Effects of 3-methylcholanthrene pretreatment on microsomal hydroxylation of 2-acetamidofluorene by various rat hepatomas.
1. The effects of 3-methylcholanthrene pretreatment on both N- and ring hydroxylation of 2-acetamidofluorene by microsomal preparations from various well-differentiated and poorly differentiated hepatomas, primary tumours and their host livers and kidneys were studied. 2. Well-differentiated Morris hepatomas 5123C, 5123D, 5123CTC and 7800 and their host livers had low hydroxylating activity. Pretreatment with 3-methylcholanthrene caused a several-fold increase in both N- and ring hydroxylation in the host livers whereas in all tumours except 5123CTC it caused a many-fold increase only in ring hydroxylation. 5123CTC tumour in addition showed a fourfold increase in N-hydroxylating activity. 3. Hydroxylating activities of poorly differentiated Morris hepatoma 7288CTC and Novikoff hepatoma were low and they could not be altered by 3-methylcholanthrene pretreatment. 4. Primary hepatomas produced by administration of 4-dimethylamino-3'-methylazobenzene could be stimulated to some extent on 3-methylcholanthrene pretreatment; however, primary mammary tumour produced by administration of 3-methylcholanthrene was not responsive to 3-methycholanthrene pretreatment. 5. Like host livers, kidneys of tumour-bearing animals could also be stimulated to some extent by 3-methylcholanthrene pretreatment.


A Rapid Tactile-Motor Reflex Automatically Guides Reaching toward Handheld Objects.
The ability to respond quickly and effectively when objects in the world suddenly change position is essential for skilled action, and previous work has documented how unexpected changes in the location of a visually presented target during reaching can elicit rapid reflexive (i.e., automatic) corrections of the hand's trajectory [1-12]. In object manipulation and tool use, the sense of touch can also provide information about changes in the location of reach targets. Consider the many tasks where we reach with one hand to part of an object grasped by the other hand: reaching to a berry while holding a branch, reaching for a cap while grasping a bottle, and reaching toward a dog's collar while holding the dog's leash. In such cases, changes in the position of the reach target, due to wind, slip, or an active agent, can be detected, in principle, through touch. Here, we show that when people reach with their right hand to a target attached to the far end of a rod contacted, at the near end, by their left hand, an unexpected change in target location caused by rod rotation rapidly evokes an effective reach correction. That is, spatial information about a change in target location provided by tactile inputs to one hand elicits a rapid correction of the other hand's trajectory. In addition to uncovering a tactile-motor reflex that can support manipulatory actions, our results demonstrate that automatic reach corrections to moving targets are not unique to visually registered changes in target location.


Differential display and cloning of messenger RNAs from human breast cancer versus mammary epithelial cells.
Identification of the genes that are specifically expressed in tumor cells but not in normal cells (oncogenes), or vice versa (tumor suppressor genes), is important for understanding the molecular basis of cancer. The differential display technique was applied to compare mRNAs from normal and tumor-derived human mammary epithelial cells, cultured under the same conditions. Complementary DNA fragments corresponding to several apparently differentially expressed mRNAs were recovered and sequenced. They exhibit characteristics of the 3' end of eukaryotic mRNA, as predicted by the method. A complementary DNA fragment seen only in the normal cell was used as a probe to isolate its corresponding complementary DNA clone from a library. Northern analysis confirmed its differential expression. Thus, this method can be used for detecting, cloning, and sequencing of genes that are unique to a host of biological and disease processes.


The dynamin-related GTPase Drp1 is required for embryonic and brain development in mice.
The dynamin-related guanosine triphosphatase Drp1 mediates the division of mitochondria and peroxisomes. To understand the in vivo function of Drp1, complete and tissue-specific mouse knockouts of Drp1 were generated. Drp1-null mice die by embryonic day 11.5. This embryonic lethality is not likely caused by gross energy deprivation, as Drp1-null cells showed normal intracellular adenosine triphosphate levels. In support of the role of Drp1 in organelle division, mitochondria formed extensive networks, and peroxisomes were elongated in Drp1-null embryonic fibroblasts. Brain-specific Drp1 ablation caused developmental defects of the cerebellum in which Purkinje cells contained few giant mitochondria instead of the many short tubular mitochondria observed in control cells. In addition, Drp1-null embryos failed to undergo developmentally regulated apoptosis during neural tube formation in vivo. However, Drp1-null embryonic fibroblasts have normal responses to apoptotic stimuli in vitro, suggesting that the apoptotic function of Drp1 depends on physiological cues. These findings clearly demonstrate the physiological importance of Drp1-mediated organelle division in mice.


Bile acids and bariatric surgery.
Bariatric surgery, specifically Roux-en-Y gastric bypass (RYGB) and vertical sleeve gastrectomy (VSG), are the most effective and durable treatments for morbid obesity and potentially a viable treatment for type 2 diabetes (T2D). The resolution rate of T2D following these procedures is between 40 and 80% and far surpasses that achieved by medical management alone. The molecular basis for this improvement is not entirely understood, but has been attributed in part to the altered enterohepatic circulation of bile acids. In this review we highlight how bile acids potentially contribute to improved lipid and glucose homeostasis, insulin sensitivity and energy expenditure after these procedures. The impact of altered bile acid levels in enterohepatic circulation is also associated with changes in gut microflora, which may further contribute to some of these beneficial effects. We highlight the beneficial effects of experimental surgical procedures in rodents that alter bile secretory flow without gastric restriction or altering nutrient flow. This information suggests a role for bile acids beyond dietary fat emulsification in altering whole body glucose and lipid metabolism strongly, and also suggests emerging roles for the activation of the bile acid receptors farnesoid x receptor (FXR) and G-protein coupled bile acid receptor (TGR5) in these improvements. The limitations of rodent studies and the current state of our understanding is reviewed and the potential effects of bile acids mediating the short- and long-term metabolic improvements after bariatric surgery is critically examined.


Time related changes in luteal prostaglandin synthesis and steroidogenic capacity during pregnancy, normal and antiprogestin induced luteolysis in the bitch.
In nonpregnant and pregnant dogs the corpora lutea (CL) are the only source of progesterone (P4) which shows an almost identical secretion pattern until the rapid decrease of P4 prior to parturition. For the nonpregnant dog clear evidence has been obtained that physiological luteal regression is devoid of a functional role of the PGF2alpha-system and seems to depend on the provision of StAR. Yet in pregnant dogs the rapid prepartal luteal regression, coinciding with an increase of PGF2alpha, may be indicative for different regulatory mechanisms. To assess this situation and by applying semi-quantitative Real Time (Taq Man) RT-PCR, expression patterns were determined for the following factors in CL of pregnant and prepartal dogs and of mid-pregnant dogs treated with the antiprogestin Aglepristone: cyclooxygenase 2 (Cox2), prostaglandin E2 synthase (PGES), prostaglandin F2alpha synthase (PGFS), its receptors (EP2, EP4 an FP), the steroidogenic acute regulatory protein (StAR), 3beta-hydroxysteroid-dehydrogenase (3betaHSD) and the progesterone receptor (PR). Peripheral plasma P4 concentrations were determined by RIA. CL were collected via ovariohysterectomy from pregnant bitches (n=3-5) on days 8-12 (Group 1, pre-implantation period), days 18-25 (Group 2, post-implantation period), days 35-40 (Group 3, mid-gestation period) and during the prepartal progesterone decline (Group 4). Additionally, CL were obtained from groups of 5 mid-pregnant dogs (days 40-45) 24h, respectively 72h after the second treatment with Aglepristone. Expression of Cox2 and PGES was highest during the pre-implantation period, that of PGFS and FP during the post-implantation period. EP4 and EP2 revealed a constant expression pattern throughout pregnancy with a prepartal upregulation of EP2. 3betaHSD and StAR decreased significantly from the pre-implatation period to prepartal luteolysis, it was matched by the course of P4 concentrations. Expression of the PR was higher during mid-gestation and prepartal luteolysis than in the two preceding periods. After application of Aglepristone the overall mRNA-expression resembled the situation during prepartal luteolysis except for EP2, which remained unchanged. These data suggest that - as in the nonpregnant bitch - also in the pregnant bitch luteal production of prostaglandins is associated with luteal support rather than luteolysis. On the other hand induction of luteolysis by the PR blocker Aglepristone points to a role of luteal P4 as an autocrine factor in a positive loop feedback system controlling the availability of P4, StAR and 3betaHSD.


Targeting the 5'-AMP-activated protein kinase and related metabolic pathways for the treatment of prostate cancer.
Increasing evidence suggests that prostate cancer cells undergo unique metabolic reprogramming during transformation. A master regulator of cellular homeostasis, 5'-AMP-activated protein kinase (AMPK), directs metabolic adaptation that supports the growth demands of rapidly dividing cancer cells. The utilization of AMPK as a therapeutic target may therefore provide an effective strategy in the treatment of prostate cancer. Our review describes the regulation of AMPK by androgens and upstream kinases including the calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) in prostate cancer. Oncogenic, AMPK-regulated pathways that direct various metabolic processes are also addressed. Furthermore, we discuss the role of AMPK in growth arrest and autophagy as a potential survival pathway for cancer cells. In addition, by regulating non-metabolic pathways, AMPK may stimulate migration and mitosis. Finally, this review summarizes efforts to treat prostate cancer with pharmacological agents capable of modulating AMPK signaling. Current research is primarily focused on developing drugs that activate AMPK as a treatment for prostate cancer. However, oncogenic aspects of AMPK signaling calls for caution about employing such therapies. We think that inhibitors of CaMKK2 or AMPK, or perhaps the modulation of downstream targets of AMPK, will gain importance in the clinical management of prostate cancer.


Salmonella dublin infection of a prosthetic vascular graft successfully treated with ciprofloxacin.
The treatment of an infected vascular graft invariably requires an extra-anatomic bypass through a non-infected tissue plane, as simple drainage with antibiotics is rarely successful. We report a case in which an established infection of a prosthetic vascular graft with Salmonella dublin was successfully treated with simple drainage and a prolonged course of oral ciprofloxacin.


