Stereoselective cis-Vinylcyclopropanation via a Gold(I)-Catalyzed Retro-Buchner Reaction under Mild Conditions.
A highly stereoselective gold(I)-catalyzed cis-vinylcyclopropanation of alkenes has been developed. Allylic gold carbenes, generated via a retro-Buchner reaction of 7-alkenyl-1,3,5-cycloheptatrienes, react with alkenes to form vinylcyclopropanes. The gold(I)-catalyzed retro-Buchner reaction of these substrates proceeds by simple heating at a temperature much lower than that required for the reaction of 7-aryl-1,3,5-cycloheptatrienes (75 °C vs 120 °C). A newly developed Julia-Kocienski reagent enables the synthesis of the required cycloheptatriene derivatives in one step from readily available aldehydes or ketones. On the basis of mechanistic investigations, a stereochemical model for the cis selectivity was proposed. An unprecedented gold-catalyzed isomerization of cis- to trans-cyclopropanes has also been discovered and studied by DFT calculations.


The Pseudomonas syringae pv. tomato type III effector HopM1 suppresses Arabidopsis defenses independent of suppressing salicylic acid signaling and of targeting AtMIN7.
Pseudomonas syringae pv tomato strain DC3000 (Pto) delivers several effector proteins promoting virulence, including HopM1, into plant cells via type III secretion. HopM1 contributes to full virulence of Pto by inducing degradation of Arabidopsis proteins, including AtMIN7, an ADP ribosylation factor-guanine nucleotide exchange factor. Pseudomonas syringae pv phaseolicola strain NPS3121 (Pph) lacks a functional HopM1 and elicits robust defenses in Arabidopsis thaliana, including accumulation of pathogenesis related 1 (PR-1) protein and deposition of callose-containing cell wall fortifications. We have examined the effects of heterologously expressed HopM1Pto on Pph-induced defenses. HopM1 suppresses Pph-induced PR-1 expression, a widely used marker for salicylic acid (SA) signaling and systemic acquired resistance. Surprisingly, HopM1 reduces PR-1 expression without affecting SA accumulation and also suppresses the low levels of PR-1 expression apparent in SA-signaling deficient plants. Further, HopM1 enhances the growth of Pto in SA-signaling deficient plants. AtMIN7 contributes to Pph-induced PR-1 expression. However, HopM1 fails to degrade AtMIN7 during Pph infection and suppresses Pph-induced PR-1 expression and callose deposition in wild-type and atmin7 plants. We also show that the HopM1-mediated suppression of PR-1 expression is not observed in plants lacking the TGA transcription factor, TGA3. Our data indicate that HopM1 promotes bacterial virulence independent of suppressing SA-signaling and links TGA3, AtMIN7, and other HopM1 targets to pathways distinct from the canonical SA-signaling pathway contributing to PR-1 expression and callose deposition. Thus, efforts to understand this key effector must consider multiple targets and unexpected outputs of its action.


Interrelationship of emotional stability, hassles, uplifts, coping and stress-related symptoms in Swedish female and male military veterans.
Drawing on previous research, two hypotheses were tested: (1) the higher the frequency of daily uplifts and use of functional coping strategies, and the lower the frequency of daily hassles and use of dysfunctional coping strategies, the lower the prevalence of stress-related symptoms will be, and vice versa; and (2) the direct relationship between the personality dimension emotional stability and stress-related symptoms, will be moderated by daily hassles, daily uplifts and coping processes. A quantitative test of a qualitatively developed model was performed. A questionnaire was sent to all Swedish military veterans who had served in the period 2011-2015 and 1859 individuals (1,614 men and 199 women, 46 individuals did not mark gender) responded (40.5% total response rate). All analyses were made separately for men and women. Comparisons between theoretically favorable and unfavorable profiles across the model variables, daily uplifts, daily hassles, functional coping and dysfunctional coping (based on a cluster analysis), showed considerable differences regarding the prevalence of stress-related symptoms as predicted by the model and supporting the first hypothesis. Regression and moderation analyses yielded limited support for the second hypothesis. As predicted, female veterans reported a higher frequency of physical, emotional and cognitive stress-related symptoms than male veterans. The main conclusion is that the theoretical model stood up well when empirically tested and offers a promising approach to future studies on everyday stress and health. The results contribute with new knowledge of military veterans compared to the main stream PTSD, depression and drug abuse-oriented studies.


The Regional Migration-Development Nexus in Australia: What Migration? Whose Development?
Both regional resettlement of refugees, and the attraction of different kinds of migrant labor to regional areas, have been significant trends in Australia's recent migration policies. Using the concept of the migration-development nexus, we address important questions about the nature and scope of development these different policies aim to promote, and achieve. We examine the intersection of policies and initiatives implemented to encourage and support refugee settlement and regional migration in Australia with the perspectives of regionally settled migrants and refugees on their regional migration outcomes. We argue that recent government policies, and multi-stakeholder initiatives aimed at regional migration and/or settlement, cast migrants as differential contributors to regional development, useful either in terms of their skills (skilled migrants) or their labor (backpackers, seasonal workers, refugees). The co-presence of different groups of migrants in regional locations is also shaped by the fluctuating employer demands for mobile labor in combination with visa regulations. We draw on data from three projects on regional settlement, multiculturalism and mobilities to analyze three important elements of regional migration that are central to a critical analysis of the nexus between rural migration and development in regional Australia: the complex roles of employers; the embedding of regional migration in migrants' life courses; and the tension between long-term migration outcomes and quick fixes. By focusing on development as it is experienced by migrants themselves and interpreted by different stakeholders in regional migration, we draw attention to the limitations of a purely instrumental view of migrants as agents of regional development. We argue that the sustainability of regional migration policies will depend on recognizing the important role of migrants' hopes, needs and aspirations as well as their rights, and the unintended human costs and consequences of exclusively economically driven migration policy design.


Toward a situation-specific theory of breastfeeding.
This article reports a preliminary situation-specific theory of breastfeeding developed using an integrative, inductive approach (Im & Meleis, 1999). The theory purports varying levels of conflict versus congruity existing between the mother/infant dyad, a mother and her support networks, and both between and within the networks, all of which either block or facilitate breastfeeding. The theory proposes that to decrease conflict, professionals need to carefully consider their approach to promoting and supporting breastfeeding so as to respect the right of maternal decision-making and to avoid semblances of coercion or paternalism. "Salutary breastfeeding" is proposed as a new, ideal breastfeeding experience that is positive, healthy, and fulfilling, yet encompasses acknowledgement of diversity in maternal/infant dyads and situational context.


Metallothionein and brain inflammation.
Since the seminal discoveries of Bert Vallee regarding zinc and metallothioneins (MTs) more than 50 years ago, thousands of studies have been published concerning this fascinating story. One of the most active areas of research is the involvement of these proteins in the inflammatory response in general, and in neuroinflammation in particular. We describe the general aspects of the inflammatory response, highlighting the essential role of the major cytokine interleukin-6, and review briefly the expression and function of MTs in the central nervous system in the context of neuroinflammation. Particular attention is paid to the Tg2576 Alzheimer disease mouse model and the preliminary results obtained in mice into which human Zn(7)MT-2A was injected, which suggest a reversal of the behavioral deficits while enhancing amyloid plaque load and gliosis.


The inflammatory response in myocardial infarction.
One of the major therapeutic goals of modern cardiology is to design strategies aimed at minimizing myocardial necrosis and optimizing cardiac repair following myocardial infarction. However, a sound understanding of the biology is necessary before a specific intervention is pursued on a therapeutic basis. This review summarizes our current understanding of the cellular and molecular mechanisms regulating the inflammatory response following myocardial ischemia and reperfusion. Myocardial necrosis induces complement activation and free radical generation, triggering a cytokine cascade initiated by Tumor Necrosis Factor (TNF)-alpha release. If reperfusion of the infarcted area is initiated, it is attended by an intense inflammatory reaction. Interleukin (IL)-8 synthesis and C5a activation have a crucial role in recruiting neutrophils in the ischemic and reperfused myocardium. Neutrophil infiltration is regulated through a complex sequence of molecular steps involving the selectins and the integrins, which mediate leukocyte rolling and adhesion to the endothelium. Marginated neutrophils exert potent cytotoxic effects through the release of proteolytic enzymes and the adhesion with Intercellular Adhesion Molecule (ICAM)-1 expressing cardiomyocytes. Despite this potential injury, substantial evidence suggests that reperfusion enhances cardiac repair improving patient survival; this effect may be in part related to the inflammatory response. Monocyte Chemoattractant Protein (MCP)-1 is also markedly upregulated in the infarcted myocardium inducing recruitment of mononuclear cells in the injured areas. Monocyte-derived macrophages and mast cells may produce cytokines and growth factors necessary for fibroblast proliferation and neovascularization, leading to effective repair and scar formation. At this stage expression of inhibitory cytokines such as IL-10 may have a role in suppressing the acute inflammatory response and in regulating extracellular matrix metabolism. Fibroblasts in the healing scar undergo phenotypic changes expressing smooth muscle cell markers. Our previous review in this journal focused almost exclusively on reduction of the inflammatory injury. The current update is prompted by the potential therapeutic opportunity that the open vessel offers. By promoting more effective tissue repair, it may be possible to reduce the deleterious remodeling, that is the leading cause of heart failure and death. Elucidating the complex interactions and regulatory mechanisms responsible for cardiac repair may allow us to design effective inflammation-related interventions for the treatment of myocardial infarction.


Improving pharmacokinetic-pharmacodynamic modeling to investigate anti-infective chemotherapy with application to the current generation of antimalarial drugs.
Mechanism-based pharmacokinetic-pharmacodynamic (PK/PD) modelling is the standard computational technique for simulating drug treatment of infectious diseases with the potential to enhance our understanding of drug treatment outcomes, drug deployment strategies, and dosing regimens. Standard methodologies assume only a single drug is used, it acts only in its unconverted form, and that oral drugs are instantaneously absorbed across the gut wall to their site of action. For drugs with short half-lives, this absorption period accounts for a significant period of their time in the body. Treatment of infectious diseases often uses combination therapies, so we refined and substantially extended the PK/PD methodologies to incorporate (i) time lags and drug concentration profiles resulting from absorption across the gut wall and, if required, conversion to another active form; (ii) multiple drugs within a treatment combination; (iii) differing modes of action of drugs in the combination: additive, synergistic, antagonistic; (iv) drugs converted to an active metabolite with a similar mode of action. This methodology was applied to a case study of two first-line malaria treatments based on artemisinin combination therapies (ACTs, artemether-lumefantrine and artesunate-mefloquine) where the likelihood of increased artemisinin tolerance/resistance has led to speculation on their continued long-term effectiveness. We note previous estimates of artemisinin kill rate were underestimated by a factor of seven, both the unconverted and converted form of the artemisinins kill parasites and the extended PK/PD methodology produced results consistent with field observations. The simulations predict that a potentially rapid decline in ACT effectiveness is likely to occur as artemisinin resistance spreads, emphasising the importance of containing the spread of artemisinin resistance before it results in widespread drug failure. We found that PK/PD data is generally very poorly reported in the malaria literature, severely reducing its value for subsequent re-application, and we make specific recommendations to improve this situation.


