Changes in body habitus and serum lipid abnormalities in HIV-positive women on highly active antiretroviral therapy (HAART).
Twenty-one women (propositi) who expressed serious concerns about changes in body habitus during highly active antiretroviral therapy (HAART) were evaluated by thorough physical examination, anthropometric measurements, and serum lipid and endocrine assays. The same evaluations were carried out in a comparison group of 21 women who received HAART but did not complain of changes in habitus. No significant demographic differences were found between the propositi and the comparison group, nor were there significant differences in CD4 count or plasma viral load (PVL) between the two groups. Lipid analyses were also performed on plasma obtained prior to HAART from 12 of the women. The frequency of changes reported by the 21 propositi were increase in abdominal size (90%), increase in breast size (71%), weight gain of >5 kg (43%), peripheral fat wasting (43%), buttock fat wasting (38%) and development of cervicodorsal fat pad (19%). A subset of patients in the comparison group experienced increase in abdominal size (29%) and weight gain >5 kg (19%), but none experienced clinically detectable peripheral or buttock fat wasting, increased breast size, or development of cervicodorsal fat pads. Mean waist circumference, waist-to-hip ratios (WHR), body fat, and body mass index (BMI) were above the desirable range for women in both propositi and the comparison group. Levels of total cholesterol, triglycerides, low-density lipoprotein (LDL) cholesterol, and high-density lipoprotein (HDL) cholesterol associated with increased cardiovascular risk were found in 48%, 62%, 45%, and 33%. respectively, of the propositi, with similar findings in the comparison group. Fasting insulin levels were elevated in 4 propositi and 6 of the comparison group; mean insulin levels were within the normal range for both groups. In the comparison of lipids for the subset of patients before and after HAART therapy, HAART was associated with significant increases in total cholesterol, apolipoprotein B, and HDL cholesterol. Changes in body habitus caused by redistribution of fat occur commonly in women receiving HAART. Serum lipid abnormalities also are common during HAART and appear to be as frequent in women who do not experience clinically apparent body fat redistribution as in those who do. The observed changes in body fat distribution and in serum lipid levels are alterations that have been strongly correlated with increased risk for cardiovascular disease. Therefore, an understanding of the basis of these phenomena, and the risks with which they may be associated in this population, will be important for therapeutic decision making in women with HIV disease.


Arterial false aneurysms and their modern management.
Both diagnostic and therapeutic options in the management of iatrogenic false aneurysms have changed dramatically in the last decade, with surgery being required only rarely. To describe our experience, techniques and results in treating pseudoaneurysms at a large medical center with frequent arterial interventions. We emphasize upper limb lesions. We reviewed the data of all consecutive patients diagnosed by color-coded duplex Doppler between August 1992 and July 1998 as having upper limb and lower limb pseudoaneurysms (mainly post-catheterization). We accumulated 107 false aneurysms (mainly post-catheterization lesions): 5 were upper limb lesions and 102 were groin aneurysms. In the lower limb cases 94 of the 102 lesions were not operated upon (92.1%). Seventy lower limb cases were treated non-operatively by ultrasound-guided compression obliteration with a 95.7% success rate (67 cases). Two cases were treated by percutaneous thrombin injection (2%) and 23 by observation only (22.5%). Altogether 12 patients underwent surgery (11.2%): 4 upper extremity and 8 lower extremity cases. None of the lower limb group suffered serious complications regardless of treatment, but all five upper limb cases did, four of them necessitating surgical intervention. Three of the five upper limb cases had a grave outcome with severe or permanent functional or neurological damage. Most post-catheterization pseudoaneurysms can be managed non-surgically. False aneurysms in the upper extremity are rare, comprising less than 2% of all lesions. However, upper extremity pseudoaneurysms present a potentially more serious complication and require early diagnosis and prompt intervention to minimize the high complication rate and serious long-term sequelae. Prevention can be achieved by proper puncture technique and site selection, and correct post-procedure hemostatic compression with or without an external device. Some upper limb lesions are avoidable if the axillary artery is not punctured.


Use of guideline-recommended therapies for heart failure in the Medicare population.
Most information about the use of guideline-recommended therapies for heart failure reflects what occurred at discharge after an inpatient stay. Using a nationally representative, community-dwelling sample of elderly Medicare beneficiaries, we examined how the use of angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockers (ARBs), and beta-blockers has changed and factors associated with their use. Using data from the Medicare Current Beneficiary Survey cost and use files matched with Medicare claims data, we identified beneficiaries for whom a diagnosis of heart failure was reported between January 1, 2000, and December 31, 2004. Data on medications prescribed during the year of cohort entry were based on patient self-report. We used multivariable logistic regression to explore relationships between the use of ACE inhibitors/ARBs and beta-blockers and patient demographic characteristics. From 2000 through 2004, the use of ARBs increased from 12% to 19%, and the use of beta-blockers increased from 30% to 41%. The use of ACE inhibitors remained constant at 45%. Beneficiaries who reported having prescription drug insurance coverage were 32% more likely than other beneficiaries to have filled a prescription for an ACE inhibitor or ARB and 26% more likely to have filled a prescription for a beta-blocker. Although the use of guideline-recommended therapies for heart failure has increased, it remains suboptimal.


Retrospective Analysis of Esophageal Heat Transfer for Active Temperature Management in Post-cardiac Arrest, Refractory Fever, and Burn Patients.
Core temperature management is an important aspect of critical care; preventing unintentional hypothermia, reducing fever, and inducing therapeutic hypothermia when appropriate are each tied to positive health outcomes. The purpose of this study is to evaluate the performance of a new temperature management device that uses the esophageal environment to conduct heat transfer. De-identified patient data were aggregated from three clinical sites where an esophageal heat transfer device (EHTD) was used to provide temperature management. The device was evaluated against temperature management guidelines and best practice recommendations, including performance during induction, maintenance, and cessation of therapy. Across all active cooling protocols, the average time-to-target was 2.37 h and the average maintenance phase was 22.4 h. Patients spent 94.9% of the maintenance phase within ±1.0°C and 67.2% within ±0.5°C (574 and 407 measurements, respectively, out of 605 total). For warming protocols, all of the patient temperature readings remained above 36°C throughout the surgical procedure (average 4.66 h). The esophageal heat transfer device met performance expectations across a range of temperature management applications in intensive care and burn units. Patients met and maintained temperature goals without any reported adverse events.


Indicators for assessing folate and vitamin B-12 status and for monitoring the efficacy of intervention strategies.
Deficiencies of folate or of vitamin B-12 are widespread and constitute a major global burden of morbidity that affect all age groups. Detecting or confirming the presence of folate or vitamin B-12 deficiency and distinguishing one from the other depends, ultimately, on laboratory testing. Tests to determine the presence of folate or vitamin B-12 deficiency are used singly or in combination to establish the nutritional status and prevalence of deficiencies of the vitamins in various populations. The efficacy of interventions through the use of fortification or supplements is monitored by using the same laboratory tests. Tests currently in use have limitations that can be either technical or have a biological basis. Consequently, each single test cannot attain perfect sensitivity, specificity, or predictive value. Laboratory indicators of vitamin B-12 or folate status involve the measurement of either the total or a physiologically relevant fraction of the vitamin in a compartment such as blood. Thus, assays to measure vitamin B-12 or folate in plasma or serum as well as folate in red blood cells are in widespread use, and more recently, methods to measure vitamin B-12 associated with the plasma binding protein transcobalamin (holotranscobalamin) have been developed. Alternatively, concentrations of surrogate biochemical markers that reflect the metabolic function of the vitamin can be used. Surrogates most commonly used are plasma homocysteine, for detection of either vitamin B-12 or folate deficiency, and methylmalonic acid for detection of vitamin B-12 deficiency. The general methods as well as their uses, indications, and limitations are presented.