Reproducibility of Objective and Subjective Markers of Exercise Recovery in College Aged Males.
This study investigated the reproducibility of objective and subjective parameters of recovery pre- and post-exercise in college-aged male athletes. Thirteen male (aged 19-22y) team sport players were assessed for a range of recovery markers before and 24 hours after a repeated sprint protocol. An identical procedure was followed one week later. Participants undertook two objective tests: creatine kinase (CK) and countermovement jump (CMJ) height; and two subjective tests: visual analogue scale (VAS) for muscle soreness and 5-item well-being questionnaire (WB). Coefficients of variation (CV) of CMJ and WB were lowest of all markers studied both pre (4.0% and 5.9%) and 24 hours post (7.7% and 7.1%) exercise, respectively. The CV of a single CMJ showed the highest reproducibility pre-exercise (4.0%) compared to taking the best or average of 2 or 3 jumps. Both CK and VAS had a high CV at pre (25.6% and 49.2%) and 24 hours post (44.5% and 44.8%) exercise. Moreover, while there was no difference between the change in CMJ, WB and VAS in response to exercise between weeks, the increase in CK was greater after the first compared to second exercise bout (mean 199.6 U/L vs 10.6 U/L change, p = 0.001), indicating a repeated bout effect. CK and VAS demonstrated poor reproducibility. However, single CMJ height and the WB questionnaire demonstrated a high reproducibility pre- and post-exercise and represent simple time-efficient objective and subjective methods to monitor recovery in this population.


Vitamin D and Osteoporosis in HIV/HCV Coinfected Patients: A Literature Review.
Vitamin D deficiency further increases the risk of osteoporosis in HIV-positive patients coinfected with hepatitis C virus (HCV); however, it is still unclear whether HCV-related increased fracture risk is a function of the severity of liver disease. The aim of this review was to identify studies on associative vitamin D deficiency patterns in high-risk populations such as HIV/HCV coinfected patients. We did this by searching MEDLINE and EMBASE databases, from inception to August 2014, and included bibliographies. The final 12 articles selected are homogeneous in terms of age but heterogeneous in terms of sample size, participant recruitment, and data source. Most of the HIV/HCV coinfected patients have less than adequate levels of vitamin D. After reviewing the selected articles, we concluded that vitamin D deficiency should be regarded as a continuum and that the lower limit of the ideal range is debatable. We found that vitamin D deficiency might influence liver disease progression in HIV/HCV coinfected patients. Methodological issues in evaluating vitamin D supplementation as a relatively inexpensive therapeutic option are discussed, as well as the need for future research, above all on its role in reducing the risk of HCV-related fracture by modifying liver fibrosis progression.


Social safety, self-rated general health and physical activity: changes in area crime, area safety feelings and the role of social cohesion.
The aim of this study was to examine whether changes over time in reported area crime and perceived area safety were related to self-rated general health and physical activity (PA), in order to provide support for a causal relationship between social safety and health. Additionally, we investigated whether social cohesion protects the residents against the negative impact of unsafe areas on health and PA. Multilevel logistic regression analyses were performed on Dutch survey data, including 47,926 respondents living in 2974 areas. An increase in area level unsafety feelings between 2009 and 2011 was associated with more people reporting poor general health in 2012 in that area, but was not related to PA. Changes in reported area crime were not related to either poor general health or PA. The social cohesion in the area did not modify the effect of changes in social safety on health and PA. The results suggest that tackling feelings of unsafety in an area might contribute to the better general health of the residents. Because changes in area social safety were not associated with PA, we found no leads that such health benefits were achieved through an increase in physical activity.


Cultured megakaryocytes: changes in the cytoskeleton after ADP-induced spreading.
Megakaryocytes from guinea pig bone marrow were isolated and maintained in liquid culture and were treated with ADP, thrombin, arachidonic acid, or collagen. Megakaryocytes spread with an active ruffled membrane in response to ADP (1-100 microM), thrombin (1.0 U/ml), and arachidonic acid (50 microM) but responded to collagen surfaces only if fibronectin was added to the cultures. Spreading could be blocked completely by dibutyryl cyclic AMP (dibutyryl cAMP) or isobutylmethylxanthine at 1 mM, as well as by cytochalasin D (2 microgram/ml), but not by colchicine up to 1 mg/ml. The distribution of contractile proteins was examined by immunofluorescence. In untreated, spherical cells, staining with antimyosin, antifilamin, anti-alpha-actinin, or with fluorescein-labeled subfragment 1 (FITC-S1) was diffuse and unpatterned. With antitubulin antibody, however, microtubules were seen in a dense array throughout the unspread cells. In actively ruffling spreading cells, myosin, filamin, and actin were visualized in the region of the ruffled membrane while alpha-actinin was seen most prominently in a band located proximal to the inner part of the ruffle. In fully spread cells, actin, myosin, filamin, and alpha-actinin were seen in filaments that filled the cytoplasm. Antimyosin and anti-alpha-actinin staining of the filaments was periodic with approximately 1 micrometer center-to-center spacing. Actin, filamin, and alpha-actinin were also identified in punctate spots throughout the spread cytoplasm. Microtubules were absent from the ruffle but filled the cytoplasm of fully spread cells. Rings, 1.5-2.5 micrometer in diameter, were seen with antitubulin in 13% of the spread cells. Our results show that megakaryocytes respond to platelet agonists, but typically by spreading, rather than extending, filopodia. From the changes in localization of contractile proteins and from time-lapse cinematography, we propose a model for cell spreading.


PRRT2 mutations in Japanese patients with benign infantile epilepsy and paroxysmal kinesigenic dyskinesia.
This study was performed to clarify the clinical features of Japanese patients with PRRT2 mutations. The PRRT2 gene was analyzed in 135 patients with benign infantile epilepsy (BIE) or paroxysmal kinesigenic dyskinesia (PKD) using a direct sequencing method: 92 patients had BIE alone, 25 had both BIE and PKD, and 18 had PKD alone. Of the cases, 105 were familial, and 30 were sporadic. Clinical information was collected using a structured questionnaire. PRRT2 mutations were identified in 104 patients. Among the familial cases, PRRT2 mutations were found in at least one individual in 21 of 28 families with BIE alone, in 26 of 27 families with infantile convulsions and choreoathetosis, and in 2 of 3 families with PKD alone. Among the sporadic cases, PRRT2 mutations were observed in 7 of 25 patients with BIE alone, in 1 of 1 patient with BIE and PKD, and in 3 of 4 patients with PKD alone. The c.649dupC mutation was the most frequent, followed by the c.981C > G mutation. Among the patients with epilepsy, the median age at BIE onset was 5 months, the median age at the last seizure was 6 months, and the median number of seizures was 5. PRRT2 mutations were found in 68% of Japanese probands with BIE or PKD. The phenotypes of BIE associated with PRRT2 mutations were consistent with those of BIE diagnosed clinically.


Effect of Intravenous Phenobarbital on Left Ventricular Myocardial Contractility Determined by Echocardiography in Children.
Animal studies and rare human studies have suggested a negative effect of barbiturates on cardiac function. Although intravenous (IV) phenobarbital is used routinely in children in the clinical setting, studies in children are lacking. We performed a study to evaluate effect of IV phenobarbital loading on myocardial systolic function of children. In a prospective pilot study in children without congenital heart defects, the effect of IV phenobarbital was evaluated on the left ventricular systolic function measured by ejection fraction (EF) by Simpson's method via an echocardiogram. Any child less than 18 years of age who received IV loading dose of at least 20 mg/kg of phenobarbital given as an infusion over 20 - 30 minutes for various medical indications was eligible to take part in the study. Three measurements of EF by an echocardiogram were made: before loading dose, 30 minutes after completion of the loading dose, and prior to the first maintenance dose. Relevant clinical data were recorded, including vital signs, immediately prior to each echocardiogram. Change of function as measured by EF over time was analyzed using linear mixed modeling methods. For this study, significant change in blood pressure was defined as a drop of at least 20 mmHg in systolic blood pressure. Ten children (70% female, age range two days to 8.2 years) were enrolled. Three had hypotension with a drop of systolic blood pressure greater than 20 mmHg from baseline. On examining the trajectory of EF on each individual graphically, the left ventricular EF tended to fall immediately following phenobarbital therapy and return to baseline on re-evaluation. These trajectories were statistically significant for EF. Phenobarbital had a direct and transient depressant effect on systolic function of the myocardium in one third of the cases. The depression in EF appeared to be transient with return to baseline in less than 24 hours. We recommend close monitoring with anticipation of decreased function in children when using IV phenobarbital.


Do the 2010 ACR/EULAR or ACR 1987 classification criteria predict erosive disease in early arthritis?
The new 2010 American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) classification criteria for rheumatoid arthritis (RA) aim at earlier diagnosis of RA compared to the 1987 ACR criteria. To evaluate the ability of the 2010 ACR/EULAR and the 1987 ACR classification criteria to predict radiographic progression after 10 years of follow-up. All early arthritis patients referred to Central Hospital in Jyväskylä from 1997 to 1999 (cases with peripheral joint synovitis, other specific diseases excluded) were included in this 10-year follow-up study. Radiographs of hands and feet were analysed according to Larsen on a scale of 0-100. At 10 years, 58% of the patients had an erosive disease (defined as Larsen ≥2 in at least one joint). The discriminative power of the 2010 ACR/EULAR and the 1987 ACR criteria (erosive disease at 10 years) were comparable, with area under the curve 0.72 (95% CI 0.65 to 0.79) (2010 ACR/EULAR criteria) and 0.65 (95% CI 0.58 to 0.72) (1987 ACR criteria). The respective sensitivities and specificities were 0.87 and 0.70, and 0.44 and 0.47. At 10 years, median (IQR) Larsen score was 6 (0, 15) among patients who had fulfilled both sets of criteria, 2 (0, 8) in those who met the 2010 ACR/EULAR and did not meet the ACR 1987 criteria, 0 (0, 5) in those who met ACR 1987 criteria but did not meet 2010 ACR/EULAR criteria, and 0 (0, 2) among patients who did not fulfil either of the criteria. The percentage of patients with erosions was 69%, 64%, 32% and 26%, respectively. The ability of the 2010 ACR/EULAR and 1987 ACR classification criteria to identify erosive disease in early arthritis is low. The discriminative power of the 2010 ACR/EULAR criteria of erosiveness in 10 years is slightly better than that of the 1987 ACR criteria.


Design, synthesis and bioactivity investigation of tetrandrine derivatives as potential anti-cancer agents.
Twenty-four 14-sulfonamide-tetrandrine derivatives as potential anti-cancer agents were synthesized. The synthetic derivatives were investigated for their cytotoxic activity against human cancer cell lines MDA-MB-231, PC3, WM9, HEL and K562. Initially, the IC50 values (50% inhibitory concentrations) of all of the compounds were determined. These derivatives exhibited potent, but distinct, inhibitory effects on the above-mentioned cell lines. Among them, compound 23, which was modified with a 2-naphthalenesulfonyl group at the 14-amino position, showed impressive inhibition of all five cancer cell lines, and especially of MDA-MB-231 cells with an IC50 value of 1.18 ± 0.14 μM. Further mechanism exploration showed that 23 induced potent apoptotic cell death on MDA-MB-231 cancer cells in a concentration-dependent manner. The results revealed that 23 might be a potential anti-cancer drug candidate.


