat-1) that are capable of converting n-6 FA to n-3 FA endogenously. Wild type (WT) and Fat-1 mice were maintained on ad libitum (AL) or CR (40% less than AL) AIN-93 diet supplemented with 10% corn oil (rich in n-6 FA) for 5 months. Significantly lower levels of n-6/n-3 FA ratio were observed in serum, muscle and liver of Fat-1 mice fed AL or CR as compared to that of WT mice fed AL or CR. Muscle catalase (CAT), super oxide dismutase (SOD), glutathione peroxidase (GPX) activities, and liver CAT and SOD activities were found higher in Fat-1 mice as compared to that of WT mice. These activities were more pronounced in Fat-1/CR group as compared to other groups. Serum pro-inflammatory markers, such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 were found lower in Fat-1 mice, as compared to that of WT mice. This anti-inflammatory effect was also more pronounced in Fat-1/CR group as compared to that of other groups. Furthermore, significantly higher levels of peroxisome proliferator-activated receptor (PPAR)-gamma and life prolonging gene, sirtuin (SIRT)-1 expression were found in liver of Fat-1/CR mice, as compared to that of WT/CR mice. These data suggest that n-3 FA along with moderate CR may prolong lifespan by attenuating inflammation and oxidative stress.


Human JCV infections as a bio-anthropological marker of the formation of Brazilian Amazonian populations.
JC polyomavirus (JCV) is a member of the Polyomaviridae family. It presents a tropism to kidney cells, and the infection occurs in a variety of human population groups of different ethnic background. The present study investigated the prevalence of JCV infection among human populations from the Brazilian Amazon region, and describes the molecular and phylogenetic features of the virus. Urine samples from two urban groups of Belém (healthy subjects), one Brazilian Afro-descendant "quilombo" from the Rio Trombetas region, and native Indians from the Wai-Wai, Urubu-Kaapor, Tembé, Assurini, Arara do Laranjal, Aukre, Parakanã, Surui and Munduruku villages were investigated for the presence of the virus by amplifying VP1 (230 bp) and IG (610 bp) regions using a polymerase chain reaction. Nucleotide sequences (440 nucleotides, nt) from 48 samples were submitted to phylogenetic analysis. The results confirmed the occurrence of types A (subtype EU), B (subtypes Af-2, African and MY, Asiatic) and C (subtype Af-1) among healthy subjects; type B, subtypes Af-2 and MY, among the Afro-Brazilians; and type B, subtype MY, within the Surui Indians. An unexpected result was the detection of another polyomavirus, the BKV, among Afro-descendants. The present study shows, for the first time, the occurrence of JC and BK polyomaviruses infecting humans from the Brazilian Amazon region. The results show a large genetic variability of strains circulating in the region, infecting a large group of individuals. The presence of European, Asiatic and African subtypes associated to the ethnic origin of the population samples investigated herein, highlights the idea that JCV is a fairly good marker for studying the early migration of human populations, reflecting their early and late history. Furthermore, the identification of the specific mutations associated to the virus subtypes, suggests that these mutations have occurred after the entrance of the virus in the Amazon region of Brazil.


A dominant-negative mutant inhibits multiple prion variants through a common mechanism.
Prions adopt alternative, self-replicating protein conformations and thereby determine novel phenotypes that are often irreversible. Nevertheless, dominant-negative prion mutants can revert phenotypes associated with some conformations. These observations suggest that, while intervention is possible, distinct inhibitors must be developed to overcome the conformational plasticity of prions. To understand the basis of this specificity, we determined the impact of the G58D mutant of the Sup35 prion on three of its conformational variants, which form amyloids in S. cerevisiae. G58D had been previously proposed to have unique effects on these variants, but our studies suggest a common mechanism. All variants, including those reported to be resistant, are inhibited by G58D but at distinct doses. G58D lowers the kinetic stability of the associated amyloid, enhancing its fragmentation by molecular chaperones, promoting Sup35 resolubilization, and leading to amyloid clearance particularly in daughter cells. Reducing the availability or activity of the chaperone Hsp104, even transiently, reverses curing. Thus, the specificity of inhibition is determined by the sensitivity of variants to the mutant dosage rather than mode of action, challenging the view that a unique inhibitor must be developed to combat each variant.