Wireless magnetoelastic sensors for tracking degradation profiles of nitrodopamine-modified poly(ethylene glycol).
A critical property for tissue adhesives is a controllable degradation rate so that these adhesives do not act as barriers to wound healing. Typical degradation tests require large amount of samples, which can be tedious and expensive to perform. Additionally, current degradation tests are carried out in vitro under simulated physiological conditions and may not accurately reflect the complex environment that an adhesive would experience in vivo. As a means to develop a simple technique for testing tissue adhesive, a rapidly degrading adhesive hydrogel that mimics mussel adhesive proteins was coated onto magnetoelastic (ME) sensor strips to track the degradation of the adhesive remotely and in real time. Adhesive-coated ME sensors were submerged in phosphate buffer saline solution (pH 7.4) at body temperature (37 °C). Based on the change in the resonant amplitude, the degradation time was determined to be 22 min, which was in agreement with qualitative monitoring of the bulk adhesive hydrogel. Additionally, when the adhesive-coated ME sensor was incubated in a slightly acidic medium (pH 5.7), the degradation rate was drastically lengthened (3 hrs) as the hydrolysis of ester bonds is faster under basic conditions. Oscillatory rheological testing confirmed the formation and degradation of the adhesive. However, rheological test results did not accurately reflect the degradation rate of the adhesive hydrogel, potentially due to a slow exchange of acidic degradation products with the surrounding medium. ME sensor was demonstrated as a potential useful tool for evaluating the degradation rate of bioadhesives.


Evolutionary genetics: what is driving male mutation?
In mammals, most new mutations occur in males. But a study of the evolution of a human X to Y chromosomal translocation has revealed a sex bias much lower than previous estimates. Patterns of substitution suggest that differential methylation between male and female germ lines is a key determinant of the mutation rate.


Consortin, a trans-Golgi network cargo receptor for the plasma membrane targeting and recycling of connexins.
Targeting of numerous transmembrane proteins to the cell surface is thought to depend on their recognition by cargo receptors that interact with the adaptor machinery for anterograde traffic at the distal end of the Golgi complex. We report here on consortin, a novel integral membrane protein that is predicted to be intrinsically disordered, i.e. that contains large segments whose native state is unstructured. We identified consortin as a binding partner of connexins, the building blocks of gap junctions. Consortin is located at the trans-Golgi network (TGN), in tubulovesicular transport organelles, and at the plasma membrane. It directly interacts with the TGN clathrin adaptors GGA1 and GGA2, and disruption of this interaction by expression of a consortin mutant lacking the acidic cluster-dileucine (DXXLL) GGA interaction motif causes an intracellular accumulation of several connexins. RNA interference-mediated silencing of consortin expression in HeLa cells blocks the cell surface targeting of these connexins, which accumulate intracellularly, whereas partial depletion and redistribution of the consortin pool slows down the intracellular degradation of gap junction plaques. Altogether, our results show that, by studying connexin trafficking, we have identified the first TGN cargo receptor for the targeting of transmembrane proteins to the plasma membrane. The identification of consortin provides in addition a potential target for therapies aimed at diseases in which connexin traffic is altered, including cardiac ischemia, peripheral neuropathies, cataracts and hearing impairment. Sequence accession numbers. GenBank: Human CNST cDNA, NM_152609; mouse Cnst cDNA, NM_146105.


Should aspirin be replaced with ADP blockers and anti-GPVI to manage thrombosis?
Platelets have a pivotal role in maintaining cardiovascular homeostasis. They are kept docile by endothelial-derived mediators. Aberration in haemostatic balance predisposes an individual to an elevated risk of a prothrombotic environment. Anti-platelet therapy has been a key component to reduce this risk. However, understanding how these medications affect the balance between the activation and inhibition of platelets is critical. There is no evidence that a key anti-platelet therapy - aspirin, may not be the most efficacious medicine of choice, as it can compromise both platelet inhibition and activation pathways. In this review, the rationale of aspirin as an anti-thrombotic drug has been critically discussed. This review looks at how recently published trials are raising key questions about the efficacy and safety of aspirin in countering cardiovascular diseases. There is an increasing portfolio of evidence that identifies that although aspirin is a very cheap and accessible drug, it may be used in a manner that is not always beneficial to a patient, and a more nuanced and targeted use of aspirin may increase its clinical benefit and maximize patient response. The questions about the use of aspirin raise the potential for changes in its clinical use for dual anti-platelet therapy. This highlights the need to ensure that treatment is targeted in the most effective manner and that other anti-platelet therapies may well be more efficacious and beneficial for CVD patients in their standard and personalized approaches.


Tau Positron Emission Tomographic Findings in a Former US Football Player With Pathologically Confirmed Chronic Traumatic Encephalopathy.
Biomarkers for chronic traumatic encephalopathy (CTE) are currently lacking. The radiotracer fluorine F 18-labeled (18F)-flortaucipir (FTP) detects tau pathology in Alzheimer disease, and positron emission tomography (PET) with FTP shows elevated binding in individuals at risk for CTE. No study, however, has assessed the correlation between in vivo FTP PET and postmortem tau in CTE. To assess the regional association between in vivo FTP binding and postmortem tau pathology in a patient with pathologically confirmed CTE. A white male former National Football League player with 17 years of US football exposure was clinically diagnosed with traumatic encephalopathy syndrome at a neurology tertiary referral center. 18F-Fludeoxyglucose, carbon 11-labeled Pittsburgh compound B, and FTP PET were performed 52 months prior to death, and magnetic resonance imaging, 50 months prior to death. Brain images were assessed qualitatively for abnormalities blinded to autopsy data. Autopsy was performed using a neurodegenerative research protocol. The FTP standardized uptake value ratios (inferior cerebellar gray reference region) and W-score (age-adjusted z-score) maps were compared with phosphorylated tau immunohistochemical analysis with monoclonal antibody CP13. Qualitative and quantitative comparisons between antemortem FTP PET and tau pathology at autopsy. Flortaucipir uptake was distributed in a patchy, frontotemporal-predominant pattern that overlapped with regions showing neurodegeneration on magnetic resonance imaging and hypometabolism on 18F-fludeoxyglucose PET. Pathological assessment revealed stage 4 CTE; limbic argyrophilic grain disease; stage 2 limbic-predominant, age-related transactive response DNA-binding protein 43 encephalopathy; and Braak neurofibrillary tangle stage 3. 18F-Flortaucipir W-maps matched areas of high postmortem tau burden in left fusiform and inferior temporal gyri and juxtacortical frontal white matter. High FTP W-scores with low tau burden were found in the basal ganglia, thalamus, motor cortex, and calcarine cortex. No regions with low FTP W-scores corresponded to areas with high pathological tau burden. A modest correlation, which did not reach statistical significance (ρ = 0.35, P = .17), was found between FTP standardized uptake value ratio and tau area fraction at the regional level. In this patient, FTP PET findings during life showed a modest correspondence with postmortem pathology in CTE. These findings suggest that FTP may have limited utility as a tau biomarker in CTE.


Lymph Node Micrometastases and In-Transit Metastases from Melanoma: In Vivo Detection with Multispectral Optoacoustic Imaging in a Mouse Model.
Purpose To study whether multispectral optoacoustic tomography (MSOT) can serve as a label-free imaging modality for the detection of lymph node micrometastases and in-transit metastases from melanoma on the basis of the intrinsic contrast of melanin in comparison to fluorine 18 fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT). Materials and Methods The study was approved by the institutional animal care and use committee. Sequential MSOT was performed in a mouse B16F10 melanoma limb lymph node metastasis model (n = 13) to survey the development of macro-, micro- and in-transit metastases (metastases that are in transit from the primary tumor site to the local nodal basin) in vivo. The in vitro limit of detection was assessed in a B16F10 cell phantom. Signal specificity was determined on the basis of a simultaneous lymphadenitis (n = 4) and 4T1 breast cancer lymph metastasis (n = 2) model. MSOT was compared with intravenous FDG PET/CT. The diagnosis was assessed with histologic examination. Differences in the signal ratio (metastatic node to contralateral limb) between the two modalities were determined with the two-tailed paired t test. Results The mean signal ratios acquired with MSOT in micrometastases (2.5 ± 0.3, n = 6) and in-transit metastases (8.3 ± 5.8, n = 4) were higher than those obtained with FDG PET/CT (1.1 ± 0.5 [P < .01] and 1.3 ± 0.6 [P < .05], respectively). MSOT was able to help differentiate even small melanoma lymph node metastases from the other lymphadenopathies (P < .05 for both) in vivo, whereas FDG PET/CT could not (P > .1 for both). In vitro, the limit of detection was at an approximate cell density of five cells per microliter (P < .01). Conclusion MSOT enabled detection of melanoma lymph node micrometastases and in-transit metastases undetectable with FDG PET/CT and helped differentiate melanoma metastasis from other lymphadenopathies. (©) RSNA, 2016 Online supplemental material is available for this article.