Towards evidence-based and inclusive models of peer support for long covid: A hermeneutic systematic review.
Since the first wave of COVID-19 in March 2020 the number of people living with post-COVID syndrome has risen rapidly at global pace, however, questions still remain as to whether there is a hidden cohort of sufferers not accessing mainstream clinics. This group are likely to be constituted by already marginalised people at the sharp end of existing health inequalities and not accessing formal clinics. The challenge of supporting such patients includes the question of how best to organise and facilitate different forms of support. As such, we aim to examine whether peer support is a potential option for hidden or hardly reached populations of long COVID sufferers with a specific focus on the UK, though not exclusively. Through a systematic hermeneutic literature review of peer support in other conditions (57 papers), we evaluate the global potential of peer support for the ongoing needs of people living with long COVID. Through our analysis, we highlight three key peer support perspectives in healthcare reflecting particular theoretical perspectives, goals, and understandings of what is 'good health', we call these: biomedical (disease control/management), relational (intersubjective mutual support) and socio-political (advocacy, campaigning & social context). Additionally, we identify three broad models for delivering peer support: service-led, community-based and social media. Attention to power relations, social and cultural capital, and a co-design approach are key when developing peer support services for disadvantaged and underserved groups. Models from other long-term conditions suggest that peer support for long COVID can and should go beyond biomedical goals and harness the power of relational support and collective advocacy. This may be particularly important when seeking to reduce health inequalities and improve access for a potentially hidden cohort of sufferers.


Peripheral defocus and myopia progression in myopic children randomly assigned to wear single vision and progressive addition lenses.
To determine the effect of progressive addition lenses (PALs) and single vision lenses (SVLs) on peripheral defocus in myopic children, and to compare the effect of myopic versus hyperopic peripheral defocus on foveal myopia progression. Eighty-four myopic children aged 6 to 11 years with spherical equivalent (SE) cycloplegic autorefraction between -0.75 diopters (D) and -4.50 D were randomly assigned to wear SVLs or PALs. Aberrometry measurements of the eye and spectacles were made centrally, 30° nasally, temporally, and superiorly, and 20° inferiorly on the retina using a Complete Ophthalmic Analysis System for Vision Research (COAS-VR). The association between peripheral defocus and the 1-year change in central myopia was investigated. SVLs caused a hyperopic shift in peripheral defocus at all locations (all P ≤ 0.0003). PALs caused a myopic shift in peripheral defocus in three of four locations measured (all P ≤ 0.01) with the greatest shift superiorly due to the PAL addition (-1.04 ± 0.30 D). Superior retinal defocus when wearing either SVLs or PALs was associated with the 1-year change in central myopia. The adjusted 1-year change in central SE myopia was -0.38 D for children with absolute superior myopic defocus (n = 67) and -0.65 D for children with absolute superior hyperopic defocus (n = 17; difference = 0.27 D; P = 0.002). PALs caused a myopic shift in peripheral defocus. Superior myopic defocus was associated with less central myopia progression. These data support the continued investigation of optical designs that result in peripheral myopic defocus as a potential way to slow myopia progression. (ClinicalTrials.gov number, NCT00335049.).


Molecular characterization, expression, and in vivo analysis of LmexCht1: the chitinase of the human pathogen, Leishmania mexicana.
Chitinases have been implicated to be of importance in the life cycle development and transmission of a variety of parasitic organisms. Using a molecular approach, we identified and characterized the structure of a single copy LmexCht1-chitinase gene from the primitive trypanosomatid pathogen of humans, Leishmania mexicana. The LmexCht1 encodes an approximately 50 kDa protein, with well conserved substrate binding and catalytic domains characteristic of members of the chitinase-18 protein family. Further, we showed that LmexCht1 mRNA is constitutively expressed by both the insect vector (i.e. promastigote) and mammalian (i.e. amastigote) life cycle developmental forms of this protozoan parasite. Interestingly, however, amastigotes were found to secrete/release approximately >2-4-fold higher levels of chitinase activity during their growth in vitro than promastigotes. Moreover, a homologous episomal expression system was devised and used to express an epitope-tagged LmexCht1 chimeric construct in these parasites. Expression of the LmexCht1 chimera was verified in these transfectants by reverse transcription-PCR, Western blots, and indirect immunofluorescence analyses. Further, results of coupled immunoprecipitation/enzyme activity experiments demonstrated that the LmexCht1 chimeric protein was secreted/released by these transfected L. mexicana parasites and that it possessed functional chitinase enzyme activity. Such transfectants were also evaluated for their infectivity both in human macrophages in vitro and in two different strains of mice. Results of those experiments demonstrated that the LmexCht1 transfectants survived significantly better in human macrophages and also produced significantly larger lesions in mice than control parasites. Taken together, our results indicate that the LmexCht1-chimera afforded a definitive survival advantage to the parasite within these mammalian hosts. Thus, the LmexCht1 could potentially represent a new virulence determinant in the mammalian phase of this important human pathogen.


Acetylation-mediated transcriptional activation of the ETS protein ER81 by p300, P/CAF, and HER2/Neu.
The regulated expression of the ETS transcription factor ER81 is a prerequisite for normal development, and its dysregulation contributes to neoplasia. Here, we demonstrate that ER81 is acetylated by two coactivators/acetyltransferases, p300 and p300- and CBP-associated factor (P/CAF) in vitro and in vivo. Whereas p300 acetylates two lysine residues (K33 and K116) within the ER81 N-terminal transactivation domain, P/CAF targets only K116. Acetylation of ER81 not only enhances its ability to transactivate but also increases its DNA binding activity and in vivo half-life. Furthermore, oncogenic HER2/Neu, which induces phosphorylation and thereby activation of ER81, was less able to activate acetylation-deficient ER81 mutants, indicating that both acetyltransferase and protein kinase-specific regulatory mechanisms control ER81 activity. Importantly, HER2/Neu overexpression stimulates the ability of p300 to acetylate ER81, likely by inducing phosphorylation of p300 through the Ras-->Raf-->mitogen-activated protein kinase pathway. This represents a novel mechanism by which oncogenic HER2/Neu, Ras, or Raf may promote tumor formation by enhancing acetylation not only of ER81 but also of other downstream effector transcription factors as well as histones.


Holistic processing of faces: learning effects with Mooney faces.
The specialness of faces is seen in the face inversion effect, which disrupts the configural, but not the analytic, processing of faces. Mooney faces, which are processed holistically, allowed us to determine the contribution of holistic processing to the face inversion effect. As inverted Mooney faces are difficult to recognize as faces, we also included an intermediary training period for Mooney face recognition for half of the subjects. Early face-sensitive ERPs (N170 and P1) and P2 were measured. Behavioral data showed an increase in correct responses to inverted and upright Mooney faces after the learning phase for the experimental group. No effects were seen on P1. N170 latency did not vary with stimulus type before the intermediary phase, however, N170 amplitude was consistently larger for upright than inverted Mooney faces. After the intermediary exercise, N170 was delayed for inverted compared to upright Mooney faces. In contrast, for both groups of subjects P2 amplitude was larger for nonface stimuli, and P2 amplitude decreased after the intermediate task only for the subjects trained to recognize Mooney faces. As the usual inversion effect seen with photographic faces (delayed and larger N170) was not seen with Mooney faces, these data suggest that this effect on N170 is due to the recruitment of analytic processing. P2 reflected learning and a deeper processing of the stimuli that were not identifiable as faces.


Structural and Functional MRI Evidence for Distinct Medial Temporal and Prefrontal Roles in Context-dependent Relational Memory.
Declarative memory is supported by distributed brain networks in which the medial-temporal lobes (MTLs) and pFC serve as important hubs. Identifying the unique and shared contributions of these regions to successful memory performance is an active area of research, and a growing literature suggests that these structures often work together to support declarative memory. Here, we present data from a context-dependent relational memory task in which participants learned that individuals belonged in a single room in each of two buildings. Room assignment was consistent with an underlying contextual rule structure in which male and female participants were assigned to opposite sides of a building and the side assignment switched between buildings. In two experiments, neural correlates of performance on this task were evaluated using multiple neuroimaging tools: diffusion tensor imaging (Experiment 1), magnetic resonance elastography (Experiment 1), and functional MRI (Experiment 2). Structural and functional data from each individual modality provided complementary and consistent evidence that the hippocampus and the adjacent white matter tract (i.e., fornix) supported relational memory, whereas the ventromedial pFC/OFC (vmPFC/OFC) and the white matter tract connecting vmPFC/OFC to MTL (i.e., uncinate fasciculus) supported memory-guided rule use. Together, these data suggest that MTL and pFC structures differentially contribute to and support contextually guided relational memory.


Clinician barriers and facilitators to heart failure advance care plans: a systematic literature review and qualitative evidence synthesis.
Clinicians hesitate to engage with advance care planning (ACP) in heart failure. We aimed to identify the disease-specific barriers and facilitators for clinicians to engage with ACP. We searched Medline, Embase, CINAHL, PubMed, Scopus, the British Nursing Index, the Cochrane Library, the EPOC register, ERIC, PsycINFO, the Science Citation Index and the Grey Literature from inception to July 2018. We conducted the review according to Enhancing Transparency in Reporting the Synthesis of Qualitative Research (ENTREQ) guidelines. Two reviewers independently assessed original and empirical studies according to Critical Appraisal Skills Programme criteria. The SURE framework and thematic analysis were used to identify barriers and facilitators. Of 2308 articles screened, we reviewed the full text of 42 studies. Seventeen studies were included. The main barriers were lack of disease-specific knowledge about palliative care in heart failure, high emotional impact on clinicians when undertaking ACP and lack of multidisciplinary collaboration between healthcare professionals to reach consensus on when ACP is indicated. The main facilitators were being competent to provide holistic care when using ACP in heart failure, a patient taking the initiative of having an ACP conversation, and having the resources to deliver ACP at a time and place appropriate for the patient. Training healthcare professionals in the delivery of ACP in heart failure might be as important as enabling patients to start an ACP conversation. This twofold approach may mitigate against the high emotional impact of ACP. Complex interventions are needed to support clinicians as well as patients to engage with ACP.