Accuracy of diagnosis and health service codes in identifying frailty in Medicare data.
Capturing frailty within administrative claims data may help to identify high-risk patients and inform population health management strategies. Although it is common to ascertain frailty status utilizing claims-based surrogates (e.g. diagnosis and health service codes) selected according to clinical knowledge, the accuracy of this approach has not yet been examined. We evaluated the accuracy of claims-based surrogates against two clinical definitions of frailty. This cross-sectional study was conducted in a Health and Retirement Study subsample of 3097 participants, aged 65 years or older and with at least 12-months of continuous fee-for-service Medicare enrollment. We defined 18 previously utilized claims-based surrogates of frailty from Medicare data and evaluated each against clinical reference standards, ascertained from a direct examination: a deficit accumulation frailty index (FI) (range: 0-1) and frailty phenotype. We also compared the accuracy of the total count of 18 claims-based surrogates with that of a validated claims-based FI model, comprised of 93 claims-based variables. 19% of participants met clinical criteria for the clinical frailty phenotype. The mean clinical FI for our sample was 0.20 (standard deviation 0.13). Hospital Beds and associated supplies was the claims-based surrogate associated with the highest clinical FI (mean FI 0.49). Claims-based surrogates had low sensitivity ranging from 0.01 (cachexia, adult failure to thrive, anorexia) to 0.38 (malaise and fatigue) and high specificity ranging from 0.79 (malaise and fatigue) to 0.99 (cachexia, adult failure to thrive, anorexia) in discriminating the clinical frailty phenotype. Compared with a validated claims-based FI, the total count of claims-based surrogates demonstrated lower Spearman correlation with the clinical FI (0.41 [95% CI 0.38-0.44] versus 0.59 [95% CI, 0.56-0.61]) and poorer discrimination of the frailty phenotype (C-statistics 0.68 [95% CI, 0.66-0.70] versus 0.75 [95% CI, 0.73-0.77]). Claims-based surrogates, selected according to clinical knowledge, do not accurately capture frailty in Medicare claims data. A simple count of claims-based surrogates improves accuracy but remains inferior to a claims-based FI model.


Comparative efficacy of nonpharmacological interventions on sleep quality in people who are critically ill: A systematic review and network meta-analysis.
Disrupted sleep is a critical and highly prevalent concern among critically ill patients requiring intensive care. However, the question of which nonpharmacological intervention represents the best strategy for improving sleep quality remains unanswered. To compare the efficacy of nonpharmacological interventions in improving sleep quality in people who are critically ill. Databases, namely PubMed, Embase, CINAHL, and ProQuest Dissertations and Theses A&I, were searched from their inception up until January 15, 2021, for relevant randomised controlled trials. No language or time period restrictions were applied. Only randomised controlled trials examining the effects of nonpharmacological interventions on sleep among adults (aged ≥18 years) admitted to an intensive care unit were included. A random-effects model was used for data analyses. The study protocol was registered at PROSPERO (CRD42021232004). Twenty randomised controlled trials involving 1,207 participants were included. Music combined with earplugs and eye masks (standardised mean difference =1.64), eye masks alone (0.98), aromatherapy (0.87), and earplugs combined with eye masks (0.61) significantly improved sleep quality compared with routine care (all p <0.05). Music combined with earplugs and eye masks significantly enhanced sleep quality in comparison with music (1.34), earplugs combined with eye masks (1.03), and nursing intervention (1.76, all p <0.05). Earplugs alone was less likely to have effects on sleep quality improvement compared with routine care. Eye masks alone and music combined with earplugs and eye masks appear to be the most effective interventions for improving sleep quality in people who are critically ill. Critical care nurses should incorporate the use of eye masks alone or music combined with eye masks into sleep care.