Controlled electroplating and electromigration in nickel electrodes for nanogap formation.
We report the fabrication of nickel nanospaced electrodes by electroplating and electromigration for nanoelectronic devices. Using a conventional electrochemical cell, nanogaps can be obtained by controlling the plating time alone and after a careful optimization of electrodeposition parameters such as electrolyte bath, applied potential, cleaning, etc. During the process, the gap width decreases exponentially with time until the electrode gaps are completely bridged. Once the bridge is formed, the ex situ electromigration technique can reopen the nanogap. When the gap is ∼ 1 nm, tunneling current-voltage characterization shows asymmetry which can be corrected by an external magnetic field. This suggests that charge transfer in the nickel electrodes depends on the orientation of magnetic moments.


Identifying Social Skills That Support Housing Attainment and Retention Among Homeless Persons With Serious Mental Illness.
By combining supportive services with independent housing, permanent supportive housing (PSH) programs effectively address homelessness. Yet, many persons with serious mental illness struggle to attain and retain housing in these programs. Social skills-which facilitate social interactions and instrumental tasks-predict premature exit from PSH. This project aimed to build consensus on a set of social skills that supports independent housing attainment and retention among homeless persons with serious mental illness, with a clinical focus on PSH programs. Guided by the RAND/University of California, Los Angeles, appropriateness method, the set of social skills was developed by using literature review; key informant interviews (N=12), a national consensus panel of experts in psychosocial rehabilitation and homelessness (N=11), and two focus groups with homeless persons with serious mental illness (N=17). These methods identified 24 social skills in seven domains: finding and renting an apartment, using one's time well, getting closer to people, managing finances, avoiding problems with drugs and alcohol, solving interpersonal problems, and managing one's health. Expert panelists and focus group participants agreed that these social skills were feasible for inclusion in social skills training and could strongly affect housing outcomes in PSH settings. Consensus was reached about a set of social skills relevant to housing attainment and retention for persons with serious mental illness engaged in PSH. Next steps include modifying social skills training interventions to this skill set, making contextual modifications relevant to the setting and context of PSH, and studying the effectiveness and implementation of the adapted intervention in PSH.


Identification of a common lupus disease-associated microRNA expression pattern in three different murine models of lupus.
Recent reports have shown that microRNAs (miRNAs) regulate vital immunological processes and have emerged as key regulators of immune system development and function. Therefore, it is important to determine miRNA dysregulation and its pathogenic contribution in autoimmune diseases, an aspect not adequately addressed thus far. In this study, we profiled miRNA expressions in splenic lymphocytes from three murine lupus models (MRL-lpr, B6-lpr and NZB/W(F₁)) with different genetic background by miRNA microarray assays and Real-time RT-PCR. Despite the genetic differences among these three lupus stains, a common set of dysregulated miRNAs (miR-182-96-183 cluster, miR-31, and miR-155) was identified in splenocytes when compared with age-matched control mice. The association of these miRNAs with the disease was highlighted by our observation that this miRNA expression pattern was evident in NZB/W mice only at an age when lupus disease is manifested. Further, we have shown that the miRNA dysregulation in MRL-lpr mice was not simply due to the activation of splenocytes. By Real-time RT-PCR, we confirmed that these miRNAs were upregulated in both purified splenic B and T cells from MRL-lpr mice. miR-127 and miR-379, which were greatly upregulated in splenocytes from lpr mice, were moderately increased in diseased NZB/W mice. In addition, Real-time RT-PCR revealed that miR-146a, miR-101a, and miR-17-92 were also markedly upregulated in splenic T, but not B cells from MRL-lpr mice. The identification of common lupus disease-associated miRNAs now forms the basis for the further investigation of the pathogenic contribution of these miRNAs in autoimmune lupus, which will advance our knowledge of the role of miRNAs in autoimmunity. Given that miRNAs are conserved, with regard to both evolution and function, our observation of a common lupus disease-associated miRNA expression pattern in murine lupus models is likely to have significant pathogenic, diagnostic, and/or therapeutic implications in human lupus.


Addicted to love: What is love addiction and when should it be treated?
Recent research suggests that romantic love can be literally addictive. Although the exact nature of the relationship between love and addiction has been described in inconsistent terms throughout the literature, we offer a framework that distinguishes between a narrow view and a broad view of love addiction. The narrow view counts only the most extreme, harmful forms of love or love-related behaviors as being potentially addictive in nature. The broad view, by contrast, counts even basic social attachment as being on a spectrum of addictive motivations, underwritten by similar neurochemical processes as more conventional addictions. We argue that on either understanding of love-as-addiction, treatment decisions should hinge on considerations of harm and well-being rather than on definitions of disease. Implications for the ethical use of anti-love biotechnology are considered.


Arthroscopic versus open ankle arthrodesis: a multicenter comparative case series.
Ankle arthrodesis results in measurable improvements in terms of pain and function in patients with end-stage ankle arthritis. Arthroscopic ankle arthrodesis has gained increasing popularity, with reports of shorter hospital stays, shorter time to solid fusion, and equivalent union rates when compared with open arthrodesis. However, there remains a lack of high-quality prospective data. We evaluated the results of open and arthroscopic ankle arthrodesis in a comparative case series of patients who were managed at two institutions and followed for two years. The primary outcome was the Ankle Osteoarthritis Scale score, and secondary outcomes included the Short Form-36 physical and mental component scores, the length of hospital stay, and radiographic alignment. There were thirty patients in each group. Both groups showed significant improvement in the Ankle Osteoarthritis Scale score and the Short Form-36 physical component score at one and two years. There was significantly greater improvement in the Ankle Osteoarthritis Scale score at one year and two years and shorter hospital stay in the arthroscopic arthrodesis group. Complications, surgical time, and radiographic alignment were similar between the two groups. Open and arthroscopic ankle arthrodesis were associated with significant improvement in terms of pain and function as measured with the Ankle Osteoarthritis Scale score. Arthroscopic arthrodesis resulted in a shorter hospital stay and showed better outcomes at one and two years.


Scattering of light by polyhedral ice crystals.
The single-scattering phase functions of polyhedral-shaped ice particles are calculated by means of geometric optics and the diffraction theory. Particle orientation is assumed to be random in space. Particle shapes are taken both from ice-crystal classifications and from in situ measurements. The effects of particle concavity on the scattering signature are discussed in detail. A common feature is the pronounced forward-scattering peak, as well as different halo peaks that are due to a minimum deviation at corresponding ice prisms. An unusual halo phenomena, which results from a minimum deviation in a double-prism configuration, is found and verified. The comparison of different particle types shows that backscattering is a sensitive indicator for the identification of types of ice-crystal. Aggregate particles, like bullet rosettes, basically show the scattering characteristics of their individual components.


Diffuse spreading Clostridium septicum infection, malignant disease and immune suppression.
Nineteen patients have been treated with gas gangrene infection proved on cultures to be caused by Clostridium septicum. Infection occurred after trauma in three patients and occurred spontaneously in 16, including six patients with peritonitis. None of the instances occurred as a postoperative infection. Ten of the patients had an associated malignant disease including carcinoma of the colon and rectum in six, hematologic malignant conditions in three and carcinoma of the gallbladder in one. Of the remaining six patients, four had evidence of immune suppression (two had leukopenia, one was preleukemic and one had undergone a postcadaver renal transplant), one had diabetes and one had overwhelming sepsis due to a perforated gallbladder with peritonitis. Only three of the 19 patients were otherwise healthy, and infection occurred after trauma in all of these patients. Four patients were moribund upon admission and died before initial therapy was completed. In the other 15 patients, treatment consisted of administration of penicillins and broad spectrum antibiotics and débridement, as well as hyperbaric oxygen in 13 patients. Thirteen of 19 patients died and eight of ten with malignant disease died. The data show a striking relationship between Clostridium septicum and both malignant disease and immune suppression.