Tissue residues of dietary cadmium in wood ducks.
One-week-old wood ducks were fed cadmium in diets containing 18 or 30% protein for a period of three months. Seven drakes from each group were sacrificed, and blood, brain, muscle, kidney, liver, and wing feather tissues were collected and analyzed for cadmium. Highest cadmium concentrations were found in the kidney, liver, and feather tissues; blood, muscle, and brain cadmium residue levels were undetectable. Except in the kidney tissue, protein level of the diet did not affect cadmium residue levels. For birds that were changed to a cadmium-free, high-protein ration at 13 weeks of age, regression analysis indicated a significant decrease in cadmium residue levels for the kidney tissue only. Growth, as measured by bodyweight at 13 weeks of age, was not affected by the various cadmium treatment levels. Histopathological examination revealed lesions in the kidney tissues of the 100 ppm cadmium treatment groups, which were more severe in those birds receiving the 18% protein diet. Vacuolation of the pancreatic acinar cells was observed in all groups, but tended to occur more frequently in the higher cadmium level groups.


Antibacterial activities of amphiphilic cyclic cell-penetrating peptides against multidrug-resistant pathogens.
Multidrug-resistant pathogens have become a major public health concern. There is a great need for the development of novel antibiotics with alternative mechanisms of action for the treatment of life-threatening bacterial infections. Antimicrobial peptides, a major class of antibacterial agents, share amphiphilicity and cationic structural properties with cell-penetrating peptides (CPPs). Herein, several amphiphilic cyclic CPPs and their analogues were synthesized and exhibited potent antibacterial activities against multidrug-resistant pathogens. Among all the peptides, cyclic peptide [R4W4] (1) showed the most potent antibacterial activity against methicillin-resistant Staphylococcus aureus [MRSA, exhibiting a minimal inhibitory concentration (MIC) of 2.67 μg/mL]. Cyclic [R4W4] and the linear counterpart R4W4 exhibited MIC values of 42.8 and 21.7 μg/mL, respectively, against Pseudomonas aeruginosa. In eukaryotic cells, peptide 1 exhibited the expected cell penetrating properties and showed >84% cell viability at a concentration of 15 μM (20.5 μg/mL) in three different human cell lines. Twenty-four hour time-kill studies evaluating [R4W4] with 2 times the MIC in combination with tetracycline demonstrated bactericidal activity at 4 and 8 times the MIC of tetracycline against MRSA (MIC = 0.5 μg/mL) and 2-8 times the MIC against Escherichia coli (MIC = 2 μg/mL). This study suggests that when amphiphilic cyclic CPPs are used in combination with an antibiotic such as tetracycline, they provide significant benefit against multidrug-resistant pathogens when compared with the antibiotic alone.


Effects of sphingolipid synthesis inhibition on cholesterol gallstone formation in C57BL/6J mice.
Sphingolipids play a very important role in cell membrane formation, signal transduction and plasma lipoprotein metabolism. The first rate-limiting step in the sphingolipid biosynthetic pathway is catalyzed by serine palmitoyltransferase (SPT), and myriocin is a potent and specific inhibitor of SPT. We investigated the impact of SPT inhibition on cholesterol gallstone formation in C57BL/6J mice. Three groups of eight-week-old C57BL/6J mice were utilized. Each group consisted of 20 mice; group A, B, and C were fed normal chow, lithogenic diet with phosphate buffered saline, and lithogenic diet with myriocin (0.3 mg/kg), respectively, for 6 weeks. The ceramide levels in both serum and bile were assessed by high performance liquid chromatography analysis. Protein expression of ERK, JNK and p38 in the extracted gallbladder were determined by Western-blot analysis. Myriocin treatment caused a significant decrease in the rate of cholesterol gallstone formation. The lithogenic diet mice (group B) showed the highest ceramide activities in both the serum and bile among all the tested groups and there was significant suppression of the ceramide levels in both the serum and bile of the myriocin-treated mice (group C, p < 0.05). Phosphorylation of p38 in the gallbladder was increased in the lithogenic-diet mice and the expression of phosphorylated p38 was significantly suppressed in the myriocin treated mice. SPT inhibition by myriocin suppressed gallstone formation and the levels of ceramide in both the serum and bile. p38 in the cellular signaling pathways might be associated with cholesterol gallstone formation.


The combined (Genesis) technique of radial keratotomy. A prospective, multicenter study. Refractive Keratoplasty Study Group.
A prospective, multicenter, clinical study to evaluate a standardized surgical (Genesis) protocol which includes combined-technique radial incisions in patients seeking reduction of their physiologic myopia. The combined incisions were designed to incorporate the safety of the centrifugal incision technique with the efficacy of the centripetal incision technique. A total of 375 eyes undergoing radial keratotomy procedures performed in six different clinical centers were analyzed. All procedures were performed in accordance with the Genesis nomograms. The Genesis protocol called for using preoperative screening pachymetry to guide central clear zone size selection, incising the thinnest corneal quadrant first, suturing corneal perforations, and discouraged more than one enhancement procedure, when indicated. Globe fixation technique served as a study variable. Mean follow-up was 6.2 months (range, 1.5-12 months). Mean residual cycloplegic refraction was -0.48 +/- 0.61 diopters (D) (range, -2.50 to +1.50 D); 92% of eyes were within 1 D of the planned goal of -0.50 D and 85% were within 1 D of emmetropia; 14% were myopic; and 1% was hyperopic by more than 1 D. Uncorrected visual acuity was 20/40 or better in 95% of eyes; the remaining 5% retaining myopic refractive errors. A single procedure was performed in 73% of eyes, and 99% received less than two enhancements. Of eyes with no enhancements, 97% had uncorrected visual acuity of 20/40 or better. One study eye (0.3%) had a two-line loss of spectacle visual acuity. There were no invasions of the central clear zone. Globe fixation was a significant predictor for enhancement incidence (P < 0.001) but not for perforation incidence (P = 0.06). Incision sequence was predictive for perforation incidence (P < 0.0002). The combined-technique of radial keratotomy, coupled with the Genesis surgical protocol, affords centrifugal incision safety with centripetal incision efficacy. The Genesis nomograms, with a built in refractive outcome goal of -0.50 D provide an acceptable degree of accuracy and predictability while guarding against hyperopic overcorrection.


Hip flexion contracture and diminished functional outcomes in cerebral palsy.
Hip flexion contracture (HFC) in the ambulatory child with cerebral palsy (CP) may impair function and lead to deteriorations in health-related quality of life. Furthermore, increasing HFC may lead to increasing disability. However, the association between passive range of motion and the measures of function and well-being is unclear. This study was designed to determine whether increasing HFC is associated with functional outcome. A total of 181 children, with an average age of 14.0 ± 10.2 years, were evaluated as part of a multicenter prospective data collection of patients with ambulatory CP. Measurements of HFC were recorded, and patients were evaluated using walking score from Gillette Functional Assessment Questionnaire (FAQ), Gross Motor Function Measure (GMFM), and the Pediatric Outcome Data Collection Instrument (PODCI). Patients were grouped on the basis of severity of HFC: group A, 0 to 14 degrees; group B, 15 to 29 degrees; and group C ≥ 30 degrees. Associations were examined using the Spearman correlation. There was an inverse association between degree of HFC and FAQ walking score (P<0.01, ρ=-0.25). Similarly, there was an inverse association between the degree of HFC and GMFM parts D (P<0.001, ρ=-0.31) and E (P<0.001, ρ=-0.32). Lastly, the PODCI domains of global function, mobility, and physical function also showed an inverse association with degree of HFC (P<0.001, ρ=-0.24). As surgeons treating children with CP, we often rely on joint measurements as an indirect measure of function. This study of children with ambulatory CP suggests that increased HFC from the physician's perspective is associated with deterioration in function from a patient and a therapist's perspective. Level II, prospective study.


Relationship between AgNOR proteins, Ki-67 antigen, p53 immunophenotype and differentiation markers in archival breast carcinomas.
The present study investigated (i) the relationship between standardised morphometric AgNOR parameters (argyrophilic nucleolar organiser region-associated proteins) and MIB1 growth fraction, and (ii) their correlation with immunohistochemical p53, sex steroid receptor status and histopathological differentiation grade in serial paraffin sections from 39 breast carcinomas. Ten sections were double-stained for AgNOR/MIB1. AgNOR parameters correlated significantly with MIB1 growth fraction and p53 protein expression. Significant inverse correlation was found between proliferation markers and oestrogen/progesterone receptor status and histopathological grade. AgNOR expression was significantly higher in cycling (MIB1 positive) tumour cells, than in resting (MIB1 negative) ones, however with exceptions. We conclude, that standardised AgNOR parameters correlate with markers of increased malignant potential in breast carcinomas. However, AgNORs seem to reflect proliferation independent cellular and nucleolar activity of tumour cells, as well. We recommend the use of standardised AgNOR analysis for obtaining sound results in routine paraffin sections.


Computational identification of microRNAs in apple expressed sequence tags and validation of their precise sequences by miR-RACE.
Thirty-one potential miRNAs that belong to 16 miRNA families were discovered from more than 324 000 EST sequences of apple (Malus domestica). In addition, precise sequences, especially terminal nucleotides of the 16 apple miRNAs (mdo-miRNAs) in 16 families were validated by miR-RACE, a newly developed method for the determination of the potential miRNAs predicted computationally. The expression of these 16 microRNAs could be detected in apple young leaf, old leaf, young stem, flower bud, flower and developing fruits by quantitative RT-PCR (qRT-PCR) and some of them showed tissue-specific expression. Fifty-six potential targets were identified for the 16 apple miRNAs, most of which were transcription factors that play important roles in apple development. Twelve target genes were experimentally verified by qRT-PCR, with some exhibiting different expression trends from their corresponding microRNAs, indicating the cleavage mode of miRNAs on their target genes.


Direct measurement of electric-field-induced birefringence in a polymer-stabilized blue-phase liquid crystal composite.
We demonstrate a method to directly measure the electric-field-induced birefringence of a polymer-stabilized blue-phase liquid crystal (PS-BPLC) composite. The induced birefringence follows the extended Kerr effect well and is approximately 3X the ordinary refractive index change. The measured data are validated by comparing the simulated and measured voltage-dependent transmittance with an in-plane switching cell. The impact of these results to the material optimization of emerging BPLC displays is discussed.