Diphenylpyrazoles as replication protein a inhibitors.
Replication Protein A is the primary eukaryotic ssDNA binding protein that has a central role in initiating the cellular response to DNA damage. RPA recruits multiple proteins to sites of DNA damage via the N-terminal domain of the 70 kDa subunit (RPA70N). Here we describe the optimization of a diphenylpyrazole carboxylic acid series of inhibitors of these RPA-protein interactions. We evaluated substituents on the aromatic rings as well as the type and geometry of the linkers used to combine fragments, ultimately leading to submicromolar inhibitors of RPA70N protein-protein interactions.


Risk of End-Stage Renal Disease in Patients With Systemic Lupus Erythematosus and Diabetes Mellitus: A Danish Nationwide Cohort Study.
The risk of end-stage renal disease (ESRD) is increased in patients with systemic lupus erythematosus (SLE). This study was undertaken to determine whether diabetes mellitus (DM) increases ESRD risk in a large inception cohort of SLE patients. By means of the Danish National Patient Registry, we identified 3,178 adult patients diagnosed as having SLE between January 1, 1996, and July 31, 2018. DM was defined as the date of first hospital contact for DM or date of a first prescription of an antidiabetic drug. ESRD was defined as first registration of dialysis, renal transplant, or terminal renal insufficiency in the Danish National Patient Registry. ESRD incidence was compared between SLE patients with DM (SLE-DM) and those without DM (SLE-non-DM). Hazard ratios (HRs), adjusted for sex, age, educational level, and occupational status at baseline were calculated for sex, age, educational level, and hypertension (at baseline or during follow-up) strata. The overall hazard ratio (HR) was also adjusted for hypertension. The SLE-DM group included 290 patients, of whom 77% were female, compared with 85% of the 2,859 patients in the SLE-non-DM group. SLE-DM patients had a 3 times higher risk of ESRD compared with SLE-non-DM patients (multivariable-adjusted HR 3.3 [95% confidence interval 1.8-6.1]). In stratified multivariable-adjusted analyses, DM increased the rate of ESRD in women and men, patients ≥50 years old at baseline, those with low educational level at baseline, and those with concomitant hypertension. Our findings indicate that SLE patients with DM have a markedly higher risk of developing ESRD compared with SLE patients without DM.


Prevalence of Metabolic Syndrome in Patients with Schizophrenia in Korea: A Multicenter Nationwide Cross-Sectional Study.
We designed a nationwide study with limited exclusion criteria to investigate the prevalence of metabolic syndrome (MetS) in Korea and its relationship with antipsychotic medications. This multicenter, cross-sectional, and observational study included patients diagnosed with schizophrenia or schizoaffective disorder. Sixteen hospitals enrolled 845 patients aged 18 to 65 years prescribed any antipsychotic medication between August 2011 and August 2013. MetS was diagnosed using the criteria of the modified Adult Treatment Panel III of the National Cholesterol Education Program with the Korean abdominal obesity definition (waist circumference ≥85 cm in women, ≥90 cm in men). The prevalence of MetS in all patients was 36.5% and was significantly higher in men than women (men, 40.8%; women, 32.2%) and was significantly correlated with age [odds ratio (OR) 1.02] and duration of illness (OR 1.03). The prevalence of MetS across antipsychotic drugs in the major monotherapy group was as follows: 18.8% for quetiapine, 22.0% for aripiprazole, 33.3% for both amisulpride and paliperidone, 34.0% for olanzapine, 35% for risperidone, 39.4% for haloperidol, and 44.7% for clozapine. The prevalence of MetS is very high in patients with schizophrenia or schizoaffective disorder. Screening and monitoring of MetS is also strongly recommended.


White matter changes linked to visual recovery after nerve decompression.
The relationship between the integrity of white matter tracts and cortical function in the human brain remains poorly understood. We investigate reversible white matter injury, in this case patients with compression of the optic chiasm by pituitary gland tumors, to study the structural and functional changes that attend spontaneous recovery of cortical function and visual abilities after surgical removal of the tumor and subsequent decompression of the nerves. We show that compression of the optic chiasm led to demyelination of the optic tracts, which reversed as quickly as 4 weeks after nerve decompression. Furthermore, variability across patients in the severity of demyelination in the optic tracts predicted visual ability and functional activity in early cortical visual areas. Preoperative measurements of myelination in the optic tracts predicted the magnitude of visual recovery after surgery. These data indicate that rapid regeneration of myelin in the human brain is a component of the normalization of cortical activity, and ultimately the recovery of sensory and cognitive function, after nerve decompression. More generally, our findings demonstrate the use of diffusion tensor imaging as an in vivo measure of myelination in the human brain.