The effect of ketamine on the separation anxiety and emergence agitation in children undergoing brief ophthalmic surgery under desflurane general anesthesia.
Emergence agitation (EA) frequently occurs after desflurane anesthesia in children. Ketamine, because of its sedative and analgesic properties, might be useful for the management of separation anxiety and EA. We investigated the preventive effect of ketamine on separation anxiety and EA after desflurane anesthesia in children for brief ophthalmic surgery. Sixty children, ranging in age from 2-8 years old, undergoing brief ophthalmic surgery were randomly allocated to one of the 3 groups: group C received normal saline, group K1.0 received ketamine 1.0 mg/kg intravenously before entering the operating room, or group K0.5 received ketamine 0.5 mg/kg 10 min before the end of the surgery. Before induction, the separation anxiety score was evaluated. Extubation time, post-anesthesia care unit stay time, postoperative nausea and vomiting, emergence agitation, and pain were assessed. The group K1.0 had a lower separation anxiety score compared with groups K0.5 and C. Extubation time in group K0.5 was significantly prolonged compared with groups K1.0 and C. The incidence of EA and the modified Children's Hospital of Eastern Ontario Pain Scale were significantly lower in group K1.0 and group K0.5 compared to group C, but there was no significant difference between groups K1.0 and K0.5. In children undergoing brief ophthalmic surgery with desflurane anesthesia, ketamine 1.0 mg/kg administered before entering the operating room reduced separation anxiety, postoperative pain, and incidence of EA without delay in recovery.


Laser trapping/confocal Raman spectroscopic characterization of PLGA-PEG nanoparticles.
We have immobilized poly(ethylene glycol) (PEG) on the surfaces of poly(lactic-co-glycolic acid) (PLGA) nanoparticles by two different chemical methods, i.e., SOCl2 halogenate-alcoholysis and DCC dehydration. The immobilized PLGA nanoparticles were characterized by DLS, 1H NMR, FT-IR and laser trapping/confocal Raman spectroscopic techniques. As a result, especially the Raman spectra which were measured after optically trapping ca. 10 individual nanoparticles in solution indicated that the PLGA nanoparticles were successfully immobilized with the PEG by the chemical methods.


A fidget spinner for the point-of-care diagnosis of urinary tract infection.
The point-of-care detection of pathogens in biological samples in resource-limited settings should be inexpensive, rapid, portable, simple and accurate. Here, we describe a custom-made fidget spinner that rapidly concentrates pathogens in 1-ml samples of undiluted urine by more than 100-fold for the on-device colorimetric detection of bacterial load and pathogen identification. In Tiruchirappalli, India, the device enabled the on-site detection of infection with the naked eye within 50 min in urine samples from 39 patients suspected of having a urinary tract infection. We also show that, in 30 clinical samples of urinary tract infection, the device can be used to perform an antimicrobial susceptibility test for the antimicrobial drugs ciprofloxacin and cefazolin within 120 min. The fidget spinner could be used in low-resource settings as an inexpensive handheld point-of-care device for the rapid concentration and detection of pathogens in urine samples.


Bee Venom: From Venom to Drug.
Insects of the order Hymenoptera have a defensive substance that contains many biologically active compounds. Specifically, venom from honeybees (Apis mellifera) contains many enzymes and peptides that are effective against various diseases. Different research papers stated the possibility of using bee venom (a direct bee sting or in an injectable form) in treating several complications; either in vivo or in vitro. Other reports used the active fractions of bee venom clinically or at labratory scale. Many reports and publications have stated that bee venom and its constituents have multiple biological activities including anti-microbial, anti-protozoan, anti-cancer, anti-inflammatory, and anti-arthritic properties. The present review aims to refer to the use of bee venom itself or its fractions in treating several diseases and counteracting drug toxicities as an alternative protocol of therapy. The updated molecular mechanisms of actions of bee venom and its components are discussed in light of the previous updated publications. The review also summarizes the potential of venom loaded on nanoparticles as a drug delivery vehicle and its molecular mechanisms. Finally, the products of bee venom available in markets are also demonstrated.


Validity of Level of Supervision Scales for Assessing Pediatric Fellows on the Common Pediatric Subspecialty Entrustable Professional Activities.
Entrustable professional activities (EPAs) represent the routine and essential activities that physicians perform in practice. Although some level of supervision scales have been proposed, they have not been validated. In this study, the investigators created level of supervision scales for EPAs common to the pediatric subspecialties and then examined their validity in a study conducted by the Subspecialty Pediatrics Investigator Network (SPIN). SPIN Steering Committee members used a modified Delphi process to develop unique scales for six of the seven common EPAs. The investigators sought validity evidence in a multisubspecialty study in which pediatric fellowship program directors and Clinical Competency Committees used the scales to evaluate fellows in fall 2014 and spring 2015. Separate scales for the six EPAs, each with five levels of progressive entrustment, were created. In both fall and spring, more than 300 fellows in each year of training from over 200 programs were assessed. In both periods and for each EPA, there was a progressive increase in entrustment levels, with second-year fellows rated higher than first-year fellows (P < .001) and third-year fellows rated higher than second-year fellows (P < .001). For each EPA, spring ratings were higher (P < .001) than those in the fall. Interrater reliability was high (Janson and Olsson's iota = 0.73). The supervision scales developed for these six common pediatric subspecialty EPAs demonstrated strong validity evidence for use in EPA-based assessment of pediatric fellows. They may also inform the development of scales in other specialties.


Nurses' perspectives of a new individualized nursing care intervention for COPD patients in primary care settings: A mixed method study.
The major challenges in Chronic Obstructive Pulmonary Disease (COPD) care are guiding a patient in daily living with the consequences of the disease, reducing the impact of symptoms and improving Health Related Quality of Life (HRQoL). The new nurse-led COPD-Guidance, Research on an Illness Perception (COPD-GRIP) intervention translates the evidence concerning illness perceptions and Health Related Quality of Life (HRQoL) into a practice nurse intervention. The aim is to explore the nurses' experiences with applying the new COPD-GRIP intervention. An explanatory mixed-method study nested in a cluster randomized trial in primary care was conducted. Pre-intervention questionnaires were sent to all participating nurses (N=24) to identify expectations. Post-intervention questionnaires identified experiences after applying the intervention followed by two focus groups to further extend exploration of findings. Questionnaires were analyzed by descriptive analyses. To identify themes the audio-taped and transcribed focus groups were independently coded by two researchers. The nurses described the intervention as a useful, structured and individualized tool to guide COPD patients in living with the consequences of COPD. Applying the intervention took less time than the nurses initially expected. The intervention enables to provide patient-centered care and to address patient needs. Barriers were encountered, especially in patients with a lower social economic status, in patients with a lower health literacy and in patients with other cultural backgrounds than the Dutch background. Nurses perceived the COPD-GRIP intervention as a feasible, individualized tool. According to the nurses, the intervention is a valuable improvement in the care for COPD patients.


Sex ratio at birth and racial differences: why do black women give birth to more females than non-black women?
The two important questions that this paper will attempt to answer are: (1) why is it that regardless of race/ethnicity or geographic location, the sex ratio data at birth show more males than females?; and (2) Why is it that regardless of geographic location compared to other racial/ethnic groups, Black women or Women of sub-Saharan Black African descent tend to give birth to more females? Or to put this question the other way around, compared to Black women, why do non-Black women give birth to more males?


Longitudinal patterns of breast cancer screening: mammography, clinical, and breast self-examinations in a rural and urban setting.
We identified breast screening patterns over time and patterns among women residing in rural and urban areas by sociodemographic factors. This study employs a longitudinal design over 9 years from 2001 on 11,200 women aged 50 to 55 from the Australian Longitudinal Study on Women's Health. Area of residence was defined in accordance with the accessibility remoteness index of Australia Plus. Breast screening measures included mammography utilization, clinical breast examinations (CBE), and breast self-examinations (BSE). Most women had a mammogram in the past 2 years in combination with CBE or BSE or both. Despite poorer access to mammography services, women residing in rural areas had similar mammography screening rates to their urban counterparts. Women residing in rural areas were less likely to have CBEs, but more likely to conduct BSEs. The breast screening behaviors were generally consistent over time. The poorer breast cancer survival among rural women is unlikely to be explained by differences in mammography service use. A substantial proportion of the population may be experiencing overscreening by conducting all three types of breast screening.


'We keep it secret so no one should know'--a qualitative study to explore young schoolgirls attitudes and experiences with menstruation in rural western Kenya.
Keeping girls in school offers them protection against early marriage, teen pregnancy, and sexual harms, and enhances social and economic equity. Studies report menstruation exacerbates school-drop out and poor attendance, although evidence is sparse. This study qualitatively examines the menstrual experiences of young adolescent schoolgirls. The study was conducted in Siaya County in rural western Kenya. A sample of 120 girls aged 14-16 years took part in 11 focus group discussions, which were analysed thematically. The data gathered were supplemented by information from six FGDs with parents and community members. Emergent themes were: lack of preparation for menarche; maturation and sexual vulnerability; menstruation as an illness; secrecy, fear and shame of leaking; coping with inadequate alternatives; paying for pads with sex; and problems with menstrual hygiene. Girls were unprepared and demonstrated poor reproductive knowledge, but devised practical methods to cope with menstrual difficulties, often alone. Parental and school support of menstrual needs is limited, and information sparse or inaccurate. Girls' physical changes prompt boys and adults to target and brand girls as ripe for sexual activity including coercion and marriage. Girls admitted 'others' rather than themselves were absent from school during menstruation, due to physical symptoms or inadequate sanitary protection. They described difficulties engaging in class, due to fear of smelling and leakage, and subsequent teasing. Sanitary pads were valued but resource and time constraints result in prolonged use causing chafing. Improvised alternatives, including rags and grass, were prone to leak, caused soreness, and were perceived as harmful. Girls reported 'other girls' but not themselves participated in transactional sex to buy pads, and received pads from boyfriends. In the absence of parental and school support, girls cope, sometimes alone, with menarche in practical and sometimes hazardous ways. Emotional and physical support mechanisms need to be included within a package of measures to enable adolescent girls to reach their potential.


Spoiled breast milk and bad water; local understandings of diarrhea causes and prevention in rural Sierra Leone.
Globally, diarrhea remains a leading killer of young children. In Sierra Leone, one in seven children die before their fifth birthday and diarrhea is a leading cause. Studies that emphasize the demand-side of health interventions -- how caregivers understand causation and prevention of diarrhea -- have been neglected in research and programming. We undertook applied qualitative research including 68 in-depth interviews and 36 focus group discussions with mothers, fathers and older female caretakers to examine the causes and prevention of childhood diarrhea in villages near and far from health facilities across four rural districts. Verbal consent was obtained. Respondents reported multiple, co-existing descriptions of causation including: contaminated water and difficulties accessing clean water; exposure to an unclean environment and poor food hygiene; contaminated breast milk due to sexual intercourse, overheated breast milk or bodily maternal conditions such as menstruation or pregnancy; and dietary imbalances and curses. Respondents rarely discussed the role of open defecation or the importance of handwashing with soap in preventing diarrhea. Categorizing behaviors as beneficial, harmful, non-existent or benign enables tailored programmatic recommendations. For example, respondents recognized the value of clean water and we correspondingly recommend interventions that reinforce consumption of and access to clean water. Second, respondents report denying "contaminated" breast milk to breastfeeding children. This is a harmful practice that merits attention. Third, the role of open defecation and poor hygiene in causing diarrhea is less understood and warrants introduction or clarification. Finally, the role of exposed feet or curses in causing diarrhea is relatively benign and does not necessitate programmatic attention. Further research supportive of communication and social mobilization strategies building on these findings is required to ensure that improved understanding regarding diarrhea causation translates into improved diarrhea prevention.