Getting more out of an incommensurately modulated structure: the example of K5Yb(MoO4)4.
A method based on the superspace approach is presented with the aim of generating a family of modular structures from a single incommensurately modulated structure. This approach based on the variation of the modulation vector q is applied to the generation of the K5Yb(MoO4)4, potassium ytterbium tetramolybdate, family of modular structures. The beta coefficient of the modulation vector q = betab* is a temperature-dependent variable which determines the modification. Our method gives a unified frame to describe and explain the three temperature-dependent phases of K5Yb(MoO4)4. Phase beta can be represented as a polytypic modification with 1/2b* < or = q < or = 2/3b*; phases gamma (q = 1/2b*) and alpha (q = 1b*) are the lowest and the highest temperature members of the K5Yb(MoO4)4 family, respectively.


White light induced covalent modification of graphene using a phenazine dye.
Herein, we report a novel strategy for a covalent modification of graphene nanoplatelets with photoactive dyes. The functionalization of the graphene surface was carried out using white light to photochemically generate phenazine radicals and the reaction progress was followed up spectrophotometrically. The characterization of the modified material was carried out using FTIR, XRD, UV-vis absorption, fluorescence, Raman spectroscopy and SEM imaging. The hybrid material has improved solubility, shows an optical band gap of 1.95 eV and is highly emissive in the visible wavelength region.


Screening for Atrial Fibrillation: A Report of the AF-SCREEN International Collaboration.
Approximately 10% of ischemic strokes are associated with atrial fibrillation (AF) first diagnosed at the time of stroke. Detecting asymptomatic AF would provide an opportunity to prevent these strokes by instituting appropriate anticoagulation. The AF-SCREEN international collaboration was formed in September 2015 to promote discussion and research about AF screening as a strategy to reduce stroke and death and to provide advocacy for implementation of country-specific AF screening programs. During 2016, 60 expert members of AF-SCREEN, including physicians, nurses, allied health professionals, health economists, and patient advocates, were invited to prepare sections of a draft document. In August 2016, 51 members met in Rome to discuss the draft document and consider the key points arising from it using a Delphi process. These key points emphasize that screen-detected AF found at a single timepoint or by intermittent ECG recordings over 2 weeks is not a benign condition and, with additional stroke factors, carries sufficient risk of stroke to justify consideration of anticoagulation. With regard to the methods of mass screening, handheld ECG devices have the advantage of providing a verifiable ECG trace that guidelines require for AF diagnosis and would therefore be preferred as screening tools. Certain patient groups, such as those with recent embolic stroke of uncertain source (ESUS), require more intensive monitoring for AF. Settings for screening include various venues in both the community and the clinic, but they must be linked to a pathway for appropriate diagnosis and management for screening to be effective. It is recognized that health resources vary widely between countries and health systems, so the setting for AF screening should be both country- and health system-specific. Based on current knowledge, this white paper provides a strong case for AF screening now while recognizing that large randomized outcomes studies would be helpful to strengthen the evidence base.


Lifetime exposure to traumatic and other stressful life events and hair cortisol in a multi-racial/ethnic sample of pregnant women.
We examined whether lifetime exposure to stressful and traumatic events alters hypothalamic-pituitary-adrenal (HPA) axis functioning, as indexed by hair cortisol, regardless of associated psychopathology, among pregnant women of different racial/ethnic backgrounds. 180 women provided hair samples for measurement of integrated cortisol levels throughout pregnancy and information regarding their lifetime exposure to stressful and traumatic life events. Results indicate that increased lifetime exposure to traumatic events was associated with significantly greater hair cortisol over the course of pregnancy. Similarly, greater lifetime exposure to stressful and traumatic events weighted by reported negative impact (over the previous 12 months) was associated with significantly greater hair cortisol during pregnancy. All analyses controlled for maternal age, education, body mass index (BMI), use of inhaled corticosteroids, race/ethnicity, and post-traumatic stress disorder (PTSD) and depressive symptoms. Following stratification by race/ethnicity, associations between stressful and traumatic life events and hair cortisol were found among Black women only. This is the first study to consider associations between lifetime stress exposures and hair cortisol in a sociodemographically diverse sample of pregnant women. Increased exposure to stressful and traumatic events, independent of PTSD and depressive symptoms, was associated with higher cortisol production, particularly in Black women. Future research should investigate the influence of such increased cortisol exposure on developmental outcomes among offspring.


HIV-1 Transmitted Drug Resistance Mutations in Newly Diagnosed Antiretroviral-Naive Patients in Turkey.
HIV-1 replication is rapid and highly error-prone. Transmission of a drug-resistant HIV-1 strain is possible and occurs within the HIV-1-infected population. In this study, we aimed to determine the prevalence of transmitted drug resistance mutations (TDRMs) in 1,306 newly diagnosed untreated HIV-1-infected patients from 21 cities across six regions of Turkey between 2010 and 2015. TDRMs were identified according to the criteria provided by the World Health Organization's 2009 list of surveillance drug resistance mutations. The HIV-1 TDRM prevalence was 10.1% (133/1,306) in Turkey. Primary drug resistance mutations (K65R, M184V) and thymidine analogue-associated mutations (TAMs) were evaluated together as nucleos(t)ide reverse transcriptase inhibitor (NRTI) mutations. NRTI TDRMs were found in 8.1% (107/1,306) of patients. However, TAMs were divided into three categories and M41L, L210W, and T215Y mutations were found for TAM1 in 97 (7.4%) patients, D67N, K70R, K219E/Q/N/R, T215F, and T215C/D/S mutations were detected for TAM2 in 52 (3.9%) patients, and M41L + K219N and M41L + T215C/D/S mutations were detected for the TAM1 + TAM2 profile in 22 (1.7%) patients, respectively. Nonnucleoside reverse transcriptase inhibitor-associated TDRMs were detected in 3.3% (44/1,306) of patients (L100I, K101E/P, K103N/S, V179F, Y188H/L/M, Y181I/C, and G190A/E/S) and TDRMs to protease inhibitors were detected in 2.3% (30/1,306) of patients (M46L, I50V, I54V, Q58E, L76V, V82A/C/L/T, N83D, I84V, and L90M). In conclusion, long-term and large-scale monitoring of regional levels of HIV-1 TDRMs informs treatment guidelines and provides feedback on the success of HIV-1 prevention and treatment efforts.


Hypoglycemia in Prader-Willi syndrome.
Although mouse models of Prader-Willi syndrome (PWS) suggest that hypoglycemia may be part of this syndrome, review of the literature shows little evidence that it is an issue in humans with PWS. Both adrenal and growth hormone deficiency can be seen in PWS, and both of these hormone deficiencies are associated with increased risk for hypoglycemia. We reviewed medical records for patients with PWS seen at the University of Florida Prader-Willi Program. Children were 2 months to 5 years of age. We recorded the presence, degree, and persistence of hypoglycemia, age of first occurrence, genetic diagnosis, and gestational age. Repeated hypoglycemia was determined if individuals had multiple blood glucose (BG) values <50 mg/dl before 1 month old, or BG levels <50 mg/dl once and additional BG values <70 mg/dl after 1 year of age. Of 95 patient charts reviewed, 12 (12.6%) had recorded hypoglycemia. Six of 12 patients with hypoglycemia had documented BG levels <40 mg/dl. Seven had their first episode of hypoglycemia within the first day of life. Of these seven individuals, five had BG <40 mg/dl. Repeated hypoglycemia was noted in 10 patients (83% of all patients with hypoglycemia). Only two with hypoglycemia had documented adrenal insufficiency. Our study suggests that infants with PWS may be predisposed to hypoglycemia from birth. Additional investigation is necessary to confirm our findings and define the cause of hypoglycemia. If the presence of hypoglycemia is confirmed, early detection and treatment may result in improved neurocognitive outcomes.


Activation of GCN2 kinase by ribosome stalling links translation elongation with translation initiation.
Ribosome stalling during translation has recently been shown to cause neurodegeneration, yet the signaling pathways triggered by stalled elongation complexes are unknown. To investigate these pathways we analyzed the brain of C57BL/6J-Gtpbp2(nmf205)(-/-) mice in which neuronal elongation complexes are stalled at AGA codons due to deficiencies in a tRNA(Arg)UCU tRNA and GTPBP2, a mammalian ribosome rescue factor. Increased levels of phosphorylation of eIF2α (Ser51) were detected prior to neurodegeneration in these mice and transcriptome analysis demonstrated activation of ATF4, a key transcription factor in the integrated stress response (ISR) pathway. Genetic experiments showed that this pathway was activated by the eIF2α kinase, GCN2, in an apparent deacylated tRNA-independent fashion. Further we found that the ISR attenuates neurodegeneration in C57BL/6J-Gtpbp2(nmf205)(-/-) mice, underscoring the importance of cellular and stress context on the outcome of activation of this pathway. These results demonstrate the critical interplay between translation elongation and initiation in regulating neuron survival during cellular stress.


Cost-effectiveness of Occupational Therapy in Older People: Systematic Review of Randomized Controlled Trials.
A systematic review of the cost-effectiveness of occupational therapy for older people was conducted. MEDLINE, CINAHL, Web of Science, PsycINFO, Cochrane Library, OT seeker and unpublished trials registers were searched. Reference lists of all potentially eligible studies were searched with no language restrictions. We included trial-based full economic evaluations that considered both costs and outcomes in occupational therapy for older people compared with standard care (i.e. other therapy) or no intervention. We reviewed each trial for methodological quality using the Cochrane risk of bias tool and assessed the quality of economic evaluations using a Drummond checklist. In the results of this review, we included five eligible studies (1-5) that were randomized controlled trials with high-quality economic evaluation. Two studies were full economic evaluations of interventions for fall prevention (1 and 2); two studies were full economic evaluations of preventive occupational therapy interventions (3 and 4; one was a comparison of an occupational therapy group with a social work group); one study was a full economic evaluation of occupational therapy for individuals with dementia (5). Two of the studies (one was preventive occupational therapy [3] and the other was occupational therapy for dementia [5]) found a significant effect and confirmed the cost-effectiveness of occupational therapy for older people compared with the control group. These studies found that occupational therapy for older people was clinically effective and cost-effective in comparison with standard care or other therapies. With reference to their clinical implication, these intervention studies (using a client-centred approach) suggested potentially cost-effective means to motivate clients to maintain their own health. However, this review has limitations because of the high heterogeneity of the reviewed studies on full economic evaluations of occupational therapy for older people. Future studies on the cost-effectiveness of occupational therapy in older people are strongly warranted. Copyright © 2015 John Wiley & Sons, Ltd.