Dental caries prevalence among type II diabetic and nondiabetic adults attending a hospital.
Diabetes mellitus (DM) is a common chronic metabolic disorder which affects millions of people. At present, India has the highest incidence of diabetes worldwide. Several oral lesions and conditions are associated with diabetes. However, there is a lack of consensus among researchers regarding the relationship between DM and dental caries. Hence, the present study was carried out to assess the dental caries prevalence among type II diabetic and nondiabetic adults attending a hospital in Ahmedabad city. A hospital-based cross-sectional study was conducted. One hundred and twenty diabetics individuals attending the diabetic Outpatient Department (OPD) and age and sex-matched 120 nondiabetic individuals from general OPD were included in the study. The data were gathered through semi-close-ended questionnaire and clinical examination. Dental caries was assessed by using the World Health Organization's 2013 proforma. Data was analyzed by applying Student's independent t-test or one-way analysis of variance. Dental caries prevalence among the diabetic group was 73.33% and 33.33% among the nondiabetic group. Dental caries prevalence and mean dental caries was significantly higher among uncontrolled diabetic individuals than that among controlled diabetic individuals. Duration of the disease and dental caries prevalence did not show any significant difference. Dental caries prevalence was significantly high among diabetic individuals compared with nondiabetic individuals. Close collaboration between the patients, healthcare units, and oral health professionals could be a way of improving diabetic patients' general and oral health.


Non-standard vision measures predict mortality in elders: the Smith-Kettlewell Institute (SKI) study.
To determine which vision tests predict mortality within 10 years in a community-based elderly sample. Nine hundred residents of Marin County, California 58 to 101 years of age (mean 75 years at baseline), underwent a battery of tests, including high contrast acuity, low contrast acuity, low contrast/low luminance acuity, acuity in glare, contrast sensitivity, color vision, stereopsis, standard and attentional fields. The association between the vision tests and mortality within 10 years of baseline was assessed with Cox Proportional Hazards models controlling for age, sex, education level, depression, cognitive status and self-reported medical conditions. Forty-three percent of the sample died within 10 years of baseline. When controlling for mortality-related covariates, impairment in any of the vision measures was associated with increased risk of death. However, non-standard vision measures (ie, impairment in low contrast/low luminance acuity, standard field integrity and the impact of the attentional task on field integrity) were more highly associated with mortality than standard high contrast acuity. In agreement with other studies, we find that visual impairment is a significant predictor of death. However, the strongest relationship was found for measures other than high contrast acuity. These results suggest that non-standard vision measures may be more sensitive indicators of generalized aging in the most elderly.


Client perceptions of the quality of primary care services in Afghanistan.
To identify factors associated with client perceptions of the quality of primary care services in Afghanistan. Cross-sectional survey of outpatient health facilities, health workers, patients and caretakers. Primary health care facilities in every province of Afghanistan. Main outcome measure Numerical scale of client perceptions of service quality. Clients report relatively high levels of perceived quality in Afghanistan. Most of the variation that is explained relates specifically to the patient's interaction with the health worker and not to other health facility characteristics, such as cleanliness, infrastructure, service capacity and the presence of equipment or drugs. The strongest determinants of client-perceived quality identified are health worker thoroughness in taking patient histories, conducting physical examinations and communicating with patients. Being seen by a doctor and being from a household in the poorest quintile are also associated with higher perceived quality. For female patients, being seen by a female provider is associated with higher perceived quality, while for male patients time and money spent for travel to the health facility are negatively associated with perceived quality. Clinical quality and client perceived quality appear to be mutually reinforcing, and efforts to improve health worker performance in taking histories, conducting exams and communicating with patients are likely to increase client perceived quality in this setting. Client perceptions of service quality assume additional importance in Afghanistan, where the perceived legitimacy of the government may depend partially on its ability to convince the population that it can deliver essential health services.


Nuclear receptors in disease: the oestrogen receptors.
For several decades, it has been known that oestrogens are essential for human health. The discovery that there are two oestrogen receptors (ERs), ERalpha and ERbeta, has facilitated our understanding of how the hormone exerts its physiological effects. The ERs belong to the family of ligand-activated nuclear receptors, which act by modulating the expression of target genes. Studies of ER-knockout (ERKO) mice have been instrumental in defining the relevance of a given receptor subtype in a certain tissue. Phenotypes displayed by ERKO mice suggest diseases in which dysfunctional ERs might be involved in aetiology and pathology. Association between single-nucleotide polymorphisms (SNPs) in ER genes and disease have been demonstrated in several cases. Selective ER modulators (SERMs), which are selective with regard to their effects in a certain cell type, already exist. Since oestrogen has effects in many tissues, the goal with a SERM is to provide beneficial effects in one target tissue while avoiding side effects in others. Refined SERMs will, in the future, provide improved therapeutic strategies for existing and novel indications.


Oligopeptide permease in Borrelia burgdorferi: putative peptide-binding components encoded by both chromosomal and plasmid loci.
To elucidate the importance of oligopeptide permease for Borrelia burgdorferi, the agent of Lyme disease, a chromosomal locus in B. burgdorferi that encodes homologues of all five subunits of oligopeptide permease has been identified and characterized. B. burgdorferi has multiple copies of the gene encoding the peptide-binding component, OppA; three reside at the chromosomal locus and two are on plasmids. Northern analyses indicate that each oppA gene is independently transcribed, although the three chromosomal oppA genes are also expressed as bi- and tri-cistronic messages. Induction of one of the plasmid-encoded oppA genes was observed following an increase in temperature, which appears to be an important cue for adaptive responses in vivo. The deduced amino acid sequences suggest that all five borrelial oppA homologues are lipoproteins, but the protease-resistance of at least one of them in intact bacteria is inconsistent with outer-surface localization. Insertional inactivation of a plasmid-encoded oppA gene demonstrates that it is not essential for growth in culture.


Mapping malaria transmission intensity in Malawi, 2000-2010.
Substantial development assistance has been directed towards reducing the high malaria burden in Malawi over the past decade. We assessed changes in transmission over this period of malaria control scale-up by compiling community Plasmodium falciparum rate (PfPR) data during 2000-2011 and used model-based geostatistical methods to predict mean PfPR2-10 in 2000, 2005, and 2010. In addition, we calculated population-adjusted prevalences and populations at risk by district to inform malaria control program priority setting. The national population-adjusted PfPR2-10 was 37% in 2010, and we found no evidence of change over this period of scale-up. The entire population of Malawi is under meso-endemic transmission risk, with those in districts along the shore of Lake Malawi and Shire River Valley under highest risk. The lack of change in prevalence confirms modeling predictions that when compared with lower transmission, prevalence reductions in high transmission settings require greater investment and longer time scales.


Functional interdependence of DNA polymerizing and 3'-->5' exonucleolytic activities in Pyrococcus furiosus DNA polymerase I.
Pyrococcus furiosus DNA polymerase I (Pol BI) belongs to the family B (alpha-like) DNA polymerases and has a strong 3'-->5' exonucleolytic activity, in addition to its DNA polymerizing activity. To understand the relationship between the structure and function of this DNA polymerase, three deletion mutants, Delta1 (DeltaLeu746-Ser775), Delta2 (DeltaLeu717-Ser775) and Delta3 (DeltaHis672-Ser775), and two substituted mutants of Asp405, D405A and D405E, were constructed. These substitutions affected both the DNA polymerizing and the 3'-->5' exonucleolytic activities. The Delta1 mutant protein had DNA polymerizing activity with higher specific activity than that of the wild-type Pol BI, but retained only 10% of the exonucleolytic activity of the wild-type. The other two deletion mutants lost most of both activities. These results suggest that the DNA polymerizing and exonucleolytic activities are closely related to each other in the folded structure of this DNA polymerase, as proposed in the family B DNA polymerases.


Genome-based proteomics.
Protein-protein interactions play crucial roles in various biological pathways and functions. Therefore, the characterization of protein levels and also the network of interactions within an organism would contribute considerably to the understanding of life. The availability of the human genome sequence has created a range of new possibilities for biomedical research. A crucial challenge is to utilize the genetic information for better understanding of protein distribution and function in normal as well as in pathological biological processes. In this review, we have focused on different platforms used for systematic genome-based proteome analyses. These technologies are in many ways complementary and should be seen as various ways to elucidate different functions of the proteome.