Parasitic versus nutritional regulation of natural fish populations.
Although parasites are expected to affect their host's fitness, quantitative proof for impacts of parasitism on wild populations is hampered by confounding environmental factors, including dietary resource. Herein, we evaluate whether the physiological conditions of European perch (Perca fluviatilis) in three large peri-alpine lakes (Geneva, Annecy, and Bourget) depend on (a) the nutritional status of the juvenile fish, as revealed by stable isotope and fatty acid compositions, (b) the prevalence of the tapeworm Triaenophorus nodulosus, a parasite transmitted to perch through copepod preys, or (c) interactive effects of both factors. At the scale of lake populations, the deficit in growth and fat storage of juvenile perch during their first summer coincides with a high parasite prevalence and also a low quality of dietary resource. Yet, at the individual level, parasites had no evident effect on the growth of the juvenile perch, while impacts on fat storage appeared only at the highest prevalence of the most infected lake. Fatty acid and stable isotope analyses of fish tissue do not reveal any impact of T. nodulosus on diet, physiology, and feeding behaviour of fish within lakes. Overall, we found a low impact of parasitism on the physiological condition and trophic status of juvenile perch at the end of their first summer. We find instead that juvenile perch growth and fat storage, both factors tied to their winter survival, are under strong nutritional constraints. However, the coinciding nutritional constraints and parasite prevalence of perch juveniles in these three lakes may result from the indirect effect of lake nutrient concentrations, which, as a major control of zooplankton communities, simultaneously regulate both the dietary quality of fish prey and the host-parasite encounter rates.


HIV risk reduction intervention among traditionally circumcised young men in South Africa: a cluster randomized control trial.
The aim of this study was to test a 180-minute group HIV risk-reduction counseling intervention trial with men undergoing traditional circumcision in South Africa to reduce behavioral disinhibition (false security) as a result of the procedure. A cluster randomized controlled trial design was employed using a sample of 160 men, 80 in the experimental group and 80 in the control group. Comparisons between baseline and 3-month follow-up assessments on key behavioral outcomes were completed. We found that behavioral intentions, risk-reduction skills, and male role norms did not change in the experimental compared to the control condition. However, HIV-related stigma beliefs were significantly reduced in both conditions over time. These findings show that one small-group HIV risk-reduction intervention did not reduce sexual risk behaviors in recently traditionally circumcised men at high risk for behavioral disinhibition.


Angiogenesis during primate placentation in health and disease.
Normal embryonic development is dependent upon a sufficient oxygen, nutrient and waste exchange through the placenta. In primates including humans, this exchange is attained by successful haemochorial placentation which requires the transformation of maternal intramyometrial spiral arterioles by trophoblast invasion to gain uteroplacental circulation, and establishment and maintenance of a competent fetoplacental vasculature. Thus, trophoblast and endothelial cell differentiation, proliferation and invasion occurring during placentation have to be tightly regulated. This review focuses on the diverse developmental steps during haemochorial placentation in humans and other primates and the possible involvement of angiogenic growth factors (vascular endothelial growth factor (VEGF) and angiopoietins (Ang)) in these processes, highlighting the importance of specific actions of angiogenic ligand-receptor pairs. It is hypothesized that VEGF/VEGF-R1 and Ang-1/Tie receptor 2 (Tie-2) may regulate trophoblast differentiation and invasion; VEGF/VEGF-R2 and Ang-1/Tie-2 may promote fetoplacental vascular development and stabilization; and Ang-2/Tie-2 may be involved in maternal vascular remodelling. The importance of a tight regulation of angiogenic factors and their endogenous antagonists for normal development of the placenta is demonstrated by failure of this system, resulting in abnormal placenta vascularization and trophoblast invasion associated with intrauterine growth retardation or pre-eclampsia.


Comparative plant development: the time of the leaf?
A key problem in developmental biology is understanding the origin of morphological innovations. Comparative studies in plants with different leaf morphologies indicate that the developmental pathway defined by KNOTTED1-type homeodomain proteins could be involved in generating different leaf forms. The differential expression of regulatory proteins has emerged as an important factor in driving morphological innovations in the plant kingdom--an idea that is well supported by quantitative trait locus analyses.


Heat shock protein and gliadin peptide promote development of peptidase antibodies in children with autism and patients with autoimmune disease.
Searching for a mechanism underlying autoimmunity in autism, we postulated that gliadin peptides, heat shock protein 60 (HSP-60), and streptokinase (SK) bind to different peptidases resulting in autoantibody production against these components. We assessed this hypothesis in patients with autism and in those with mixed connective tissue diseases. Associated with antigliadin and anti-HSP antibodies, children with autism and patients with autoimmune disease developed anti-dipeptidylpeptidase I (DPP I), anti-dipeptidylpeptidase IV (DPP IV [or CD26]) and anti-aminopeptidase N (CD13) autoantibodies. A significant percentage of autoimmune and autistic sera were associated with elevated immunoglobulin G (IgG), IgM, or IgA antibodies against three peptidases, gliadin, and HSP-60. These antibodies are specific, since immune absorption demonstrated that only specific antigens (e.g., DPP IV absorption of anti-DPP IV), significantly reduced IgG, IgM, and IgA antibody levels. For direct demonstration of SK, HSP-60, and gliadin peptide binding to DPP IV, microtiter wells coated with DPP IV were reacted with SK, HSP-60, and gliadin. They were then reacted with anti-DPP IV or anti-SK, anti-HSP, and antigliadin antibodies. Adding SK, HSP-60, and gliadin peptides to DPP IV resulted in 27 to 43% inhibition of the DPP IV-anti-DPP IV reaction, but DPP IV-positive peptides caused 18 to 20% enhancement of antigen-antibody reactions. We propose that (i) superantigens (e.g., SK and HSP-60) and dietary proteins (e.g., gliadin peptides) in individuals with predisposing HLA molecules bind to aminopeptidases and (ii) they induce autoantibodies to peptides and tissue antigens. Dysfunctional membrane peptidases and autoantibody production may result in neuroimmune dysregulation and autoimmunity.


Effects of different insulin administrations on plasma amino acid profile in insulin-dependent diabetic patients.
It has long been recognized that diabetes mellitus is characterized by alterations in amino acid metabolism. Our aim was to evaluate the fasting amino acid concentrations in insulin-dependent diabetic subjects treated with different kinds of insulin therapy and with respect to the resulting different metabolic control. Four groups of patients were studied: Group 1: eight type 1 diabetic patients with two daily insulin injections; Group 2: eight type 1 diabetic patients with continuous subcutaneous insulin infusion; Group 3: five type 1 diabetic patients with continuous intraperitoneal insulin infusion; Group 4: six patients with total pancreatectomy. In all patients fasting amino acid profile was determined. Group 1 was also studied after 24 hr insulin withdrawal and after acute normalization of glycaemia. Our data demonstrated that the patients with a mild hyperglycaemia showed a significant increase in plasma amino acid levels. The continuous subcutaneous or intraperitoneal insulin infusion normalized amino acid profile at the expense of peripheral hyperinsulinemia, suggesting the presence of insulin resistance with regard of amino acid metabolism. The acute normoglycaemia demonstrated that plasma branched chain amino acid levels are more sensitive than plasma glucose to insulin action. Moreover, the amino acid pattern of euglycaemic total pancreatectomized patients suggested that a basal level of glucagon is essential in regulating glycogenic amino acid plasma concentrations.


Standardised clinical data from patients with primary ciliary dyskinesia: FOLLOW-PCD.
Clinical data on primary ciliary dyskinesia (PCD) are limited, heterogeneous and mostly derived from retrospective chart reviews, leading to missing data and unreliable symptoms and results of physical examinations. We need standardised prospective data collection to study phenotypes, severity and prognosis and improve standards of care. A large, international and multidisciplinary group of PCD experts developed FOLLOW-PCD, a standardised clinical PCD form and patient questionnaire. We identified existing forms for clinical data collection via the Better Experimental Approaches to Treat PCD (BEAT-PCD) COST Action network and a literature review. We selected and revised the content items with the working group and patient representatives. We then revised several drafts in an adapted Delphi process, refining the content and structure. FOLLOW-PCD has a modular structure, to allow flexible use based on local practice and research focus. It includes patient-completed versions for the modules on symptoms and lifestyle. The form allows a comprehensive standardised clinical assessment at baseline and for annual reviews and a short documentation for routine follow-up. It can either be completed using printable paper forms or using an online REDCap database. Data collected in FOLLOW-PCD version 1.0 is available in real-time for national and international monitoring and research. The form will be adapted in the future after extensive piloting in different settings and we encourage the translation of the patient questionnaires to multiple languages. FOLLOW-PCD will facilitate quality research based on prospective standardised data from routine care, which can be pooled between centres, to provide first-line and real-time evidence for clinical decision-making.


NCCN Guidelines Insights: Chronic Myeloid Leukemia, Version 1.2017.
The NCCN Guidelines for Chronic Myeloid Leukemia (CML) provide recommendations for the management of chronic-phase and advanced-phase CML in adult patients. The median age of disease onset is 67 years. However, because CML occurs in all age groups, clinical care teams should be prepared to address issues relating to fertility and pregnancy with patients who are of reproductive age at the time of diagnosis. CML is relatively rare in children and there are no evidence-based recommendations for the management of CML in pediatric population. These NCCN Guidelines Insights discuss special considerations for the management of CML during pregnancy and for the management of CML in the pediatric population.