Role for cyclin-dependent kinase 2 in mitosis exit.
Mitosis requires cyclin-dependent kinase (cdk) 1-cyclin B activity [1]. Exit from mitosis depends on the inactivation of the complex by the degradation of cyclin B [2]. Cdk2 is also active during mitosis [3, 4]. In Xenopus egg extracts, cdk2 is primarily in complex with cyclin E, which is stable [5]. At the end of mitosis, downregulation of cdk2-cyclin E activity is accompanied by inhibitory phosphorylation of cdk2 [6]. Here, we show that cdk2-cyclin E activity maintains cdk1-cyclin B during mitosis. At mitosis exit, cdk2 is inactivated prior to cdk1. The loss of cdk2 activity follows and depends upon an increase in protein kinase A (PKA) activity. Prematurely inactivating cdk2 advances the time of cyclin B degradation and cdk1 inactivation. Blocking PKA, instead, stabilizes cdk2 activity and inhibits cyclin B degradation and cdk1 inactivation. The stabilization of cdk1-cyclin B is also induced by a mutant cdk2-cyclin E complex that is resistant to inhibitory phosphorylation. P21-Cip1, which inhibits both wild-type and mutant cdk2-cyclin E, reverses mitotic arrest under either condition. Our findings indicate that the proteolysis-independent downregulation of cdk2 activity at the end of mitosis depends on PKA and is required to activate the proteolysis cascade that leads to mitosis exit.


A Novel Approach to Generate a Virtual Population of Human Coronary Arteries for In Silico Clinical Trials of Stent Design.
Goal: To develop a cardiovascular virtual population using statistical modeling and computational biomechanics. Methods: A clinical data augmentation algorithm is implemented to efficiently generate virtual clinical data using a real clinical dataset. An atherosclerotic plaque growth model is employed to 3D reconstructed coronary arterial segments to generate virtual coronary arterial geometries (geometrical data). Last, the combination of the virtual clinical and geometrical data is achieved using a methodology that allows for the generation of a realistic virtual population which can be used in in silico clinical trials. Results: The results show good agreement between real and virtual clinical data presenting a mean gof 0.1 ± 0.08. 400 virtual coronary arteries were generated, while the final virtual population includes 10,000 patients. Conclusions: The virtual arterial geometries are efficiently matched to the generated clinical data, both increasing and complementing the variability of the virtual population.


Conditional inactivation of Brca1, p53 and Rb in mouse ovaries results in the development of leiomyosarcomas.
Epithelial ovarian cancer (EOC) is thought to arise in part from the ovarian surface epithelium (OSE); however, the molecular events underlying this transformation are poorly understood. Germline mutations in the BRCA1 tumor suppressor gene result in a significantly increased risk of developing EOC and a large proportion of sporadic EOCs display some sort of BRCA1 dysfunction. To generate a model in which Brca1-mediated transformation can be studied, we previously inactivated Brca1 alone in murine OSE, which resulted in an increased accumulation of premalignant changes, but no tumor formation. In this study, we examined tumor formation in mice with conditionally expressed alleles of Brca1, p53 and Rb, alone or in combination. Intrabursal injection of adenovirus expressing Cre recombinase to inactivate p53 resulted in tumors in 100% of mice. Tumor progression was accelerated in mice with concomitant inactivation of Brca1 and p53, but not Rb and p53. Immunohistologic analyses classified the tumors as leiomyosarcomas that may be arising from the ovarian bursa. Brca1 inactivation in primary cultures of murine OSE cells led to a suppression of proliferation that could be rescued by concomitant inactivation of p53 and/or Rb. Brca1-deficient OSE cells displayed an increased sensitivity to the DNA damaging agent cisplatin, and this effect could be modulated by inactivation of p53 and/or Rb. These results indicate that Brca1 deficiency can accelerate tumor development and alter the sensitivity of OSE cells to chemotherapeutic agents. Intrabursal delivery of adenovirus intended to alter gene expression in the ovarian surface epithelium may, in some strains of mice, result in more rapid transformation of adjacent cells, resulting in leiomyosarcomas.


Spatial segregation in eastern North Pacific skate assemblages.
Skates (Rajiformes: Rajoidei) are common mesopredators in marine benthic communities. The spatial associations of individual species and the structure of assemblages are of considerable importance for effective monitoring and management of exploited skate populations. This study investigated the spatial associations of eastern North Pacific (ENP) skates in continental shelf and upper continental slope waters of two regions: central California and the western Gulf of Alaska. Long-term survey data were analyzed using GIS/spatial analysis techniques and regression models to determine distribution (by depth, temperature, and latitude/longitude) and relative abundance of the dominant species in each region. Submersible video data were incorporated for California to facilitate habitat association analysis. We addressed three main questions: 1) Are there regions of differential importance to skates?, 2) Are ENP skate assemblages spatially segregated?, and 3) When skates co-occur, do they differ in size? Skate populations were highly clustered in both regions, on scales of 10s of kilometers; however, high-density regions (i.e., hot spots) were segregated among species. Skate densities and frequencies of occurrence were substantially lower in Alaska as compared to California. Although skates are generally found on soft sediment habitats, Raja rhina exhibited the strongest association with mixed substrates, and R. stellulata catches were greatest on rocky reefs. Size segregation was evident in regions where species overlapped substantially in geographic and depth distribution (e.g., R. rhina and Bathyraja kincaidii off California; B. aleutica and B. interrupta in the Gulf of Alaska). Spatial niche differentiation in skates appears to be more pronounced than previously reported.


Determinants of Health Care Services Utilization among First Generation Afghan Migrants in Istanbul.
There is insufficient empirical evidence on the correlates of health care utilization of irregular migrants currently living in Turkey. The aim of this study was to identify individual level determinants associated with health service and medication use. One hundred and fifty-five Afghans completed surveys assessing service utilization including encounters with primary care physicians and outpatient specialists in addition to the use of prescription and nonprescription medicines. Multivariate logistic regression analyses were employed to examine associations between service use and a range of predisposing, enabling, and perceived need factors. Health services utilization was lowest for outpatient specialists (20%) and highest for nonprescription medications (37%). Female gender and higher income predicted encounters with primary care physicians. Income, and other enabling factors such as family presence in Turkey predicted encounters with outpatient specialists. Perceived illness-related need factors had little to no influence on use of services; however, asylum difficulties increased the likelihood for encounters with primary care physicians, outpatient services, and the use of prescription medications. This study suggests that health services use among Afghan migrants in Turkey is low considering the extent of their perceived illness-related needs, which may be further exacerbated by the precarious conditions in which they live.


Fine mapping of probes in the adenomatous polyposis coli region of chromosome 5 by in situ hybridization.
The gene for adenomatous polyposis coli has been localized to 5q21-22. We have mapped six probes from this region using isotopic or nonisotopic in situ hybridization. Using tritium-labeled probes we localized II227 (D5S37) to 5q14-15 and ECB27 (D5S98) to 5q21. Following hybridization with biotin-labeled probes, the positions of signals along the chromosomes were measured as fractional length relative to the length of the chromosome arm from centromere to qter (FLcen-qter). Ninety-five percent confidence limits, compared with standard karyotypes, provided the corresponding band localization. By this method we localized Cllpll (D5S71) to FLcen-qter 0.407-0.452 (5q21.1-21.3), ECB27 to FLcen-qter 0.426-0.473 (5q21.3), YN5.48 (D5S81) to FLcen-qter 0.459-0.496 (5q21.3-22.2), and ECB134 (D5S97) to FLcen-qter 0.509-0.533 (5q22.3-23.1). ECB220 had three sites of hybridization, a major site at FLcen-qter 0.460-0.492 (5q21.3-22.1) and minor sites at FLcen-qter 0.299-0.339 (5q14.3-15) and FLcen-qter 0.629-0.691 (5q23.3-31.2). We have shown that the chromosome 5 breakpoint in a t(5;15) translocation from a patient with Gardner's syndrome (GM03314) is between Cllpll and ECB27. Linkage data are presented suggesting that ECB27 is located on the same side of the APC locus as II227. These and published results including data on several constitutional deletions (M, SD, and brothers PW and ND) give a probable order of [cen] - [II227, proximal SD breakpoint] - [Cllpll] - [proximal PW/ND, M breakpoint(s), GM03314 breakpoint] - [ECB27] - [APC] - [YN5.48] - [distal PW/ND breakpoint] - [ECB134] - [distal M breakpoint] - [qter]. The major site of ECB220 appears to be between ECB27 and the distal PW/ND breakpoint; the distal SD breakpoint is distal to YN5.48.


Food-borne trematode infections of humans in the United States of America.
This review examines the literature on imported (allochthonous) and local (autochthonous) cases of food-borne trematode (FBT) infections in the United States of America (USA) from 1890 to 2009. Most of the literature is concerned with imported cases of the opisthorchiids Clonorchis sinensis and Opisthorchis viverrini. These flukes cause serious pathology in the liver and biliary system of humans. Chronic cases may induce liver (hepatocarcinoma) and bile duct (cholangiocarcinoma) cancers in humans. Clonorchiasis and opithorchiasis are preventable diseases that can be avoided by eating properly cooked freshwater fish products. Several species of lung flukes in the genus Paragonimus are local or imported FBT in the USA. The endemic cycle occurs in the USA with various local snails and crustaceans serving as intermediate hosts. Paragonimids are acquired when humans eat raw or improperly cooked freshwater crustaceans containing metacercarial cysts. Infection can cause severe lung disease and the symptoms of paragonimiasis often mimic those of tuberculosis and other non-helminthic diseases. Paragonimiasis can be avoided by not eating raw or improperly cooked shellfish. The liver fluke Fasciola hepatica can be acquired by eating raw or uncooked vegetation. The cycle exists in the USA involving local snails and aquatic vegetation. Although some cases are local, most are imported by travelers or immigrants. Fascioliasis can cause serious liver and biliary diseases in humans and consumption of tainted vegetation should be avoided. Lesser known FBT have been reported in the USA including species of Alaria, echinostomids, heterophyids, troglotrematids, and a self-induced infection of Plagiorchis. Treatment of the FBT mentioned in this review consists of various regimens of praziquantel, except for F. hepatica where the drug of choice is triclabendazole.