Aortic root dilatation in adults with surgically repaired tetralogy of fallot: a multicenter cross-sectional study.
Although aortic root pathology has been described in patients with tetralogy of Fallot, the scope of the problem remains poorly defined. We sought to determine the prevalence and predictors of aortic root dilatation in adults with repaired tetralogy of Fallot. A multicenter cross-sectional study was conducted with standardized reassessment of echocardiographic parameters in 474 adults (≥18 years) with surgically repaired tetralogy of Fallot or pulmonary atresia with ventricular septal defect. The aortic root was measured in a parasternal long-axis view, in diastole, at the level of the sinus of Valsalva. Prevalence and predictors of an absolute diameter ≥40 mm and of an observed-to-expected ratio >1.5 were assessed. The aortic root dimension was ≥40 mm in 28.9% (95% confidence interval, 26.9%-30.9%). In multivariate analyses, the only independently associated factor was male sex (odds ratio, 4.48; 95% confidence interval, 1.55-12.89; P=0.006). The prevalence of an observed-to-expected aortic root dimension ratio >1.5 was 6.6% (95% confidence interval, 5.3%-7.9%). It was associated with pulmonary atresia and moderate or severe aortic regurgitation in univariate analyses, but no independent predictor was identified. The side of the aortic arch was not associated with aortic root dimension. The prevalence of moderate or severe aortic regurgitation was 3.5% (95% confidence interval, 2.7%-4.2%). Although nearly one third of adults with repaired tetralogy of Fallot have an aortic root diameter ≥40 mm, the prevalence of a dilated aortic root, when defined by an indexed ratio of observed-to-expected values, is low. Similarly, moderate or severe aortic regurgitation is uncommon.


Buccal absorption of diazepam is improved when administered in bioadhesive tablets-An in vivo study in conscious Göttingen mini-pigs.
Buccal delivery may be clinically beneficial for compounds with a high gastrointestinal and hepatic first pass metabolism or in situations where a fast systemic absorption is desired. The delivery of a crystalline low soluble compounds, e.g. diazepam, may be limited due to the low volume of saliva available to facilitate solvation in order to drive the permeation of drug through the buccal mucosa. Therefore, the present study investigated the potential benefits of administering diazepam either as an amorphous or as a crystalline form in mucoadhesive tablets to conscious Göttingen mini-pigs. Presentation of the compound in the amorphous form lead to a very fast absorption, however, the obtained bioavailability was at the same level observed following buccal administration of a commercially immediate release tablet. Addition of chitosan, as a mucoadhesive excipient, resulted in a higher absolute bioavailability compared to tablets without chitosan. The absorption rate for the chitosan-based tablets was significant slower, probably due to the slower diffusion of the compound out of the tablet. In vitro release data was able to predict the variations in tmax, but otherwise no correlation could be found between in vitro and in vivo data.


Remote sensing of aquatic vegetation distribution in Taihu Lake using an improved classification tree with modified thresholds.
Classification trees (CT) have been used successfully in the past to classify aquatic vegetation from spectral indices (SI) obtained from remotely-sensed images. However, applying CT models developed for certain image dates to other time periods within the same year or among different years can reduce the classification accuracy. In this study, we developed CT models with modified thresholds using extreme SI values (CT(m)) to improve the stability of the models when applying them to different time periods. A total of 903 ground-truth samples were obtained in September of 2009 and 2010 and classified as emergent, floating-leaf, or submerged vegetation or other cover types. Classification trees were developed for 2009 (Model-09) and 2010 (Model-10) using field samples and a combination of two images from winter and summer. Overall accuracies of these models were 92.8% and 94.9%, respectively, which confirmed the ability of CT analysis to map aquatic vegetation in Taihu Lake. However, Model-10 had only 58.9-71.6% classification accuracy and 31.1-58.3% agreement (i.e., pixels classified the same in the two maps) for aquatic vegetation when it was applied to image pairs from both a different time period in 2010 and a similar time period in 2009. We developed a method to estimate the effects of extrinsic (EF) and intrinsic (IF) factors on model uncertainty using Modis images. Results indicated that 71.1% of the instability in classification between time periods was due to EF, which might include changes in atmospheric conditions, sun-view angle and water quality. The remainder was due to IF, such as phenological and growth status differences between time periods. The modified version of Model-10 (i.e. CT(m)) performed better than traditional CT with different image dates. When applied to 2009 images, the CT(m) version of Model-10 had very similar thresholds and performance as Model-09, with overall accuracies of 92.8% and 90.5% for Model-09 and the CT(m) version of Model-10, respectively. CT(m) decreased the variability related to EF and IF and thereby improved the applicability of the models to different time periods. In both practice and theory, our results suggested that CT(m) was more stable than traditional CT models and could be used to map aquatic vegetation in time periods other than the one for which the model was developed.


An in vivo immunotherapy screen of costimulatory molecules identifies Fc-OX40L as a potent reagent for the treatment of established murine gliomas.
We tested the combination of a tumor lysate vaccine with a panel of costimulatory molecules to identify an immunotherapeutic approach capable of curing established murine gliomas. Glioma-bearing mice were primed with a tumor lysate vaccine, followed by systemic administration of the following costimulatory ligands: OX40L, CD80, 4-1BBL, and GITRL, which were fused to the Fc portion of human immunoglobulin. Lymphocytes and mRNA were purified from the brain tumor site for immune monitoring studies. Numerous variations of the vaccine and Fc-OX40L regimen were tested alone or in combination with temozolomide. Lysate vaccinations combined with Fc-OX40L led to the best overall survival, yielding cure rates of 50% to 100% depending on the timing, regimen, and combination with temozolomide. Cured mice that were rechallenged with glioma cells rejected the challenge, showing immunologic memory. Lymphocytes isolated from the draining lymph nodes of vaccine/Fc-OX40L-treated mice had superior tumoricidal function relative to all other groups. Vaccine/Fc-OX40L-treated mice exhibited a significant increase in proliferation of brain-infiltrating CD4 and CD8 T cells, as indicated by Ki67 staining. Fc-OX40L had single-agent activity in transplanted and spontaneous glioma models, and the pattern of inflammatory gene expression in the tumor predicted the degree of therapeutic response. These data show that Fc-OX40L has unique and potent activity against experimental gliomas and warrants further testing.


Connective tissue growth factor-specific monoclonal antibody inhibits growth of malignant mesothelioma in an orthotopic mouse model.
Malignant mesothelioma is an aggressive neoplasm with no particularly effective treatments. We previously reported that overexpression of connective tissue growth factor (CTGF/CCN2) promotes mesothelioma growth, thus suggesting it as a novel molecular target. A human monoclonal antibody that antagonizes CTGF (FG-3019, pamrevlumab) attenuates malignant properties of different kinds of human cancers and is currently under clinical trial for the treatment of pancreatic cancer. This study reports the effects of FG-3019 on human mesothelioma in vitro and in vivo. We analyzed the effects of FG-3019 on the proliferation, apoptosis, migration/invasion, adhesion and anchorage-independent growth in three human mesothelioma cell lines, among which ACC-MESO-4 was most efficiently blocked with FG-3019 and was chosen for in vivo experiments. We also evaluated the coexistent effects of fibroblasts on mesothelioma in vitro, which are also known to produce CTGF in various pathologic situations. Coexistent fibroblasts in transwell systems remarkably promoted the proliferation and migration/invasion of mesothelioma cells. In orthotopic nude mice model, FG-3019 significantly inhibited mesothelioma growth. Histological analyses revealed that FG-3019 not only inhibited the proliferation but also induced apoptosis in both mesothelioma cells and fibroblasts. Our data suggest that FG-3019 antibody therapy could be a novel additional choice for the treatment of mesothelioma.


Transcriptome profiling of brown adipose tissue during cold exposure reveals extensive regulation of glucose metabolism.
We applied digital gene expression profiling to determine the transcriptome of brown and white adipose tissues (BAT and WAT, respectively) during cold exposure. Male C57BL/6J mice were exposed to cold for 2 or 4 days. A notable induction of genes related to glucose uptake, glycolysis, glycogen metabolism, and the pentose phosphate pathway was observed in BAT from cold-exposed animals. In addition, glycerol-3-phosphate dehydrogenase 1 expression was induced in BAT from cold-challenged mice, suggesting increased synthesis of glycerol from glucose. Similarly, expression of lactate dehydrogenases was induced by cold in BAT. Pyruvate dehydrogenase kinase 2 (Pdk2) and Pdk4 were expressed at significantly higher levels in BAT than in WAT, and Pdk2 was induced in BAT by cold. Of notice, only a subset of the changes detected in BAT was observed in WAT. Based on changes in gene expression during cold exposure, we propose a model for the intermediary glucose metabolism in activated BAT: 1) fluxes through glycolysis and the pentose phosphate pathway are induced, the latter providing reducing equivalents for de novo fatty acid synthesis; 2) glycerol synthesis from glucose is increased, facilitating triacylglycerol synthesis/fatty acid re-esterification; 3) glycogen turnover and lactate production are increased; and 4) entry of glucose carbon into the tricarboxylic acid cycle is restricted by PDK2 and PDK4. In summary, our results demonstrate extensive and diverse gene expression changes related to glucose handling in activated BAT.