The success of various management techniques used in South African children with type 1 diabetes mellitus.
Despite the availability of international guidelines for the treatment of type 1 diabetes mellitus in children, important aspects of treatment are not accessible to all young patients in South Africa (SA). To investigate factors in diabetes management strategies that are associated with poor glycaemic control and decreased quality of life (QoL) in SA children with type 1 diabetes mellitus (T1DM). Eighty children (mean (standard deviation) age 12.9 (2.7) years) with T1DM were asked to answer standardised questionnaires on demographics, management techniques used and perceptions of diabetes. The height and weight of each child was recorded and glycosylated haemoglobin (HbA1c) measured. Informed consent and assent for each participant was obtained before enrolment. A total of 51.4% of the participants had poor metabolic control, with an HbA1c level >10.0% (86 mmol/mol). Factors in clinical practice found to have a significant association with decreased HbA1c and/or QoL were healthcare system (p<0.001), insulin administration (p=0.001), correction dose (p=0.002), carbohydrate counting (p<0.001) and number of severe hyperglycaemic events (p=0.048). Regular exercise did not show any association with HbA1c classification or QoL. Children from single-parent households were prone to unsuccessful diabetes management regardless of treatment techniques used (p=0.002). The use of premixed insulin without access to rapid-acting insulin, absence of correction doses for hyperglycaemia and lack of carbohydrate counting showed significant association with poor diabetes management. Some recommendations regarding the adoption of more effective diabetes management strategies in the public healthcare system are suggested.


Number sense in human infants.
Four experiments used a preferential looking method to investigate 6-month-old infants' capacity to represent numerosity in visual-spatial displays. Building on previous findings that such infants discriminate between arrays of eight versus 16 discs, but not eight versus 12 discs (Xu & Spelke, 2000), Experiments 1 and 2 investigated whether infants' numerosity discrimination depends on the ratio of the two set sizes with even larger numerosities. Infants successfully discriminated between arrays of 16 versus 32 discs, but not 16 versus 24 discs, providing evidence that their discrimination shows the set-size ratio signature of numerosity discrimination in human adults, children and many non-human animals. Experiments 3 and 4 addressed a controversy concerning infants' ability to discriminate large numerosities (observed under conditions that control for total filled area, array size and density, item size and correlated properties such as brightness: Brannon, 2002; Xu, 2003b; Xu & Spelke, 2000) versus small numerosities (not observed under conditions that control for total contour length: Clearfield & Mix, 1999). To investigate the sources of these differing findings, Experiment 3 tested infants' large-number discrimination with controls for contour length, and Experiment 4 tested small-number discrimination with controls for total filled area. Infants successfully discriminated the large-number displays but showed no evidence of discriminating the small-number displays. These findings provide evidence that infants have robust abilities to represent large numerosities. In contrast, infants may fail to represent small numerosities in visual-spatial arrays with continuous quantity controls, consistent with the thesis that separate systems serve to represent large versus small numerosities.


A new strategy for gene targeting and functional proteomics using the DT40 cell line.
DT40 cells derived from chicken B lymphocytes exhibit exceptionally high homologous recombination rates. Therefore, they can be used as a convenient tool and model for gene targeting experiments. However, lack of efficient cloning strategies, protein purification protocols and a well annotated protein database limits the utility of these cells for proteomic studies. Here we describe a fast and inexpensive experimental pipeline for protein localization, quantification and mass spectrometry-based interaction studies using DT40 cells. Our newly designed set of pQuant vectors and a sequence- and ligation-independent cloning (SLIC) strategy allow for simple and efficient generation of gene targeting constructs, facilitating homologous-recombination-based protein tagging on a multi-gene scale. We also report proof of principle results using the key proteins involved in RNA decay, namely EXOSC8, EXOSC9, CNOT7 and UPF1.


State-of-the-art computation of the rotational and IR spectra of the methyl-cyclopropyl cation: hints on its detection in space.
Recent measurements by the Cassini Ion Neutral Mass Spectrometer demonstrated the presence of numerous carbocations in Titan's upper atmosphere. In [Ali et al., Planet. Space Sci., 2013, 87, 96], an analysis of these measurements revealed the formation of the three-membered cyclopropenyl cation and its methyl derivatives. As a starting point of a future coordinated effort of laboratory experiments, quantum-chemical calculations, and astronomical observations, in the present work the molecular structure and spectroscopic properties of the methyl-cyclopropenyl cation have been investigated by means of state-of-the-art computational approaches in order to simulate its rotational and infrared spectra. Rotational parameters have been predicted with an expected accuracy better than 0.1% for rotational constants and on the order of 1-2% for centrifugal-distortion terms. As for the infrared spectrum, despite the challenge of a large amplitude motion, fundamental transitions have been computed to a good accuracy, i.e., the uncertainties are expected to be smaller than 5-10 wavenumbers.


Improved outcome at end of treatment in the collaborative Wilms tumour Africa project.
The Collaborative Wilms Tumour (WT) Africa Project has implemented an adapted WT treatment guideline in sub-Saharan Africa as a multi-centre prospective clinical trial. A retrospective, baseline evaluation of end-of-treatment outcome was performed for a 2-year period prior to the introduction of this guideline. The collaborative project aims to reduce both treatment abandonment and death during treatment to less than 10% for improving survival. All participating centres obtained local Institutional Research Board (IRB) approval and implemented the adapted WT treatment guideline. End-of-treatment outcome was documented for 2 years. It was divided into alive without evidence of disease, treatment abandonment, death during treatment and persistent disease. The outcome of children enroled in the first 2 years of the prospective clinical trial has been compared to the outcome before the start of the project. One hundred twenty-two patients were included in the baseline evaluation (2011-2012) and 133 in the first 2 years of the collaborative clinical trial (2014-2015). The percentage of patients alive without evidence of disease at the end of treatment increased from 52% (63/122) to 68% (90/133; P = 0.01). Treatment abandonment decreased from 23% (28/122) to 13% (17/133; P = 0.03). Death during treatment decreased from 21% (26/122) to 13% (17/133; P = 0.07). This collaboration, using relatively simple and low-cost interventions, led to a significant decrease in treatment abandonment and increase in survival without evidence of disease at the end of treatment.


Comparison of electrical nerve stimulation, electrical muscle stimulation and magnetic nerve stimulation to assess the neuromuscular function of the plantar flexor muscles.
As it might lead to less discomfort, magnetic nerve stimulation (MNS) is increasingly used as an alternative to electrical stimulation methods. Yet, MNS and electrical nerve stimulation (ENS) and electrical muscle stimulation (EMS) have not been formally compared for the evaluation of plantar flexor neuromuscular function. We quantified plantar flexor neuromuscular function with ENS, EMS and MNS in 10 volunteers in fresh and fatigued muscles. Central alterations were assessed through changes in voluntary activation level (VAL) and peripheral function through changes in M-wave, twitch and doublet (PS100) amplitudes. Discomfort associated with 100-Hz paired stimuli delivered with each method was evaluated on a 10-cm visual analog scale. VAL, agonist and antagonist M-wave amplitudes and PS100 were similar between the different methods in both fresh and fatigued states. Potentiated peak twitch was lower in EMS compared to ENS, whereas no difference was found between ENS and MNS for any parameter. Discomfort associated with MNS (1.5 ± 1.4 cm) was significantly less compared to ENS (5.5 ± 1.9 cm) and EMS (4.2 ± 2.6 cm) (p < 0.05). When PS100 is used to evaluate neuromuscular properties, MNS, EMS and ENS can be used interchangeably for plantar flexor neuromuscular function assessment as they provide similar evaluation of central and peripheral factors in unfatigued and fatigued states. Importantly, electrical current spread to antagonist muscles was similar between the three methods while discomfort from MNS was much less compared to ENS and EMS. MNS may be potentially employed to assess neuromuscular function of plantar flexor muscles in fragile populations.


Remission induction with L-asparaginase, vincristine, and prednisone in children with acute nonlymphoblastic leukemia.
Forty-five children with acute nonlymphoblastic leukemia in relapse received a total of 56 courses of L-asparaginase combined with vincristine and prednisone. The complete remission rate of 40% (12 of 30 trials) in patients resistant to vincristine and prednisone was almost identical to that in children still sensitive to vincristine and prednisone (42%, 11 of 26 trials). The complete remission rate of 38% (14 of 37 exposures) in those children who had not received L-asparaginase previously compared favorably with the complete remission rate in those children who had received prior L-asparaginase (47%, 9 of 19 exposures). Forty-seven of the 56 induction trials were in children with 1 or more remissions and 14 of these were in children with 3 or more prior remissions. Toxicity was minimal.


Intramolecular double proton transfer from 2-hydroxy-2-iminoacetic acid to 2-amino-2-oxoacetic acid.
The double intramolecular proton transfer process which transforms 2-hydroxy-2-iminoacetic acid into 2-amino-2-oxoacetic acid is analyzed. The MP2 and DFT calculations were performed for different tautomers and conformers of these species. Pople style (6-311++G(d,p), 6-311++G(3df,3pd)) and Dunning type (aug-cc-pVTZ) basis sets were applied. O-H...O, N-H...O, and O-H...N intramolecular hydrogen bonds were also analyzed with the use of Quantum Theory of "Atoms in Molecules" (QTAIM) as well as Natural Bond Orbitals (NBO) method. Different criteria of the hydrogen bonding existence were applied here for intramolecular interactions. It was found that some of N-H...O hydrogen bonds may be treated as blue-shifting ones.


[Ligament lesions of the knee joint in childhood].
As a matter of principle, all lesions of the knee associated with haemarthrosis must be examined for internal injuries. Ligament ruptures are rare; usually, there are bony ligament tears that show up on x-ray film. Examination of ligament stability in children is useful only if the x-ray is negative and after the knee joint has been punctured under anaesthesia, and can be justified only in that case. Diagnosis is possible only by means of x-ray films taken in forced extreme joint position in two planes, with lateral comparison. Treatment of all non-dislocated ligament tears is conservative; slight dislocations can be tolerated in cruciate and capsular ligaments, since the epiphyseal cartilage is not affected. Surgery should be performed only if tears are considerably dislocated. Proximal tears of collateral ligaments are epiphyseal fractures that can be treated conservatively only if there is no dislocation; if they are dislocated, surgery is absolutely mandatory.