A Community-based Oral Health Promotion Model for HIV Patients in Nairobi, East District in Kenya: a Study Protocol.
General HIV-related orofacial lesions, most commonly oropharyngeal candidiasis, have a typical clinical appearance and can be recognised by members of the community. Although affected patients often experience pain leading to compromised eating and swallowing, barriers such as social stigma and lack of knowledge regarding available services may prevent them from seeking early care. Educating the community about these lesions through community health workers (CHWs) who are democratically elected community members may encourage individuals affected to seek early oral healthcare in the health facilities. A health facility (HF) is a health centre mainly run by clinical officers (CO), i.e. personnel with a 3-year medical training, and nurses. This study aims to evaluate the effect of a CHW training programme on: i) their knowledge and recognition of HIV-related oral-facial lesions at a community level; and ii) referral of affected patients from the community to the HFs. All 800 CHWs in 2 administrative divisions of Nairobi East District (test group n=400; control group n=400) will be selected. The test group will receive training. CHWs in both groups will be assessed at 4 time points: -3, 0, +3 and +6 months with reference to the training on: i) their knowledge of HIV-related orofacial lesions (using a written questionnaire); and ii) their performance in referring affected patients to the HFs (using clinical data). Early recognition of HIV-related orofacial lesions at a community level will prompt community members to seek early oral care, leading to early HIV testing and counselling regarding failure of antiretroviral therapy, while treatment outcomes are still favourable.


Age-related decline in oligodendrogenesis retards white matter repair in mice.
Aging is one of the major risk factors for white matter injury in cerebrovascular disease. However, the effects of age on the mechanisms of injury/repair in white matter remain to be fully elucidated. Here, we ask whether, compared with young brains, white matter regions in older brains may be more vulnerable in part because of decreased rates of compensatory oligodendrogenesis after injury. A mouse model of prolonged cerebral hypoperfusion was prepared by bilateral common carotid artery stenosis in 2-month and 8-month-old mice. Matching in vitro studies were performed by subjecting oligodendrocyte precursor cells to sublethal 7-day CoCl2 treatment to induce chemical hypoxic stress. Baseline myelin density in the corpus callosum was similar in 2-month and 8-month-old mice. But after induction of prolonged cerebral hypoperfusion, older mice showed more severe white matter injury together with worse deficits in working memory. The numbers of newborn oligodendrocytes and their precursors were increased by cerebral hypoperfusion in young mice, whereas these endogenous responses were significantly dampened in older mice. Defects in cyclic AMP response element-binding protein signaling may be involved because activating cyclic AMP response element-binding protein with the type-III phosphodiesterase inhibitor cilostazol in older mice restored the differentiation of oligodendrocyte precursor cells, alleviated myelin loss, and improved cognitive dysfunction during cerebral hypoperfusion. Cell culture systems confirmed that cilostazol promoted the differentiation of oligodendrocyte precursor cells. An age-related decline in cyclic AMP response element-binding protein-mediated oligodendrogenesis may compromise endogenous white matter repair mechanisms, and therefore, drugs that activate cyclic AMP response element-binding protein signaling provide a potential therapeutic approach for treating white matter injury in aging brains.