Proteomic and functional analysis of proline dehydrogenase 1 link proline catabolism to mitochondrial electron transport in Arabidopsis thaliana.
Proline accumulates in many plant species in response to environmental stresses. Upon relief from stress, proline is rapidly oxidized in mitochondria by proline dehydrogenase (ProDH) and then by pyrroline-5-carboxylate dehydrogenase (P5CDH). Two ProDH genes have been identified in the genome of the model plant Arabidopsis thaliana To gain a better understanding of ProDH1 functions in mitochondria, proteomic analysis was performed. ProDH1 polypeptides were identified in Arabidopsis mitochondria by immunoblotting gels after 2D blue native (BN)-SDS/PAGE, probing them with an anti-ProDH antibody and analysing protein spots by MS. The 2D gels showed that ProDH1 forms part of a low-molecular-mass (70-140 kDa) complex in the mitochondrial membrane. To evaluate the contribution of each isoform to proline oxidation, mitochondria were isolated from wild-type (WT) and prodh1, prodh2, prodh1prodh2 and p5cdh mutants. ProDH activity was high for genotypes in which ProDH, most likely ProDH1, was strongly induced by proline. Respiratory measurements indicate that ProDH1 has a role in oxidizing excess proline and transferring electrons to the respiratory chain.


Post-thymic T lymphocyte maturation during ontogenesis.
Peripheral T lymphocytes from newborn (4-6-d-old) mice, isolated from the spleen or lymph nodes, show phenotypic features of immature cortical thymocytes, such as high frequencies of proliferating cells and of peanut lectin-binding cells. These are features of peripheral T cells of recent thymic origin, as shown by in situ labeling of thymocytes and subsequent observation of the migrants to the spleen, which were mainly peanut lectin-binding cells. The function of newborn peripheral T cells was compared, on a per T cell basis, with that of thymocytes and of fully mature peripheral T cells of the adult, using preparations of newborn lymph node cells containing approximately 80% of T lymphocytes. They were strikingly (about 10-fold) less competent than adult T cells in their phytohemagglutinin responsiveness, their capacities to induce a graft vs. host reaction, to proliferate in the mixed lymphocyte reaction, and to help B lymphocytes in a humoral response in vivo and in vitro. In contrast, newborn T lymphocytes were comparable to those of adults in their capacity to generate cytotoxic T lymphocytes. No suppressive effect of newborn T lymphocytes could be demonstrated in several of these assays. These results argue for an asynchronous maturation of two T cell subsets during ontogeny and demonstrate that at least some T lymphocytes leave the thymus as immature T cells resembling cortical thymocytes and further mature at the periphery. Investigation of mice submitted to thymectomy of 5 d of age showed that these incompetent post-thymic T lymphocytes are capable of considerable expansion and maturation in the peripheral lymphoid organs in the absence of a thymic influence.


Honey in Bronchial Asthma: From Folk Tales to Scientific Facts.
Bronchial asthma is one of the most common chronic inflammatory diseases. Complementary and alternative medicine is increasingly used for treating bronchial asthma. Ten electronic databases were searched to investigate whether honey alone or in combination with other ingredients can be considered as the potential treatment for bronchial asthma. Combinations of honey and Nigella sativa (NS) showed significant improvement in all pulmonary functions, including forced expiratory volume (FEV1) (MD = 0.52, P < .001), forced vital capacity (FVC) (MD = 0.55, P = .002), and peak expiratory flow rate (PEFR) (MD = 80.60, P < .001), in both moderate and severe, uncontrolled persistent asthma compared with baseline. Asthma control test scores also improved significantly (MD = 11.22, P < .001) in patients using combinations of honey and NS compared with baseline. Patients with a less severe grade of asthma showed a significant positive response in clinical parameters upon using honey. One study showed that using celery seeds and honey was associated with clinical improvement of both lung functions, FEV1 (MD = 18.09, P < .001) and FVC (MD = 24.23, P < .001), and respiratory parameters compared with baseline. In conclusion, honey alone has no strong evidence of being effective in controlling asthma. However, when used in combination with other substances, it showed a relatively high efficacy in patients with asthma. This finding may help in asthma control with lower cost alternatives and better outcomes.


Losing Neutrality: The Neural Basis of Impaired Emotional Control without Sleep.
Sleep deprivation has been shown recently to alter emotional processing possibly associated with reduced frontal regulation. Such impairments can ultimately fail adaptive attempts to regulate emotional processing (also known as cognitive control of emotion), although this hypothesis has not been examined directly. Therefore, we explored the influence of sleep deprivation on the human brain using two different cognitive-emotional tasks, recorded using fMRI and EEG. Both tasks involved irrelevant emotional and neutral distractors presented during a competing cognitive challenge, thus creating a continuous demand for regulating emotional processing. Results reveal that, although participants showed enhanced limbic and electrophysiological reactions to emotional distractors regardless of their sleep state, they were specifically unable to ignore neutral distracting information after sleep deprivation. As a consequence, sleep deprivation resulted in similar processing of neutral and negative distractors, thus disabling accurate emotional discrimination. As expected, these findings were further associated with a decrease in prefrontal connectivity patterns in both EEG and fMRI signals, reflecting a profound decline in cognitive control of emotion. Notably, such a decline was associated with lower REM sleep amounts, supporting a role for REM sleep in overnight emotional processing. Altogether, our findings suggest that losing sleep alters emotional reactivity by lowering the threshold for emotional activation, leading to a maladaptive loss of emotional neutrality. Significance statement: Sleep loss is known as a robust modulator of emotional reactivity, leading to increased anxiety and stress elicited by seemingly minor triggers. In this work, we aimed to portray the neural basis of these emotional impairments and their possible association with frontal regulation of emotional processing, also known as cognitive control of emotion. Using specifically suited EEG and fMRI tasks, we were able to show that sleep deprivation alters emotional reactivity by triggering enhanced processing of stimuli regarded previously as neutral. These changes were further accompanied by diminished frontal connectivity, reduced REM sleep, and poorer performance. Therefore, we suggest that sleep loss alters emotional reactivity by lowering the threshold for emotional activation, leading to a maladaptive loss of emotional neutrality.


Population variation of the fall armyworm, Spodoptera frugiperda, in the Western Hemisphere.
Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), the fall armyworm is the most economically important maize pest in the western hemisphere. This research focused on the genetic variability of the maize host strain because there is a lack of information in this area of S. frugiperda research. Amplified fragment length polymorphism (AFLP) was used to assess the genetic variability of S. frugiperda over a large geographic area. Twenty populations were collected from the maize, one population was collected from princess tree, one population was collected from lemon tree, and one population was collected from bermudagrass. The 23 populations were from Mexico, the continental United States, Puerto Rico, Brazil, and Argentina. The objective of this research was to evaluate whether the majority of genetic variability was within populations or between populations. The AFLP results showed that the majority of the genetic variability is within populations and not between populations, indicating minor gene flow and suggesting that S. frugiperda in the Western Hemisphere are an interbreeding population.