Fluorescent Protein Based FRET Pairs with Improved Dynamic Range for Fluorescence Lifetime Measurements.
Fluorescence Resonance Energy Transfer (FRET) using fluorescent protein variants is widely used to study biochemical processes in living cells. FRET detection by fluorescence lifetime measurements is the most direct and robust method to measure FRET. The traditional cyan-yellow fluorescent protein based FRET pairs are getting replaced by green-red fluorescent protein variants. The green-red pair enables excitation at a longer wavelength which reduces cellular autofluorescence and phototoxicity while monitoring FRET. Despite the advances in FRET based sensors, the low FRET efficiency and dynamic range still complicates their use in cell biology and high throughput screening. In this paper, we utilized the higher lifetime of NowGFP and screened red fluorescent protein variants to develop FRET pairs with high dynamic range and FRET efficiency. The FRET variations were analyzed by proteolytic activity and detected by steady-state and time-resolved measurements. Based on the results, NowGFP-tdTomato and NowGFP-mRuby2 have shown high potentials as FRET pairs with large fluorescence lifetime dynamic range. The in vitro measurements revealed that the NowGFP-tdTomato has the highest Förster radius for any fluorescent protein based FRET pairs yet used in biological studies. The developed FRET pairs will be useful for designing FRET based sensors and studies employing Fluorescence Lifetime Imaging Microscopy (FLIM).


Inverse log-linear relationship between thyroid-stimulating hormone and free thyroxine measured by direct analog immunoassay and tandem mass spectrometry.
Accurate measurement of free thyroxine (FT(4)) is important for diagnosing and managing thyroid disorders. Most laboratories measure FT(4) by direct analogue immunoassay methods. The validity of these methods have recently been questioned. The inverse log-linear relationship between FT(4) and thyroid-stimulating hormone (TSH) is well described and provides a physiological rationale on which to base an evaluation of FT(4) assays. The study included 109 participants for whom FT(4) measurement was requested by their clinician. Samples were selected for inclusion to reflect a wide spectrum of TSH and albumin results. FT(4) and TSH were measured by use of the Siemens Immulite immunoassay (IA). FT(4) was also measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (MS-FT(4)). The inverse log-linear correlation coefficient between TSH and FT(4) was significantly better (P < 0.0001) for MS-FT(4) (0.84, 95% CI, 0.77-0.88) than for IA-FT(4) (0.45, 95% CI, 0.29-0.59). IA-FT(4) showed a significant correlation with albumin (Spearman correlation coefficient 0.45, 95% CI, 0.29-0.5, P < 0.0001) and thyroxine-binding globulin (TBG) (Spearman correlation coefficient 0.23, 95% CI, 0.05-0.41, P = 0.02). In contrast, FT(4) measurement by LC-MS/MS did not show a significant correlation with albumin or TBG. The inverse log-linear relationship between FT(4) and TSH was significantly better for FT(4) measured by LC-MS/MS than by IA. The MS-FT(4) method therefore provides FT(4) results that agree clinically with those obtained for TSH. Additionally, the significant correlation between IA-FT(4) with albumin and TBG suggests that this FT(4) method depends on binding protein concentrations and consequently does not accurately reflect FT(4).


Predictors of mask-wearing during the advent of the COVID-19 pandemic: Evidence from South Africa.
In the absence of a vaccine, the global spread of COVID-19 during 2020 has necessitated non-pharmaceutical interventions to curb the rise of cases. The article uses the health belief model and a novel rapid mobile survey to examine correlates of reported mask-wearing as a non-pharmaceutical intervention in South Africa between May and August 2020. Two-way tabulations and multivariable analysis via logistic regression modeling describe correlations between reported mask-wearing and factors of interest among a sample of 7074 adults in a two-period national longitudinal survey, the National Income Dynamics Study-Coronavirus Rapid Mobile Survey (NIDS-CRAM). In line with the health belief model, results showed that self-efficacy, the prevalence of others' mask-wearing in the same district, and affluence were positively associated with reported mask-wearing. Those who reported staying at home were significantly less likely to report wearing a mask. There was little evidence that the expected severity of the disease if contracted, affects these decisions. Hypertension, obesity, or being overweight (measured three years earlier) did not have a significant association with mask-wearing. The prevalence of mask-wearing increased significantly from May to August 2020 as COVID-19 cases increased and lockdown restrictions were eased. Contrary to the health belief model, we found that despite having a higher mortality risk, the elderly had significantly lower odds of mask-wearing. In South Africa, the mask-wearing adherence has increased rapidly. It is concerning that the elderly had lower odds of mask-wearing. This should be examined further in future research.


Bordetella Dermonecrotic Toxin Is a Neurotropic Virulence Factor That Uses CaV3.1 as the Cell Surface Receptor.
Dermonecrotic toxin (DNT) is one of the representative toxins produced by Bordetella pertussis, but its role in pertussis, B. pertussis infection, remains unknown. In this study, we identified the T-type voltage-gated Ca2+ channel CaV3.1 as the DNT receptor by CRISPR-Cas9-based genome-wide screening. As CaV3.1 is highly expressed in the nervous system, the neurotoxicity of DNT was examined. DNT affected cultured neural cells and caused flaccid paralysis in mice after intracerebral injection. No neurological symptoms were observed by intracerebral injection with the other major virulence factors of the organisms, pertussis toxin and adenylate cyclase toxin. These results indicate that DNT has aspects of the neurotropic virulence factor of B. pertussis The possibility of the involvement of DNT in encephalopathy, which is a complication of pertussis, is also discussed.IMPORTANCEBordetella pertussis, which causes pertussis, a contagious respiratory disease, produces three major protein toxins, pertussis toxin, adenylate cyclase toxin, and dermonecrotic toxin (DNT), for which molecular actions have been elucidated. The former two toxins are known to be involved in the emergence of some clinical symptoms and/or contribute to the establishment of bacterial infection. In contrast, the role of DNT in pertussis remains unclear. Our study shows that DNT affects neural cells through specific binding to the T-type voltage-gated Ca2+ channel that is highly expressed in the central nervous system and leads to neurological disorders in mice after intracerebral injection. These data raise the possibility of DNT as an etiological agent for pertussis encephalopathy, a severe complication of B. pertussis infection.


The 8-Pyrrole-Benzothiazinones Are Noncovalent Inhibitors of DprE1 from Mycobacterium tuberculosis.
8-Nitro-benzothiazinones (BTZs), such as BTZ043 and PBTZ169, inhibit decaprenylphosphoryl-β-d-ribose 2'-oxidase (DprE1) and display nanomolar bactericidal activity against Mycobacterium tuberculosis in vitro. Structure-activity relationship (SAR) studies revealed the 8-nitro group of the BTZ scaffold to be crucial for the mechanism of action, which involves formation of a semimercaptal bond with Cys387 in the active site of DprE1. To date, substitution of the 8-nitro group has led to extensive loss of antimycobacterial activity. Here, we report the synthesis and characterization of the pyrrole-benzothiazinones PyrBTZ01 and PyrBTZ02, non-nitro-benzothiazinones that retain significant antimycobacterial activity, with MICs of 0.16 μg/ml against M. tuberculosis. These compounds inhibit DprE1 with 50% inhibitory concentration (IC50) values of <8 μM and present favorable in vitro absorption-distribution-metabolism-excretion/toxicity (ADME/T) and in vivo pharmacokinetic profiles. The most promising compound, PyrBTZ01, did not show efficacy in a mouse model of acute tuberculosis, suggesting that BTZ-mediated killing through DprE1 inhibition requires a combination of both covalent bond formation and compound potency.


The diagnostic value of adiponectin multimers in healthy men undergoing screening for prostate cancer.
Adiponectin has been reported to have a prohibitory effect on prostate cancer. The goal of this study was to evaluate the diagnostic value of adiponectin multimers for prostate cancer. Total adiponectin, high- and low-molecular-weight (HMW, LMW), ratios of these measures, and body mass index (BMI) were compared in a prospective prostate cancer-screened cohort. Multivariable logistic regression was used to assess the association between adiponectin measures, their interaction with BMI, and risk of prostate cancer and Gleason score upgrading from biopsy to prostatectomy. A total of 228 prostate cancer cases and 239 controls were analyzed: 72 (31.6%) of the cancer cases were high grade (Gleason grade ≥7). Only percent HMW had a statistically significant relationship with prostate cancer (P = 0.04). Among normal and overweight men, the risk of prostate cancer increased as percent HMW increased [OR = 1.24 for a doubling of percent HMW, 95% confidence interval (CI), 0.41-3.75 and OR = 1.81; 95% CI, 1.02-3.20, respectively], whereas among obese men, the risk of prostate cancer decreased (OR = 0.62; 95% CI, 0.32-1.18). Among 97 patients who underwent radical prostatectomy, there was no association between Gleason score upgrading and any of the adiponectin multimers. This study was unable to confirm the utility of total adiponectin as a biomarker for prostate cancer risk. For the adiponectin multimers, only HMW showed increases with prostate cancer but not in all weight classes. Although adiponectin may play a role in the pathogenesis of prostate cancer, our results do not support adiponectin multimers as biomarkers of detection.


The essentiality of arachidonic acid and docosahexaenoic acid.
The purpose of this review is to correlate the clinical finding that patients receiving parenteral nutrition with a fish oil-based lipid emulsion do not develop essential fatty acid deficiency (EFAD) with an experimental murine model, thus showing that arachidonic acid (AA) and docosahexaenoic acid (DHA) are likely to be the essential fatty acids. Conventional belief is that linoleic acid (LA, omega-6) and alpha-linolenic acid (ALA, omega-3) are the essential fatty acids (EFAs). We have shown that a fish oil-based lipid emulsion containing AA (omega-6) and docosahexaenoic acid (omega-3) and insignificant quantities of LA and ALA is efficacious in the treatment of parenteral nutrition-associated liver disease (PNALD), a major cause of liver-related morbidity and mortality. The prospect of using a fish oil-based lipid emulsion as monotherapy has raised concerns of EFAD development, hindering its adoption into clinical practice. Data from patients in our institution who received PN with a fish oil-based lipid emulsion was reviewed for clinical and biochemical evidence of EFAD, defined as an elevated triene-tetraene ratio (Mead acid/AA>0.2). We also investigated the minimum amount of fish oil required to prevent EFAD in a murine model and determined whether DHA and AA alone can prevent EFAD. No patients receiving PN with a fish oil-based lipid emulsion in our institution have developed biochemical or clinical evidence of EFAD such as an elevated triene-tetraene ratio, growth retardation or dermatitis. This observation parallels our previously published animal studies, which demonstrated prevention of EFAD when 13% of total calories were from fish oil. Moreover, current work in our laboratory shows that AA and DHA provision alone is sufficient to prevent biochemical and physiologic evidence of EFAD in a murine model. When dosed appropriately, fish oil-based lipid emulsions contain sufficient EFAs to prevent EFAD. Furthermore, AA and DHA alone may be the true EFAs.