Cytotoxic effects of cuphiin D1 on the growth of human cervical carcinoma and normal cells.
Cuphiin D1 (CD1), macrocyclic hydrolyzable tannin isolated from Cuphea hyssopifolia, has been shown to exert an antitumor effect both in vitro and in vivo. Furthermore, CD1 significantly inhibited the growth of the human cervical carcinoma, i.e. HeLa, cells and showed less cytotoxicity to normal primary-cultured cervical fibroblasts. In this study, we explored the cytotoxic mechanism of CD1 on HeLa cells. The cytotoxic effects of CD1 showed dose-dependency at 3.15-100 micrograms/ml on HeLa for 12, 24, 48 and 72 hours and with an IC50 value at 14.2 micrograms/ml for 48 hours. However, the IC50 value of CD1 in primary-cultured normal cervical fibroblasts was 74.5 micrograms/ml. Therefore, the selectivity shown by CD1 is ascribed to differences in growth speeds between normal and tumor cells. HeLa cells treated with 50 micrograms/ml CD1 for 24 hours exhibited chromatin condensation, indicating the occurrence of apoptosis. Flow cytometric analysis demonstrated the presence of apoptotic cells with low DNA content among HeLa cells. CD1 also caused DNA fragmentation and inhibited Bcl-2, pro-caspase 3, and inactived PARP expression in HeLa cells. These results suggest that the inhibition of Bcl-2 expression in HeLa cells might account for the mechanism of CD1-induced apoptosis.


Esophageal clearance patterns in normal older adults as documented with videofluoroscopic esophagram.
Normal esophageal bolus transport in asymptomatic healthy older adults has not been well defined, potentially leading to ambiguity in differentiating esophageal swallowing patterns of dysphagic and healthy individuals. This pilot study of 24 young (45-64 years) and old (65+years) men and women was designed to assess radiographic esophageal bolus movement patterns in healthy adults using videofluoroscopic recording. Healthy, asymptomatic adults underwent videofluoroscopic esophagram to evaluate for the presence of ineffective esophageal clearance, namely, intraesophageal stasis and intraesophageal reflux. Intraesophageal stasis and intraesophageal reflux were visualized radiographically in these normal subjects. Intraesophageal stasis occurred significantly more frequently with semisolid (96%) compared with liquid (16%) barium, suggesting that a variety of barium consistencies, as opposed to only the traditional fluids, would better define the spectrum of esophageal transport. Intraesophageal reflux was observed more frequently in older males than in their younger counterparts. The rates of intraesophageal stasis and intraesophageal reflux were potentially high given that successive bolus presentations were spaced 10 seconds apart. These findings suggest a need for a more comprehensive definition regarding the range of normal esophageal bolus transport to (a) prevent misdiagnosis of dysphagia and (b) to enhance generalization to functional eating, which involves solid foods in addition to liquids.


Stimulus coding in human associative learning: flexible representations of parts and wholes.
An enduring theme for theories of associative learning is the problem of explaining how configural discriminations--ones in which the significance of combinations of cues is inconsistent with the significance of the individual cues themselves-are learned. One approach has been to assume that configurations are the basic representational form on which associative processes operate, another has tried in contrast to retain elementalism. We review evidence that human learning is representationally flexible in a way that challenges both configural and elemental theories. We describe research showing that task demands, prior experience, instructions, and stimulus properties all influence whether a particular problem is solved configurally or elementally. Lines of possible future theory development are discussed.


Reduction in sympathetic activity after long-term CPAP treatment in sleep apnoea: cardiovascular implications.
Twelve patients with severe obstructive sleep apnoea were included in an open, long-term, prospective, follow-up study addressing the effects of nasal continuous positive airway pressure (CPAP) on sympathetic activity, cardiac structure and blood pressure. Plasma norepinephrine (P-NE) (daytime at rest), daytime and night-time urinary excretion of NE (U-NE), vanylmandelic acid and metanephrines, together with 24 h noninvasive blood pressure (BP) recording and Doppler-echocardiography, were assessed before and after a mean of 20.5 (range 14-26) months of CPAP. Average self-reported use of CPAP was 89% (range 65-100%) of time spent in bed. Resting daytime P-NE ranged 0.35-0.83 ng.ml-1, which is elevated compared to healthy controls. Only night-time U-NE, mean daytime BP and average 24 h BP were related to severity of OSA. Night-time metanephrine was related to daytime and night-time diastolic, as well as night-time systolic, BP. Left ventricular mass index (LVMI) at baseline was correlated to daytime systolic BP and P-NE. Long-term CPAP treatment reduced biochemical markers of sympathetic activity. P-NE decreased by approximately 50%, and daytime and night-time vanylmandelic acid and metanephrine by 32-54%. In contrast, there were no overall reductions in BP or LVMI. It is concluded that obstructive sleep apnoea is associated with high sympathetic activity both during sleep and waking periods. Urinary metanephrine excretion seemed to reflect blood pressure, but neither daytime nor night-time catecholamine excretion was directly related to disease severity in patients with severe obstructive sleep apnoea.(ABSTRACT TRUNCATED AT 250 WORDS)


HIV Rev-isited.
The human immunodeficiency virus type 1 (HIV-1) proteome is expressed from alternatively spliced and unspliced genomic RNAs. However, HIV-1 RNAs that are not fully spliced are perceived by the host machinery as defective and are retained in the nucleus. During late infection, HIV-1 bypasses this regulatory mechanism by expression of the Rev protein from a fully spliced mRNA. Once imported into the nucleus, Rev mediates the export of unprocessed HIV-1 RNAs to the cytoplasm, leading to the production of the viral progeny. While regarded as a canonical RNA export factor, Rev has also been linked to HIV-1 RNA translation, stabilization, splicing and packaging. However, Rev's functions beyond RNA export have remained poorly understood. Here, we revisit this paradigmatic protein, reviewing recent data investigating its structure and function. We conclude by asking: what remains unknown about this enigmatic viral protein?


Dissociation between the impact of evidence on eye movement target choice and confidence judgements.
It has been suggested that the evidence used to support a decision to move our eyes and the confidence we have in that decision are derived from a common source. Alternatively, confidence may be based on further post-decisional processes. In three experiments, we examined this. In Experiment 1, participants chose between two targets on the basis of varying levels of evidence (i.e., the direction of motion coherence in a random dot kinematogram). They indicated this choice by making a saccade to one of two targets and then indicated their confidence. Saccade trajectory deviation was taken as a measure of the inhibition of the non-selected target. We found that as evidence increased so did confidence and deviations of saccade trajectory away from the non-selected target. However, a correlational analysis suggested they were not related. In Experiment 2, an option to opt-out of the choice was offered on some trials if choice proved too difficult. In this way, we isolated trials on which confidence in target selection was high (i.e., when the option to opt-out was available but not taken). Again saccade trajectory deviations were found not to differ in relation to confidence. In Experiment 3, we directly manipulated confidence, such that participants had high or low task confidence. They showed no differences in saccade trajectory deviations. These results support post-decisional accounts of confidence: evidence supporting the decision to move the eyes is reflected in saccade control, but the confidence that we have in that choice is subject to further post-decisional processes.


SnoRNA U50 levels are regulated by cell proliferation and rRNA transcription.
rRNA post transcriptional modifications play a role in cancer development by affecting ribosomal function. In particular, the snoRNA U50, mediating the methylation of C2848 in 28S rRNA, has been suggested as a potential tumor suppressor-like gene playing a role in breast and prostate cancers and B-cell lymphoma. Indeed, we observed the downregulation of U50 in colon cancer cell lines as well as tumors. We then investigated the relationship between U50 and proliferation in lymphocytes stimulated by phytohemagglutinin (PHA) and observed a strong decrease in U50 levels associated with a reduced C2848 methylation. This reduction was due to an alteration of U50 stability and to an increase of its consumption. Indeed, the blockade of ribosome biogenesis induced only an early decrease in U50 followed by a stabilization of U50 levels when ribosome biogenesis was almost completely blocked. Similar results were found with other snoRNAs. Lastly, we observed that U50 modulation affects ribosome efficiency in IRES-mediated translation, demonstrating that changes in the methylation levels of a single specific site on 28S rRNA may alter ribosome function. In conclusion, our results link U50 to the cellular proliferation rate and ribosome biogenesis and these findings may explain why its levels are often greatly reduced in cancers.


Internet Use Patterns, Internet Addiction, and Psychological Distress Among Engineering University Students: A Study from India.
Internet addiction (IA) among university engineering students and its association with psychological distress can impact their educational progress, academic competence, and long-term career goals. Thus, there is a need to investigate the IA among engineering students. This study was a first such attempt to explore internet use behaviors, IA, among a large group of engineering students from India, and its association with psychological distress primarily depressive symptoms. One thousand eighty six engineering students aged 18-21 years pursuing bachelors in engineering from the south Indian city of Mangalore participated in the study. The socio-educational and internet use behaviors data sheet was used to gather demographic information and patterns of internet use, Internet Addiction Test (IAT) was utilized to assess IA, and Self-Report Questionnaire (SRQ-20) assessed psychological distress primarily depressive symptoms. Among the total N = 1086, 27.1% of engineering students met criterion for mild addictive internet use, 9.7% for moderate addictive internet use, and 0.4% for severe addiction to internet. IA was higher among engineering students who were male, staying in rented accommodations, accessed internet several times a day, spent more than 3 h per day on internet, and had psychological distress. Gender, duration of use, time spent per day, frequency of internet use, and psychological distress (depressive symptoms) predicted IA. A substantial proportion of engineering students have IA which can be detrimental for their educational progress in university studies and long-term career goals. Early identification and management of IA and psychological distress among engineering students is crucial.


Pathogenic role of the Wnt signaling pathway activation in laser-induced choroidal neovascularization.
Choroidal neovascularization (CNV) is a severe complication of AMD. The Wnt signaling pathway has been shown to mediate angiogenesis. The purpose of this study was to investigate the pathogenic role of the Wnt pathway in CNV and explore the therapeutic potential of a novel Wnt signaling inhibitor in CNV. Adult rats and mice were photocoagulated using diode laser to induce CNV. On the same day, the animals were intravitreally injected with a monoclonal antibody (Mab2F1) blocking LRP6 or nonspecific mouse IgG. The Wnt signaling activation and target gene expression in the eyecup were determined by Western blot analysis. Fundus angiography was used to examine leakage from the laser lesion. CNV areas were measured on choroidal flatmount using FITC-dextran. Levels of Wnt pathway components and Wnt target gene expression were elevated in both laser-induced CNV rat and mouse eyecups, suggesting activation of the Wnt pathway. Significant suppression of Wnt signaling was observed in the Mab2F1 treatment group. Mab2F1 decreased vascular leakage from CNV lesions and reduced the neovascular area in laser-induced CNV rats. Mab2F1 inhibited the hypoxia-induced activation of Wnt signaling in cultured RPE cells. Mab2F1 also ameliorated retinal inflammation and vascular leakage in the eyecups of very low-density lipoprotein receptor knockout mice, a model of subretinal neovascularization. The Wnt pathway is activated in the laser-induced CNV models and plays a pathogenic role in CNV. Blockade of Wnt signaling using an anti-LRP6 antibody has therapeutic potential in CNV.