Joy of life in nursing homes: A qualitative study of what constitutes the essence of Joy of life in elderly individuals living in Norwegian nursing homes.
Finding new approaches to increase positive health and well-being of elderly individuals living in nursing homes (NH) is highly warranted. Several Norwegian municipalities have implemented the certification scheme framed 'Joy of Life Nursing Home' (JOLNH). JOLNH is based on a health-promoting perspective directing elderly individuals' resources. Through health promotion, preventive and social activities across generations, NH care promotes respect, well-being, health and cultural experiences among elderly individuals. The aims of this study were to explore the phenomenon of Joy of life (JOL) among elderly individuals living in NHs and to provide a deeper understanding of which dimensions constitute JOL in this population. The present study explored the JOL phenomenon using qualitative methodology. Data were collected between December 2015 and May 2016 by individual qualitative research interviews of 29 elderly individuals who met the inclusion criteria. The informants were living in ten different NHs in two large municipalities in Norway. Prior approval was obtained from all regulatory institutions dealing with research issues and the management in both municipalities. The results showed that (i) positive relations, (ii) a sense of belonging, (iii) sources of meaning, (iv) moments of feeling well, and (v) acceptance constituted the conceptualisation of the JOL phenomenon. The dimensions constituting the JOL concept correspond to the concepts of well-being and flourishing. Knowledge of the JOL phenomenon may help professionals to target elderly individuals' health and well-being more precisely and thereby increase flourishing, JOL and well-being among them.


Identification and profiling of miRNAs during herbivory reveals jasmonate-dependent and -independent patterns of accumulation in Nicotiana attenuata.
Plant microRNAs (miRNAs) play key roles in the transcriptional responses to environmental stresses. However, the role of miRNAs in responses to insect herbivory has not been thoroughly explored. To identify herbivory-responsive miRNAs, we identified conserved miRNAs in the ecological model plant Nicotiana attenuata whose interactions with herbivores have been well-characterized in both laboratory and field studies. We identified 59 miRNAs from 36 families, and two endogenous trans-acting small interfering RNAs (tasiRNA) targeted by miRNAs. We characterized the response of the precursor and mature miRNAs to simulated attack from the specialist herbivore Manduca sexta by quantitative PCR analysis and used ir-aoc RNAi transformants, deficient in jasmonate biosynthesis, to identify jasmonate-dependent and -independent miRNA regulation. Expression analysis revealed that groups of miRNAs and tasiRNAs were specifically regulated by either mechanical wounding or wounding plus oral secretions from M. sexta larvae, and these small RNAs were accumulated in jasmonate-dependent or -independent manners. Moreover, cDNA microarray analysis indicated that the expression patterns of the corresponding target genes were correlated with the accumulation of miRNAs and tasiRNAs. We show that a group of miRNAs and tasiRNAs orchestrates the expression of target genes involved in N. attenuata's responses to herbivore attack.


Type 1 and 2 immunity following vaccination is influenced by nanoparticle size: formulation of a model vaccine for respiratory syncytial virus.
Previous studies compared uptake by dendritic cells (DC) of 20, 40, 100, 200, 500, 1000, and 2000 nm beads in vivo. When beads were used as antigen carriers, bead size influenced antibody responses and induction of IFN-gamma-producing CD4 and CD8 T cells. Beads of 40-50 nm were taken up preferentially by DC and induced particularly strong immunity. Herein, we examine immunity induced by minute differences in nanobead size, specifically within a narrow viral-sized range (20, 40, 49, 67, 93, 101, and 123 nm), to see if bead carrier size influenced the induction of type 1 or type 2 cells as demonstrated by the production of IFN-gamma or IL-4. In vivo uptake by DC was assessed for selected sizes in this range. Responses to whole ovalbumin (OVA) or the OVA-derived CD8 T cell peptide epitope (SIINFEKL) were tested. After one immunization with beads-OVA, IFN-gamma responses to both OVA and SIINFEKL were significantly better with 40 and 49 nm beads than other sizes, while, in contrast, IL-4 responses to OVA were higher after immunization with OVA conjugated to larger beads (93, 101, and 123 nm). Thus IFN-gamma induction from CD8 T cells was limited to 40-49 nm beads, while CD4 T cell activation and IL-4 were induced by 93-123 nm beads-OVA. After two immunizations, there were comparable high levels of IFN-gamma produced with 40 and 49 beads and IL-4 reactivity was still higher for larger beads (93, 101, 123 nm). Production of IgG1 was seen across the full range of bead sizes, increasing after two immunizations. Since protection against respiratory syncytial virus (RSV) depends on strong IFN responses, while IL-4 responses are reported to cause asthma-like symptoms, immunization with RSV antigens on the 49 nm carrier beads could provide the basis for a suitable vaccine. When the 49 nm beads were conjugated to RSV proteins G88 (surface) or M2.1 (internal capsid), one immunization with G88 induced high levels of IFN-gamma and low levels of IL-4. IL-4 increased with two immunizations. Beads-M2.1 induced only moderate levels of IFN-gamma and low titer antibody after two immunizations. Mice vaccinated once with G88-conjugated 49 nm beads and challenged intranasally with RSV strain A2 subtype showed reduced viral titers and recovered from weight loss more rapidly than mice immunized with M2.1-conjugated 49 nm beads or naive control mice. These results show that precise selection of nanobead size for vaccination can influence the type 1/type 2 cytokine balance after one immunization, and this will be useful in the development of effective vaccines against common human pathogens such as RSV.


Evaluation of GERD symptoms during therapy. Part II. Psychometric evaluation and validation of the new questionnaire ReQuest in erosive GERD.
Evaluation of the response of gastroesophageal reflux disease (GERD) symptoms to treatment would be facilitated by a brief, valid, reliable and responsive, self-assessed GERD-sensitive scale. We therefore developed the Reflux Questionnaire (ReQuest). This publication describes the psychometric evaluation and validation of ReQuest. This second phase of development was based on data from a clinical trial of patients with erosive GERD who received pantoprazole 20 or 40 mg daily for 28 days and completed weekly the long, and daily the short version of ReQuest. The psychometric analyses of ReQuest included internal consistency, test-retest reliability and responsiveness. Construct validity was evaluated by comparison with the Gastrointestinal Symptom Rating Scale (GSRS) and the Psychological General Well-Being (PGWB) scale. Validation of ReQuest indicated very high internal consistency (Cronbach's alpha = 0.90) and test-retest reliability (intraclass correlation coefficient 0.94 (long-long) and 0.86 (short-short)). This was also the case for the two subscales ReQuest-GI and ReQuest-WSO with Cronbach's alpha coefficients of 0.84 and 0.81. Responsiveness was high with a responsiveness index of >0.8 at day 28. Construct validity was good. ReQuest is a highly reliable, valid and responsive self-assessment tool for evaluating treatment response in patients with erosive GERD, and can be applied daily.


Macular Vascular Imaging and Connectivity Analysis Using High-Resolution Optical Coherence Tomography.
To characterize macular blood flow connectivity in vivo using high-resolution optical coherence tomography (HighRes OCT). Cross-sectional, observational study. Dense (6-µm interscan distance) perifoveal HighRes OCT raster scans were performed on healthy participants. To mitigate the limitations of projection-resolved OCT-angiography, flow and structural data were used to observe the vascular structures of the superficial vascular complex (SVC) and the deep vascular complex. Vascular segmentation and rendering were performed using Imaris 9.5 software. Inflow and outflow patterns were classified according to vascular diameter and branching order from superficial arteries and veins, respectively. Eight eyes from eight participants were included in this analysis, from which 422 inflow and 459 outflow connections were characterized. Arteries had direct arteriolar connections to the SVC (78%) and to the intermediate capillary plexus (ICP, 22%). Deep capillary plexus (DCP) inflow derived from small-diameter vessels succeeding ICP arterioles. The most prevalent outflow pathways coursed through superficial draining venules (74%). DCP draining venules ordinarily merged with ICP draining venules and drained independently of superficial venules in 21% of cases. The morphology of DCP draining venules in structural HighRes OCT is distinct from other vessels crossing the inner nuclear layer and can be used to identify superficial veins. Vascular connectivity analysis supports a hybrid circuitry of blood flow within the human parafoveal macula. Characterization of parafoveal macular blood flow connectivity in vivo using a precise segmentation of HighRes OCT is consistent with ground-truth microscopy studies and shows a hybrid circuitry.