Differentiation of monkey embryonic stem cells into neural lineages.
Embryonic stem (ES) cells are self-renewing, pluripotent, and capable of differentiating into all of the cell types found in the adult body. Therefore, they have the potential to replace degenerated or damaged cells, including those in the central nervous system. For ES cell-based therapy to become a clinical reality, translational research involving nonhuman primates is essential. Here, we report monkey ES cell differentiation into embryoid bodies (EBs), neural progenitor cells (NPCs), and committed neural phenotypes. The ES cells were aggregated in hanging drops to form EBs. The EBs were then plated onto adhesive surfaces in a serum-free medium to form NPCs and expanded in serum-free medium containing fibroblast growth factor (FGF)-2 before neural differentiation was induced. Cells were characterized at each step by immunocytochemistry for the presence of specific markers. The majority of cells in complex/cystic EBs expressed antigens (alpha-fetal protein, cardiac troponin I, and vimentin) representative of all three embryonic germ layers. Greater than 70% of the expanded cell populations expressed antigenic markers (nestin and musashi1) for NPCs. After removal of FGF-2, approximately 70% of the NPCs differentiated into neuronal phenotypes expressing either microtubule-associated protein-2C (MAP2C) or neuronal nuclear antigen (NeuN), and approximately 28% differentiated into glial cell types expressing glial fibrillary acidic protein. Small populations of MAP2C/NeuN-positive cells also expressed tyrosine hydroxylase (approximately 4%) or choline acetyltransferase (approximately 13%). These results suggest that monkey ES cells spontaneously differentiate into cells of all three germ layers, can be induced and maintained as NPCs, and can be further differentiated into committed neural lineages, including putative neurons and glial cells.


[The surgery of pharyngoesophageal carcinoma with vascular anastomosis].
Reestablishment of gastrointestinal continuity following resection of pharyngoesophageal carcinoma can be a challenging problem. Recent developments in vascular surgery have made microvascular anastomosis possible and practical. At present free intestinal interposition grafts and revascularization of the proximal end of pedicle grafts offer the surgeon new options. From 1983 through 1989, reconstruction with vascular anastomosis was performed in 18 cases with pharyngoesophageal carcinoma. Patients were 47 to 88 years old, mean 62.1 years. Reconstructive methods included; free jejunal graft in 14, revascularization of proximal end of a pedicled gastric tube in 1, revascularization of proximal end of a pedicled jejunum in 1, interposition of free jejunal graft between a pedicled gastric tube and the pharynx in 1 and interposition of free ileal graft between a pedicled jejunum and the cervical esophagus in 1 case. Modified radical neck dissection was performed in all cases and superior mediastinal lymph node dissection with median sternotomy was added in 5. The internal jugular vein and common carotid artery were the donor vessels of choice, but in the case of severe sclerosis of the carotid artery, another cervical artery was chosen. The vascular anastomosis was performed using interrupted 7-0 Prolene sutures in the anterior wall and running 7-0 Prolene sutures in the posterior wall under 2.25 magnification. The patency of the anastomosed vessels was confirmed by Doppler flowmeter. The postoperative following-up periods ranged from 1 to 73 months with a mean of 16.5 months, and the survival rate was 66.2% at 1 year and 42.6% after 3 years.(ABSTRACT TRUNCATED AT 250 WORDS)


Flibanserin: a potential treatment for Hypoactive Sexual Desire Disorder in premenopausal women.
Hypoactive Sexual Desire Disorder (HSDD) is defined as a persistent or recurrent deficiency of sexual fantasies and desire for sexual activity, which causes marked personal distress or interpersonal difficulty, and is not better accounted for by another psychiatric disorder or the direct physiological effects of a substance (e.g., a medication) or medical condition. HSDD is believed to be the most common form of Female Sexual Dysfunction and is associated with emotional distress and relationship problems. No pharmacologic therapy is approved for the treatment of HSDD in premenopausal or naturally postmenopausal women. Flibanserin is a 5-HT(1A) agonist/5-HT(2A) antagonist that is under investigation as a treatment for HSDD in women. The aim of this article is to present an overview of the pharmacology, clinical efficacy and safety of flibanserin. Flibanserin is an investigational drug that is not licensed for any indication in any country.


Adherence to prescribed antiretroviral therapy in human immunodeficiency virus-infected children in the PENTA 5 trial.
Most studies of adherence to highly active antiretroviral therapy in children have been retrospective or cross-sectional. Factors relating to the caregiver, the child and the medication are all considered to be important for good adherence. Adherence with taking prescribed medication was assessed by questionnaires completed at 4, 12, 24 and 48 weeks by caregivers of previously untreated HIV-infected children participating in the PENTA 5 trial, which was designed to evaluate different dual nucleoside reverse transcriptase inhibitor therapy combinations with and without the protease inhibitor nelfinavir. The effects of several factors on adherence and the effect of adherence on virologic suppression were assessed by multivariate logistic regression. Caregivers returned 266 questionnaires including at least 1 for 108 (84%) children in the trial. Nelfinavir was reported to be the most difficult drug to take (38% of questionnaires), but the difficulty decreased over time, P = 0.02. Comments on difficulties in taking and remembering drugs related to fear of disclosure and to unpleasant characteristics of the drugs. Full adherence was reported in 74% of questionnaires, did not change over time and was reported more frequently in children older than 10 years and those with symptomatic HIV disease. More children reporting full adherence achieved HIV RNA <400 copies/ml (e.g. at 48 weeks 79% vs. 50% reporting some nonadherence; overall P = 0.01). Good adherence with taking prescribed medication was associated with virologic response. Social factors were important in explaining nonadherence.


Ara h 2 cross-linking catalyzed by MTGase decreases its allergenicity.
Peanuts, whose major allergen is Ara h 2, are included among the eight major food allergens. After reduction using dithiothreitol (DTT), cross-linking of Ara h 2 could be catalyzed by microbial transglutaminase (MTGase), a widely used enzyme in the food industry. In this study, Ara h 2 cross-linking was catalyzed by MTGase after it was reduced by DTT. Using mass spectrometry and PLINK software, five cross-linkers were identified, and five linear allergen epitopes were found to be involved in the reactions. The IgE binding capacity of cross-linked Ara h 2 was found to be significantly lower compared to that of native and reduced Ara h 2. After simulated gastric fluid (SGF) digestion, the digested products of the cross-linked Ara h 2, again, had a significantly lower IgE binding capacity compared to untreated and reduced Ara h 2. Furthermore, reduced and cross-linked Ara h 2 (RC-Ara h 2) induced lower sensitization in mice, indicating its lower allergenicity. Reduction and MTGase-catalyzed cross-linking are effective methods to decrease the allergenicity of Ara h 2. The reactions involved linear allergen epitopes destroying the material basis of the allergenicity, and this might develop a new direction for protein desensitization processes.


Similar task features shape judgment and categorization processes.
The distinction between similarity-based and rule-based strategies has instigated a large body of research in categorization and judgment. Within both domains, the task characteristics guiding strategy shifts are increasingly well documented. Across domains, past research has observed shifts from rule-based strategies in judgment to similarity-based strategies in categorization, but limited these comparisons to 1 prototypical environment, a linear task structure, and a restricted set of strategies. To systematically compare the 2 domains, we considered several instantiations of rule-based and similarity-based strategies and examined strategy choice across different types of judgment and categorization tasks. Between participants, we varied task characteristics from a 1-dimensional linear to a multidimensional linear and to 2 multidimensional nonlinear tasks. Irrespective of domain, strategies considered, or model comparison technique used, we find that more participants relied on similarity-based strategies when the functional relationship between the cues and the criterion was nonlinear. Shifts from rule-based strategies in judgment to similarity-based strategies in categorization, however, were rare and most pronounced in 1-dimensional environments. These results support the hypothesis that the cognitive strategies people select to solve a judgment or categorization task depend less on the domain but more on the complexity of the task. (PsycINFO Database Record


Weak and contradictory effects of self-medication with nectar nicotine by parasitized bumblebees.
The presence of antimicrobial secondary metabolites in nectar suggests that pollinators, which are threatened globally by emergent disease, may benefit from the consumption of nectars rich in these metabolites. We tested whether nicotine, a nectar secondary metabolite common in Solenaceae and Tilia species, is used by parasitized bumblebees as a source of self-medication , using a series of toxicological, microbiological and behavioural experiments. Caged bees infected with Crithidia bombi [TI1] had a slight preference for sucrose solution laced with the alkaloid and behavioural tests showed that the parasite infection induced an increased consumption of nicotine during foraging activity. When ingested, nicotine delayed the progression of a gut infection in bumblebees by a few days, but dietary nicotine did not clear the infection, and after 10 days the parasite load approached that of control bees. Moreover, when pathogens were exposed to the alkaloid prior to host ingestion the protozoan's viability was not directly affected, suggesting that anti-parasite effects were relatively weak. Nicotine consumption in a single dose did not impose any cost even in food-stressed bees (starved) but the alkaloid had detrimental effects on healthy bees if consistently consumed for weeks. These toxic effects disappeared in infected bees suggesting that detoxification costs might have been counterbalanced by the advantages in slowing the progression of the infection. Nonetheless we did not find a benefit of nicotine consumption in terms of life expectancy of infected bees, making these findings difficult to interpret. Our results indicate that caution is warranted in interpreting impacts of plant metabolites on insect parasites and suggest that the conditions under which nicotine consumption provides benefits to either bees or plants remain to be identified. The contention that secondary metabolites in nectar may be under selection from pollinators, or used by plants to enhance their own reproductive success, remains to be confirmed.