Parent Artery Occlusion for Unruptured Cerebral Aneurysms: Results of the Japanese Registry of Neuroendovascular Therapy 3.
Although the current standard treatment for unruptured aneurysms comprises surgical clipping or endovascular coiling, these techniques are not suitable for some cases, such as large, giant, and fusiform aneurysms. Endovascular parent artery occlusion (PAO), which includes internal trapping and proximal occlusion, is a well-established alternative treatment for such cases. Here, we retrospectively reviewed PAO cases from the Japanese Registry of Neuroendovascular Therapy 3, a nation-wide survey of all neuroendovascular therapy cases between 2010 and 2014. This dataset included 274 procedures with a mean patient age of 57.1 years and 55.4% female patients. For the treatment strategy, internal trapping was selected in 213 aneurysm cases (77.7%) and proximal occlusion in 61 aneurysm cases (22.2%). Most of the procedures were successfully completed (272/274: 99.2%). Immediately after treatment, angiographical complete occlusion was achieved in 248 cases (90.5%). Although the feasibility of this technique was excellent, there were 60 periprocedural complications (21.8%), including 48 ischemic complications (17.5%), seven hemorrhagic complications (2.5%). Overall, morbidity and mortality at 30 days postoperative were 5.8% and 0.7%, respectively. Among the pretreatment variables, a patient age of 70 and older was associated with ischemic complications [odds ratio (OR); 2.34, 95% confidence interval (CI); 1.02-5.25; P = 0.04] and a small aneurysm size (<5 mm) was associated with hemorrhagic complications (OR; 9.85, 95% CI; 1.07-221.0; P = 0.04) by multivariate analysis. In conclusion, PAO for unruptured cerebral aneurysms is feasible, but is associated with a complication rate of approximately 20%. Various alternative treatment options should be carefully considered with deconstructive strategies.


A systematic review of therapeutic interventions to reduce acute and chronic post-surgical pain after amputation, thoracotomy or mastectomy.
Perioperative neuropathic pain is under-recognized and often undertreated. Chronic pain may develop after any routine surgery, but it can have a far greater incidence after amputation, thoracotomy or mastectomy. The peak noxious barrage due to the neural trauma associated with these operations may be reduced in the perioperative period with the potential to reduce the risk of chronic pain. A systematic review of the evidence for perioperative interventions reducing acute and chronic pain associated with amputation, mastectomy or thoracotomy. Thirty-two randomized controlled trials met the inclusion criteria. Gabapentinoids reduced pain after mastectomy, but a single dose was ineffective for thoracotomy patients who had an epidural. Gabapentinoids were ineffective for vascular amputees with pre-existing chronic pain. Venlafaxine was associated with less chronic pain after mastectomy. Intravenous and topical lidocaine and perioperative EMLA (eutectic mixture of local anaesthetic) cream reduced the incidence of chronic pain after mastectomy, whereas local anaesthetic infiltration appeared ineffective. The majority of the trials investigating regional analgesia found it to be beneficial for chronic symptoms. Ketamine and intercostal cryoanalgesia offered no reduction in chronic pain. Total intravenous anaesthesia (TIVA) reduced the incidence of post-thoracotomy pain in one study, whereas high-dose remifentanil exacerbated chronic pain in another. Appropriate dose regimes of gabapentinoids, antidepressants, local anaesthetics and regional anaesthesia may potentially reduce the severity of both acute and chronic pain for patients. Ketamine was not effective at reducing chronic pain. Intercostal cryoanalgesia was not effective and has the potential to increase the risk of chronic pain. TIVA may be beneficial but the effects of opioids are unclear.