Prophylaxis of implant-related infections by local release of vancomycin from a hydrogel in rabbits.
Local prophylaxis with antibiotic-loaded bone cement is a successful method to prevent post-operative infections in patients receiving orthopaedic implants. No comparable method is available for uncemented implants. Therefore, a hydrogel consisting of hyaluronic and polylactic acids was evaluated in a rabbit model for delivery of antimicrobial agents to prevent post-operative infections. In a pilot study, the suitability of the in vivo model was assessed by testing the hydrogel as carrier material for antimicrobial agents.In the main study, the antimicrobial-agent-loaded hydrogel was evaluated for infection prophylaxis. Rabbits received a titanium rod intramedullary in the tibia after contamination with Staphylococcus aureus. The rods were coated with unloaded hydrogel (Gel), hydrogel loaded with 2 % (Van2) or 5 % vancomycin (Van5), bioactive glass (BAG) or N-acetyl-L-cysteine (NAC). To analyse the infection severity after 28 d, histopathological, bacteriological, micro-computed tomographic and haematological analyses were performed. In the pilot study, the Van5 group had less infection (0/6 infected) as compared to the Gel group (5/5, p = 0.000) and the in vivo model was deemed suitable. In the main study, in the Van2 and Van5 groups, the number of infected animals was lower [1/6 (p = 0.006) and 2/6 (p = 0.044) infected, respectively]. In contrast, BAG and NAC groups showed no infection reduction (5/6 both groups, p = 0.997). The hydrogel can be used as a local carrier of vancomycin for prophylaxis of implant-related infections.The present study showed promising results for local delivery of antibacterial agents by hydrogel to prevent implant-related infections.


Chronic alcohol self-administration in monkeys shows long-term quantity/frequency categorical stability.
The current criteria for alcohol use disorders (AUDs) do not include consumption (quantity/frequency) measures of alcohol intake, in part due to the difficulty of these measures in humans. Animal models of ethanol (EtOH) self-administration have been fundamental in advancing our understanding of the neurobiological basis of AUD and can address quantity/frequency measures with accurate measurements over prolonged periods of time. The nonhuman primate model of voluntary oral alcohol self-administration has documented both binge drinking and drinking to dependence and can be used to test the stability of consumption measures over time. Here, an extensive set of alcohol intakes (g/kg/d) was analyzed from a large multi-cohort population of Rhesus (Macaca mulatta) monkeys (n = 31). Daily EtOH intake was uniformly distributed over chronic (12 months) access for all animals. Underlying this distribution of intakes were subpopulations of monkeys that exhibited distinctive clustering of drinking patterns, allowing us to categorically define very heavy drinking (VHD), heavy drinking (HD), binge drinking (BD), and low drinking (LD). These categories were stable across the 12 months assessed by the protocol, but exhibited fluctuations when examined at shorter intervals. The establishment of persistent drinking categories based on quantity/frequency suggests that consumption variables can be used to track long-term changes in behavioral, molecular, or physiochemical mechanisms related to our understanding of diagnosis, prevention, intervention, and treatment efficacies.


GRK2-mediated inhibition of adrenergic and dopaminergic signaling in right ventricular hypertrophy: therapeutic implications in pulmonary hypertension.
The cause and consequences of impaired adrenergic signaling in right ventricular failure/hypertrophy (RVH) are poorly understood. We hypothesized that G protein-coupled receptor kinase-2 (GRK2)-mediated uncoupling of β-adrenergic receptor signaling impairs inotropic reserve. The implications of right ventricular (RV) adrenergic remodeling for inotrope selection and the therapeutic benefit of interrupting Gβγ-GRK2 interaction, using gallein, were tested. Chamber-specificity and cellular localization of adrenergic remodeling were compared in rodent RVH associated with pulmonary arterial hypertension (PAH-RVH; SU5416+chronic-hypoxia or Monocrotaline) versus pulmonary artery banding-induced RVH (PAB-RVH). Results were corroborated in RV arrays from 10 PAH patients versus controls. Inotropic reserve was assessed in RV- and left ventricular-Langendorff models and in vivo. Gallein therapy (1.8 mg/kg/day ×2-weeks) was assessed. Despite similar RVH, cardiac output (58.3±4.9 versus 82.9±4.8 mL/min; P<0.001) and treadmill distance (41.5±11.6 versus 244.1±12.4 m; P<0.001) were lower in PAH-RVH versus PAB-RVH. In PAH-RVH versus PAB-RVH there was greater downregulation of β1-, α1- and dopamine-1 receptors, more left ventricular involvement, and greater impairment of RV contractile reserve. RV GRK2 activity increased in parallel with a reduction in both adrenergic receptor expression and inotrope-stimulated cAMP levels (P<0.01). β1-receptor downregulation also occurred in human PAH-RVH. Dobutamine was superior to dopamine as an RV inotrope, both ex vivo and in vivo. GRK2-mediated desensitization-downregulation of adrenergic and dopaminergic receptors impairs inotropic reserve in PAH-RVH. Acute inotropic support in RVH is best accomplished by dobutamine, reflecting its better coupling to adenylyl cyclase and the reliance of dopamine on dopamine-1-receptor signaling, which is impaired in RVH. Inhibiting Gβγ-GRK2 interactions has therapeutic benefit in RVH.


Local versus intravenous injections of skeletal muscle precursor cells in nonhuman primates with acute or chronic intrinsic urinary sphincter deficiency.
Many factors may influence the efficacy of cell therapy for intrinsic urinary sphincter deficiency (ISD), including the route of administration of the cells and the condition of the sphincter. The goal of this study was to compare local versus intravenous administration of autologous skeletal muscle precursor cells (skMPCs) when administered to nonhuman primates (NHPs) with either acute or chronic ISD. Thirty-two adult female monkeys were divided into eight groups (n = 4/group): (1) control; (2) surgically induced ISD/no treatment; (3) acute ISD (6-week duration)/local vehicle only; (4) acute ISD/local skMPC injection; (5) acute ISD/systemic skMPC; (6) chronic ISD (6-month duration)/local vehicle; (7) chronic ISD/local skMPC; (8) chronic ISD/systemic skMPC. Maximal urethral pressures (MUP) were measured prior to ISD, prior to treatment and at 3 and 6 months following treatment. Quantitative histology was used to measure muscle/collagen content, somatic innervation, and vascularity of the sphincter complexes. In NHPs with acute ISD both systemic and local administration of skMPCs increased resting MUP values and sphincter muscle content (p < 0.05 vs. ISD/vehicle). However, the effects of systemic skMPC administration were significantly lower than those of local injection (p > 0.05). In NHPs with chronic ISD local skMPC administration had reduced (compared to NHPs with acute ISD) effects on MUP and sphincter muscle values (p < 0.05 vs. acute ISD/skMPC); systemic administration had no effect. Pudendal nerve-stimulated increases in MUP were significant only in acute ISD NHPs with local skMPC treatment (p < 0.05 vs. resting MUP). The extent of sphincter vascularization and innervation were directly related to MUP and sphincter muscle content. Both the chronicity of ISD and the route of cell injection influence the efficacy of cell therapy in monkey models of ISD. This may be related to the relative ability of cells to stimulate vascularization and re-innervation in these different treatment conditions.