Tenofovir Plasma Concentration from Preexposure Prophylaxis at the Time of Potential HIV Exposure: a Population Pharmacokinetic Modeling and Simulation Study Involving Serodiscordant Couples in East Africa.
The Partners Demonstration Project was a prospective, open-label, implementation science-driven study of preexposure prophylaxis (PrEP) among heterosexual HIV serodiscordant couples in Kenya and Uganda. Adherence data were collected using the Medication Event Monitoring System (MEMS), and time of sexual activity was collected using the mobile phone short message service (SMS). Two plasma samples were collected at a single study visit. We integrated adherence, pharmacokinetics, and SMS data using a population pharmacokinetic (PopPK) model to simulate tenofovir plasma concentrations from PrEP at the time of sexual activity. In the first stage of this analysis, we used data from the current study to update a prior PopPK model of tenofovir (TFV) developed with data from the Partners PrEP Study (a phase III clinical trial). The second stage involved simulating plasma concentrations at the time of sexual activity using empirical Bayes estimates (EBEs) derived from the final model. In addition, EBEs from a previously published parent metabolite model of TFV (MTN-001, an open-label 3-way crossover study in healthy women) was used to simulate tenofovir diphosphate (TFV-DP) concentrations. We estimated percent PrEP "coverage" as the number of reported sexual events during which simulated concentrations were above an a priori threshold concentrations associated with a high degree of protection from HIV infection: plasma TFV of >40 ng/ml and peripheral blood mononuclear cell (PBMC) TFV-DP concentration of >36 fmol/million cells. The levels of coverage were 72% for TFV and 81% for TFV-DP. These levels are consistent with a high degree of protection against HIV acquisition in this study of a pragmatic delivery model for antiretroviral-based HIV prevention.


Reliable self-replicating machines in asynchronous cellular automata.
We propose a self-replicating machine that is embedded in a two-dimensional asynchronous cellular automaton with von Neumann neighborhood. The machine dynamically encodes its shape into description signals, and despite the randomness of cell updating, it is able to successfully construct copies of itself according to the description signals. Self-replication on asynchronously updated cellular automata may find application in nanocomputers, where reconfigurability is an essential property, since it allows avoidance of defective parts and simplifies programming of such computers.


The OKRA leaf shape mutation in cotton is active in all cell layers of the leaf.
Okra (L2O) is a semidominant mutation of cotton (Gossypium barbadense) that alters leaf shape by increasing the length of lobes and decreasing lamina expansion. Chimeras containing L2O and wild-type tissue were generated using Semigamy (Se), a mutation that blocks syngamy during fertilization and produces haploid maternal/paternal chimeral progeny at low frequency. In sectorial chimeras, changes in leaf morphology coincide with the boundary between mutant and wild-type tissues, suggesting that L2O does not regulate a laterally diffusible factor within the leaf. However, in mericlinal or periclinal chimeras, the presence of L2O in tissue derived from any of the three histogenic layers (L1, L2, or L3) of the shoot apical meristem produced leaves with a partial mutant phenotype. The presence of L2O in the epidermis (an L1 derivative), or in the subepidermal mesophyll of the leaf (L2 derivatives) reduced the growth of the lamina and thus increased the depth of leaf lobes. The presence of L2O in the middle mesophyll of the lamina and the vasculature of major lateral veins (L3 derivatives) had no local effect on the expansion of the lamina, but significantly increased lobe length. These results demonstrate that L2O is active in every tissue layer of the leaf.


KSR1 is coordinately regulated with Notch signaling and oxidative phosphorylation in thyroid cancer.
Kinase suppressor of RAS1 (KSR1) is a scaffold protein implicated in RAS-mediated RAF activation. However, the molecular function of KSR in papillary thyroid cancer (PTC) is unknown. Thus, this study aimed to characterize the role of KSR1 in patients with PTC. qRT-PCR and immunohistochemistry (IHC) revealed inter-tumor heterogeneities in the expression of KSR1 in PTC tissues. Interestingly, BRAFV600E-positive PTC showed higher KSR1 mRNA expression than BRAFV600E-negative PTC (P<0.001). Gene Set Enrichment Analysis (GSEA) using public repositories showed that high KSR1 expression coordinately upregulated Notch signaling (nominal P=0.019, false discovery rate (FDR) q-value=0.165); this finding was supported by GeneNetwork analysis, indicating that KSR1 expression is positively correlated with NOTCH1 expression (ρ=0.677, P=6.15×10(-9)). siRNA against KSR1 (siKSR1) significantly decreased ERK phosphorylation induced by BRAFV600E, resulting in reduced expression of NOTCH1 and HES1, targets of Notch signaling. GSEA revealed that high KSR1 expression was also associated with downregulation of genes related to oxidative phosphorylation (OxPhos). Consistent with this, electron microscopy showed that PTCs with high KSR1 expression exhibited structural defects of the mitochondrial cristae. Furthermore, siKSR1-transfected BCPAP and 8505C cells generated fewer colonies in colony-forming assays. In addition, GSEA showed that high expression of KSR2 and connector enhancer of KSR1 (CNKSR1) also coordinately upregulated Notch signaling (KSR2: nominal P=0.0097, FDR q-value=0.154 and CNKSR1: nominal P<0.0001, FDR q-value=0.00554), and high CNKSR2 was associated with downregulation of the OxPhos gene set (nominal P<0.0001, FDR q-value <0.0001). In conclusion, KSR1 is coordinately regulated with Notch signaling and OxPhos in PTC, because its scaffold function might be required to sustain the proliferative signaling and metabolic remodeling associated with this type of cancer.


Can Human Embryonic Stem Cell-Derived Stromal Cells Serve a Starting Material for Myoblasts?
A large number of myocytes are necessary to treat intractable muscular disorders such as Duchenne muscular dystrophy with cell-based therapies. However, starting materials for cellular therapy products such as myoblasts, marrow stromal cells, menstrual blood-derived cells, and placenta-derived cells have a limited lifespan and cease to proliferate in vitro. From the viewpoints of manufacturing and quality control, cells with a long lifespan are more suitable as a starting material. In this study, we generated stromal cells for future myoblast therapy from a working cell bank of human embryonic stem cells (ESCs). The ESC-derived CD105+ cells with extensive in vitro proliferation capability exhibited myogenesis and genetic stability in vitro. These results imply that ESC-derived CD105+ cells are another cell source for myoblasts in cell-based therapy for patients with genetic muscular disorders. Since ESCs are immortal, mesenchymal stromal cells generated from ESCs can be manufactured at a large scale in one lot for pharmaceutical purposes.


Fine-scale mapping using Hardy-Weinberg disequilibrium.
Hardy-Weinberg disequilibrium (HWD) among affected individuals has recently been proposed for fine-scale mapping of disease susceptibility genes. We investigate the statistical properties of several available HWD measures and develop a new HWD measure J for fine-scale mapping. It is shown both theoretically and through simulations that the available HWD measures depend not only on the genetic distance between the marker locus of interest and the disease susceptibility locus, but also on the allele frequencies at the marker locus. On the contrary, the new measure is not affected by the allele frequencies at the marker locus under the following assumptions: (a) there is initial complete linkage disequilibrium between the marker and the disease loci, (b) there are no new mutations at the marker and the disease loci, and (c) the population under study is large. We develop a novel method to estimate the location of the disease susceptibility gene based on the HWD measure J. The estimator is robust to low mutation rates at the marker and the disease loci. We compare the standard error of the estimated disease gene loci using P excess for case-control studies with the standard error using J for case-only studies under various disease models. The newly developed method is successfully applied to a data set on hereditary haemochromatosis (HH).


Effects of climate warming on carbon fluxes in grasslands- A global meta-analysis.
Climate warming will affect terrestrial ecosystems in many ways, and warming-induced changes in terrestrial carbon (C) cycling could accelerate or slow future warming. So far, warming experiments have shown a wide range of C flux responses, across and within biome types. However, past meta-analyses of C flux responses have lacked sufficient sample size to discern relative responses for a given biome type. For instance grasslands contribute greatly to global terrestrial C fluxes, and to date grassland warming experiments provide the opportunity to evaluate concurrent responses of both plant and soil C fluxes. Here, we compiled data from 70 sites (in total 622 observations) to evaluate the response of C fluxes to experimental warming across three grassland types (cold, temperate, and semi-arid), warming methods, and short (≤3 years) and longer-term (>3 years) experiment lengths. Overall, our meta-analysis revealed that experimental warming stimulated C fluxes in grassland ecosystems with regard to both plant production (e.g., net primary productivity (NPP) 15.4%; aboveground NPP (ANPP) by 7.6%, belowground NPP (BNPP) by 11.6%) and soil respiration (Rs) (9.5%). However, the magnitude of C flux stimulation varied significantly across cold, temperate and semi-arid grasslands, in that responses for most C fluxes were larger in cold than temperate or semi-arid ecosystems. In semi-arid and temperate grasslands, ecosystem respiration (Reco) was more sensitive to warming than gross primary productivity (GPP), while the opposite was observed for cold grasslands, where warming produced a net increase in whole-ecosystem C storage. However, the stimulatory effect of warming on ANPP and Rs observed in short-term studies (≤3 years) in both cold and temperate grasslands disappeared in longer-term experiments (>3 years). These results highlight the importance of conducting long-term warming experiments, and in examining responses across a wide range of climate.


Peri-operative efficacy and long-term survival benefit of robotic-assisted radical cystectomy in septuagenarian patients compared with younger patients: a nationwide multi-institutional study in Japan.
To determine the peri-operative safety and oncological value of robotic-assisted radical cystectomy (RARC) for older and younger patients in an initial Japanese RARC series. We retrospectively analyzed the demographics, complications, peri-operative and oncological outcomes of 253 consecutive patients with bladder cancer who underwent RARC at 34 institutions in Japan between April 2009 and March 2017. The patients were assigned to groups according to ages at surgery of < 70 (younger; n = 125) and ≥ 70 (older; n = 128) years. Mean Charlson comorbidity index (p = 0.045) and the incidence of a history of previous abdominal surgery (p = 0.002) were significantly higher, whereas a history of neoadjuvant chemotherapy (p = 0.028) and neobladder (p < 0.001) were significantly lower in the older group. Mean total operative time was significantly shorter (p = 0.019) and mean estimated blood loss (p = 0.013) was significantly lower in the older group. Post-operative Grade ≥ II complications were comparable at 0-30, 31-90 and 91 days after surgery despite urinary tract associations. Rates of positive surgical margins and mean numbers of removed lymph nodes were comparable between the two groups. Although 5-year overall survival rates were significantly lower (p = 0.03) for older patients, 5-year cancer-specific (p = 0.10) and recurrence-free survival rates were comparable (p = 0.20) between the groups. Using RARC potentially allows the application of less invasive procedures and cancer control for septuagenarian patients that are equivalent to those for younger patients.