Differential responses of epithelial cells from urinary and biliary tract to eggs of Schistosoma haematobium and S. mansoni.
Chronic urogenital schistosomiasis can lead to squamous cell carcinoma of the bladder. The International Agency for Research on Cancer classifies the infection with S. haematobium as a group 1 carcinogen, a definitive cause of cancer. By contrast, hepatointestinal schistosomiasis due to the chronic infection with S. mansoni or S. japonicum associated with liver periportal fibrosis, does not apparently lead to malignancy. The effects of culturing human epithelial cells, HCV29, established from normal urothelium, and H69, established from cholangiocytes, in the presence of S. haematobium or S. mansoni eggs were investigated. Cell growth of cells co-cultured with schistosome eggs was monitored in real time, and gene expression analysis of oncogenesis, epithelial to mesenchymal transition and apoptosis pathways was undertaken. Schistosome eggs promoted proliferation of the urothelial cells but inhibited growth of cholangiocytes. In addition, the tumor suppressor P53 pathway was significantly downregulated when exposed to schistosome eggs, and downregulation of estrogen receptor was predicted in urothelial cells exposed only to S. haematobium eggs. Overall, cell proliferative responses were influenced by both the tissue origin of the epithelial cells and the schistosome species.


Critical limb ischemia: an update for interventional radiologists.
Critical limb ischemia (CLI) is a growing epidemic with bleak patient outcomes. A variety of treatment modalities have been adopted to address CLI based on comorbidities, life expectancy, and the nature of the arterial disease. With advances in technology and treatment strategies, the clinical outcomes of CLI patients have significantly improved over recent years. However, despite progress, patency rates of both surgical and endovascular interventions, limb-salvage and amputation rates are still dismal. We review the epidemiology, treatment strategies, imaging modalities, and the microcirculation aspect of CLI.


MicroRNAs as sentinels and protagonists of carotid artery thromboembolism.
Stroke is the leading cause of serious disability in the world and a large number of ischemic strokes are due to thromboembolism from unstable carotid artery atherosclerotic plaque. As it is difficult to predict plaque rupture and surgical treatment of asymptomatic disease carries a risk of stroke, carotid disease continues to present major challenges with regard to clinical decision-making and revascularization. There is therefore an imminent need to better understand the molecular mechanisms governing plaque instability and rupture, as this would allow for the development of biomarkers to identify at-risk asymptomatic carotid plaque prior to disease progression and stroke. Further, it would aid in creation of therapeutics to stabilize carotid plaque. MicroRNAs (miRNAs) have been implicated as key protagonists in various stages of atherosclerotic plaque initiation, development and rupture. Notably, they appear to play a crucial role in carotid artery thromboembolism. As the molecular pathways governing the role of miRNAs are being uncovered, we are learning that their involvement is complex, tissue- and stage-specific, and highly selective. Notably, miRNAs can be packaged and secreted in extracellular vesicles (EVs), where they participate in cell-cell communication. The measurement of EV-encapsulated miRNAs in the circulation may inform disease mechanisms occurring in the plaque itself, and therefore may serve as sentinels of unstable plaque as well as therapeutic targets.


Mutational analysis of transmembrane regions 3 and 4 of SecY, a central component of protein translocase.
The SecYEG heterotrimeric membrane protein complex functions as a channel for protein translocation across the Escherichia coli cytoplasmic membrane. SecY is the central subunit of the SecYEG complex and contains 10 transmembrane segments (TM1 to TM10). Previous mutation studies suggested that TM3 and TM4 are particularly important for SecY function. To further characterize TM3 and TM4, we introduced a series of cysteine-scanning mutations into these segments. With one exception (an unstable product), all the mutant proteins complemented the cold-sensitive growth defect of the secY39 mutant. A combination of this secY mutation and the secG deletion resulted in synthetic lethality, and the TM3 and TM4 SecY cysteine substitution mutations were examined for their ability to complement this lethality. Although they were all positive for complementation, some of the complemented cells exhibited significant retardation of protein export. The substitution-sensitive residues in TM3 can be aligned to one side of the alpha-helix, and those in TM4 revealed a tendency for residues closer to the cytosolic side of the membrane to be more severely affected. Disulfide cross-linking experiments identified a specific contact point for TM3 and SecG TM2 as well as for TM4 and SecG TM1. Thus, although TM3 and TM4 do not contain any single residue that is absolutely required, they include functionally important helix surfaces and specific contact points with SecG. These results are discussed in light of the structural information available for the SecY complex.


Failure at the top: How power undermines collaborative performance.
All too commonly, we see groups of leaders fail to accomplish their stated goals when working together--legislators who cannot agree on a bill, heads of state who cannot draft meaningful environmental policy, or boards of trustees who make disastrous decisions for their school. The current research examines whether groups of leaders fail as often as they do in part because of the power each leader is accustomed to possessing among his or her constituents. In multiple studies we found that high power individuals, when working together in groups, performed worse than did other groups: individuals randomly assigned power in an initial task were less creative when they then worked together in groups on a subsequent task (Studies 1A and 4). Individuals with higher power who worked together in groups were also less likely to reach agreement on a difficult negotiation task, whether these groups comprised actual executives from an extant organization (Study 2) or participants randomly assigned power in the laboratory (Study 3). Mediation analyses suggest that groups of high power individuals performed worse because they fought over their relative status in the group, were less focused on the task, and shared information with each other less effectively. However, high power individuals were more effective when working on tasks that required less coordination: they were more creative (Studies 1B and 4) and persisted longer on a difficult task than other groups. Therefore, group processes are the key problem for groups of high power individuals when they work together.


Timing Matters? Learning of Complex Spatiotemporal Sequences in Left-hemisphere Stroke Patients.
During rehabilitation after stroke motor sequence learning is of particular importance because considerable effort is devoted to (re)acquiring lost motor skills. Previous studies suggest that implicit motor sequence learning is preserved in stroke patients but were restricted to the spatial dimension, although the timing of single action components is as important as their spatial order. As the left parietal cortex is known to play a critical role in implicit timing and spatiotemporal integration, in this study we applied an adapted version of the SRT task designed to assess both spatial (different stimulus locations) and temporal (different response-stimulus intervals) aspects of motor learning to 24 right-handed patients with a single left-hemisphere (LH) stroke and 24 age-matched healthy controls. Implicit retrieval of sequence knowledge was tested both at Day 1 and after 24 hr (Day 2). Additionally, voxel-based lesion symptom mapping was used to investigate the neurobiological substrates of the behavioral effects. Although LH stroke patients showed a combined spatiotemporal learning effect that was comparable to that observed in controls, LH stroke patients did not show learning effects for the learning probes in which only one type of sequence information was maintained whereas the other one was randomized. Particularly on Day 2, patients showed significantly smaller learning scores for these two learning probes than controls. Voxel-based lesion symptom mapping analyses revealed for all learning probes that diminished learning scores on Day 2 were associated with lesions of the striatum. This might be attributed to its role in motor chunking and offline consolidation as group differences occurred on Day 2 only. The current results suggest that LH stroke patients rely on multimodal information (here: temporal and spatial information) when retrieving motor sequence knowledge and are very sensitive to any disruption of the learnt sequence information as they seem to build very rigid chunks preventing them from forming independent spatial and temporal sequence representations.


Risk factors associated with delayed discharge following robotic assisted surgery for gynecologic malignancy.
The risk factors for extended length of stay (LOS) have not been examined in a cohort of patients with complex social and medical barriers who undergo robotic assisted (RA) surgery for gynecologic malignancies. We sought to identify those patients with a LOS > 24 h after robotic surgery and the risk factors associated with delayed discharge. Then we aimed to develop a predictive model for clinical care and identify modifiable pre-operative risk factors. After IRB approval, data was abstracted from medical records of all patients with a gynecologic malignancy who underwent a RA laparoscopic surgery from 2010 to 2015. Univariable and multivariable logistic regression was performed to identify independent risk factors associated with delayed discharge defined as LOS > 24 h. A multi-variable logistic regression model was performed using a stepwise backward selection for the final prediction model. All testing was two-sided and a p-value < 0.05 was considered statistically significant. Of the 406 eligible and evaluable patients, 194 (48%) had a LOS > 24 h. Age ≥ 60 years, a higher usage of narcotic medication, a longer surgical time, and a larger estimated blood loss were all associated with LOS > 24 h (p < 0.05). Many of these women had a social work consultation and went home with home care services despite no surgical or post-operative complications. Our prediction model has the potential to correctly classified 75% of the patients discharged within 24 h. The development of a pre-hospitalization risk stratification and anticipating the possible need for home care services pre-operatively shows promise as a strategy to decrease LOS in patients classified as high-risk. These findings warrant prospective validation through the use of this prediction model in our institution.


Neuronal low-density lipoprotein receptor-related protein 1 binds and endocytoses prion fibrils via receptor cluster 4.
For infectious prion protein (designated PrP(Sc)) to act as a template to convert normal cellular protein (PrP(C)) to its distinctive pathogenic conformation, the two forms of prion protein (PrP) must interact closely. The neuronal receptor that rapidly endocytoses PrP(C) is the low-density lipoprotein receptor-related protein 1 (LRP1). We show here that on sensory neurons LRP1 is also the receptor that binds and rapidly endocytoses smaller oligomeric forms of infectious prion fibrils, and recombinant PrP fibrils. Although LRP1 binds two molecules of most ligands independently to its receptor clusters 2 and 4, PrP(C) and PrP(Sc) fibrils bind only to receptor cluster 4. PrP(Sc) fibrils out-compete PrP(C) for internalization. When endocytosed, PrP(Sc) fibrils are routed to lysosomes, rather than recycled to the cell surface with PrP(C). Thus, although LRP1 binds both forms of PrP, it traffics them to separate fates within sensory neurons. The binding of both to ligand cluster 4 should enable genetic modification of PrP binding without disrupting other roles of LRP1 essential to neuronal viability and function, thereby enabling in vivo analysis of the role of this interaction in controlling both prion and LRP1 biology.