Methyl donor supply to heat stress-challenged polymorphonuclear leukocytes from lactating Holstein cows enhances 1-carbon metabolism, immune response, and cytoprotective gene network abundance.
Mechanisms controlling immune function of dairy cows are dysregulated during heat stress (HS). Methyl donor supply-methionine (Met) and choline (Chol)-positively modulates innate immune function, particularly antioxidant systems of polymorphonuclear leukocytes (PMN). The objective of this study was to investigate the effect of Met and Chol supply in vitro on mRNA abundance of genes related to 1-carbon metabolism, inflammation, and immune function in short-term cultures of PMN isolated from mid-lactating Holstein cows in response to heat challenge. Blood PMN were isolated from 5 Holstein cows (153 ± 5 d postpartum, 34.63 ± 2.73 kg/d of milk production; mean ± SD). The PMN were incubated for 2 h at thermal-neutral (37°C; TN) or heat stress (42°C; HS) temperatures with 3 levels of Chol (0, 400, or 800 μg/mL) or 3 ratios of Lys:Met (Met; 3.6:1, 2.9:1, or 2.4:1). Supernatant concentrations of IL-1β, IL-6, and tumor necrosis factor-α were measured via bovine-specific ELISA. Fold-changes in mRNA abundance were calculated separately for Chol and Met treatments to obtain the fold-change response at 42°C (HS) relative to 37°C (TN). Data were subjected to ANOVA using PROC MIXED in SAS (SAS Institute Inc., Cary, NC). Orthogonal contrasts were used to determine the linear or quadratic effect of Met and Chol for mRNA fold-change and supernatant cytokine concentrations. Compared with PMN receiving 0 μg of Chol/mL, heat-stressed PMN supplemented with Chol at 400 or 800 μg/mL had greater fold-change in abundance of CBS, CSAD, GSS, GSR, and GPX1. Among genes associated with inflammation and immune function, fold-change in abundance of TLR2, TLR4, IRAK1, IL1B, and IL10 increased with 400 and 800 μg of Chol/mL compared with PMN receiving 0 μg of Chol/mL. Fold-change in abundance of SAHH decreased linearly at increasing levels of Met supply. A linear effect was detected for MPO, NFKB1, and SOD1 due to greater fold-change in abundance when Met was increased to reach Lys:Met ratios of 2.9:1 and 2.4:1. Although increasing Chol supply upregulated BAX, BCL2, and HSP70, increased Met supply only upregulated BAX. Under HS conditions, enhancing PMN supply of Chol to 400 μg/mL effectively increased fold-change in abundance of genes involved in antioxidant production (conferring cellular processes protection from free radicals and reactive oxygen species), inflammatory signaling, and innate immunity. Although similar outcomes were obtained with Met supply at Lys:Met ratios of 2.9:1 and 2.4:1, the response was less pronounced. Both Chol and Met supply enhanced the cytoprotective characteristics of PMN through upregulation of heat shock proteins. Overall, the modulatory effects detected in the present experiment highlight an opportunity to use Met and particularly Chol supplementation during thermal stress.


Newborn screening and the obstetrician.
Newborn screening is the largest genetic screening program in the United States with approximately four million newborns screened yearly. It has been available and in continuous development for more than 50 years. Each state manages, funds, and maintains its own individual program, which encompasses newborn screening as well as the diagnosis and coordination of care for affected infants and children. The ideal disorder for screening is one in which newborn intervention prevents later disabilities or death for infants who may appear normal at birth. There are 31 core conditions that are currently recommended for incorporation into state screening programs. To obtain a sample, several drops of blood are collected from the newborn's heel and applied to filter paper. Although testing for core disorders is fairly standardized, more extensive screening varies by state and the rigorous evaluation of new disorders for inclusion in state screening panels is ongoing. As genomic medicine becomes more accessible, screening newborns for chronic diseases that may affect their long-term health will need to be addressed as well as the use of the residual blood spots for research. Obstetric providers should, at some time during pregnancy, review the basic process of newborn screening with parents to prepare them for this testing in the neonatal period. This information can be reviewed as it best suits incorporation in an individual's practice; verbal discussion and the distribution of written materials with resources for further information are encouraged.


Behavioral changes with paranoia in an elderly woman taking atorvastatin.
There have been a number of published reports of central nervous system (CNS) adverse effects with statins. A 79-year-old woman developed paranoia, anxiety, and behavioral changes approximately 2.5 weeks after starting atorvastatin 10 mg/d. The patient had no other medication changes at this time. After 2 months of therapy, the patient discontinued atorvastatin, and her symptoms fully resolved after 4 days. This is the first case report, to our knowledge, describing paranoia as one of the symptoms associated with statin therapy. Our report suggests an adverse reaction due to the initiation of atorvastatin via the temporal relationship between the start of atorvastatin and symptom onset, as well as termination of therapy and subsequent symptom disappearance. Use of the Naranjo adverse drug reaction probability scale to assess causality revealed a "probable" association (score, 5) for this adverse event. This report emphasizes the possibility of paranoia as a CNS adverse effect due to statin therapy. Statins are frequently used in older populations and should therefore be considered when such CNS adverse effects occur during therapy.