The SARS-CoV-2 Ivermectin Navarra-ISGlobal Trial (SAINT) to Evaluate the Potential of Ivermectin to Reduce COVID-19 Transmission in low risk, non-severe COVID-19 patients in the first 48 hours after symptoms onset: A structured summary of a study protocol for a randomized control pilot trial.
The primary objective is to determine the efficacy of a single dose of ivermectin, administered to low risk, non-severe COVID-19 patients in the first 48 hours after symptom onset to reduce the proportion of patients with detectable SARS-CoV-2 RNA by Polymerase Chain Reaction (PCR) test from nasopharyngeal swab at day 7 post-treatment. The secondary objectives are: 1.To assess the efficacy of ivermectin to reduce the SARS-CoV-2 viral load in the nasopharyngeal swab at day 7 post treatment.2.To assess the efficacy of ivermectin to improve symptom progression in treated patients.3.To assess the proportion of seroconversions in treated patients at day 21.4.To assess the safety of ivermectin at the proposed dose.5.To determine the magnitude of immune response against SARS-CoV-2.6.To assess the early kinetics of immunity against SARS-CoV-2. SAINT is a single centre, double-blind, randomized, placebo-controlled, superiority trial with two parallel arms. Participants will be randomized to receive a single dose of 400 μg/kg ivermectin or placebo, and the number of patients in the treatment and placebo groups will be the same (1:1 ratio). The population for the study will be patients with a positive nasopharyngeal swab PCR test for SARS-CoV-2, with non-severe COVID-19 disease, and no risk factors for progression to severity. Vulnerable populations such as pregnant women, minors (i.e.; under 18 years old), and seniors (i.e.; over 60 years old) will be excluded. Inclusion criteria 1. Patients diagnosed with COVID-19 in the emergency room of the Clínica Universidad de Navarra (CUN) with a positive SARS-CoV-2 PCR. 2. Residents of the Pamplona basin ("Cuenca de Pamplona"). 3. The patient must be between the ages of 18 and 60 years of age. 4. Negative pregnancy test for women of child bearing age*. 5. The patient or his/her representative, has given informed consent to participate in the study. 6. The patient should, in the PI's opinion, be able to comply with all the requirements of the clinical trial (including home follow up during isolation). Exclusion criteria 1. Known history of ivermectin allergy. 2. Hypersensitivity to any component of ivermectin. 3. COVID-19 pneumonia. Diagnosed by the attending physician.Identified in a chest X-ray. 4. Fever or cough present for more than 48 hours. 5. Positive IgG against SARS-CoV-2 by rapid diagnostic test. 6. Age under 18 or over 60 years. 7. The following co-morbidities (or any other disease that might interfere with the study in the eyes of the PI): Immunosuppression.Chronic Obstructive Pulmonary Disease.Diabetes.Hypertension.Obesity.Acute or chronic renal failure.History of coronary disease.History of cerebrovascular disease.Current neoplasm. 8. Recent travel history to countries that are endemic for Loa loa (Angola, Cameroon, Central African Republic, Chad, Democratic Republic of Congo, Ethiopia, Equatorial, Guinea, Gabon, Republic of Congo, Nigeria and Sudan). 9. Current use of CYP 3A4 or P-gp inhibitor drugs such as quinidine, amiodarone, diltiazem, spironolactone, verapamil, clarithromycin, erythromycin, itraconazole, ketoconazole, cyclosporine, tacrolimus, indinavir, ritonavir or cobicistat. Use of critical CYP3A4 substrate drugs such as warfarin. *Women of child bearing age may participate if they use a safe contraceptive method for the entire period of the study and at least one month afterwards. A woman is considered to not have childbearing capacity if she is post-menopausal (minimum of 2 years without menstruation) or has undergone surgical sterilization (at least one month before the study). The trial is currently planned at a single center, Clínica Universidad de Navarra, in Navarra (Spain), and the immunology samples will be analyzed at the Barcelona Institute for Global Health (ISGlobal), in Barcelona (Spain). Participants will be recruited by the investigators at the emergency room and/or COVID-19 area of the CUN. They will remain in the trial for a period of 28 days at their homes since they will be patients with mild disease. In the interest of public health and to contain transmission of infection, follow-up visits will be conducted in the participant's home by a clinical trial team comprising nursing and medical members. Home visits will assess clinical and laboratory parameters of the patients. Ivermectin will be administered to the treatment group at a 400μg/Kg dose (included in the EU approved label of Stromectol and Scabioral). The control group will receive placebo. There is no current data on the efficacy of ivermectin against the virus in vivo, therefore the use of placebo in the control group is ethically justified. Primary Proportion of patients with a positive SARS-CoV-2 PCR from a nasopharyngeal swab at day 7 post-treatment. Secondary 1.Mean viral load as determined by PCR cycle threshold (Ct) at baseline and on days 4, 7, 14, and 21.2.Proportion of patients with fever and cough at days 4, 7, 14, and 21 as well as proportion of patients progressing to severe disease or death during the trial.3.Proportion of patients with seroconversion at day 21.4.Proportion of drug-related adverse events during the trial.5.Median levels of IgG, IgM, IgA measured by Luminex, frequencies of innate and SARS-CoV-2-specific T cells assessed by flow cytometry, median levels of inflammatory and activation markers measured by Luminex and transcriptomics.6.Median kinetics of IgG, IgM, IgA levels during the trial, until day 28. Eligible patients will be allocated in a 1:1 ratio using a randomization list generated by the trial statistician using blocks of four to ensure balance between the groups. A study identification code with the format "SAINT-##" (##: from 01 to 24) will be generated using a sequence of random numbers so that the randomization number does not match the subject identifier. The sequence and code used will be kept in an encrypted file accessible only to the trial statistician. A physical copy will be kept in a locked cabinet at the CUN, accessible only to the person administering the drug who will not enrol or attend to patient care. A separate set of 24 envelopes for emergency unblinding will be kept in the study file. The clinical trial team and the patients will be blinded. The placebo will not be visibly identical, but it will be administered by staff not involved in the clinical care or participant follow up. The sample size is 24 patients: 12 participants will be randomised to the treatment group and 12 participants to the control group. Current protocol version: 1.0 dated 16 of April 2020. Recruitment is envisioned to begin by May 14th and end by June 14th. EudraCT number: 2020-001474-29, registered April 1st. Clinicaltrials.gov: submitted, pending number FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Effect of cortisol on the immune-like response of rainbow trout (Oncorhynchus mykiss) myotubes challenged with Piscirickettsia salmonis.
Salmonids are a species of high commercial value in Chilean aquaculture, where muscle is the final product of the industry. Fish can be affected by stress during intensive cultures, increasing susceptibility to infections. Recently, we reported that muscle is an important focus of immune reactions. However, studies have shown the immunosuppressive effect of stress only in lymphoid organs, and few studies have been conducted on muscle and immunity. Hence, we determine the effects of cortisol on the immune-like response of fish myotubes challenged with Piscirickettsia salmonis by three trials. First, rainbow trout primary culture of muscle was cultured and treated with cortisol (100 ng/mL) for 3 and 4 h. Second, myotubes were challenged with P. salmonis (MOI 50) for 4, 6 and 8 h. And third, muscle cell cultures were pretreated with cortisol and then challenged with P. salmonis. The mRNA levels of glucocorticoid pathway and innate immunity were evaluated by qPCR. Cortisol increased the klf15 levels and downregulated the innate immune-related tlr5m gene and antimicrobial peptides. P. salmonis challenge upregulated several immune-related genes. Finally, cortisol pretreatment followed by P. salmonis challenge differentially modulated stress- and immune-related genes. These data suggest that fish muscle cells possess an intrinsic immune response and are differentially regulated by cortisol, which could lead to bacterial outbreaks in muscle under stress conditions.


Faithful reconstruction of digital holograms captured by FINCH using a Hamming window function in the Fresnel propagation.
Recent advances in Fresnel incoherent correlation holography (FINCH) increase the signal-to-noise ratio in hologram recording by interference of images from two diffractive lenses with focal lengths close to the image plane. Holograms requiring short reconstruction distances are created that reconstruct poorly with existing Fresnel propagation methods. Here we show a dramatic improvement in reconstructed fluorescent images when a 2D Hamming window function substituted for the disk window typically used to bound the impulse response in the Fresnel propagation. Greatly improved image contrast and quality are shown for simulated and experimentally determined FINCH holograms using a 2D Hamming window without significant loss in lateral or axial resolution.


Copper supplementation has no effect on markers of DNA damage and liver function in healthy adults (FOODCUE project).
Copper is routinely used in the laboratory to promote oxidation in vitro. However, copper concentrations are million-fold higher than physiological concentrations and, in contrast, accumulating evidence suggests that copper may have an antioxidant role in vivo. The aim of this study was to provide data on how increased intake of copper affected mononuclear leukocyte DNA damage and liver function in healthy young free-living men and women. The study design was a double-blind repeated crossover trial with treatment and intervening placebo periods, each of 6 weeks' duration. The following supplementations were given orally in sequence: CuSO(4) at a dose of 3 mg copper/day and copper amino acid chelates at doses of 3 and 6 mg copper/day. Oxidative DNA damage was assessed using a modification of the alkaline Comet assay incorporating an endonuclease III digestion step. The assessment of liver function was by measurement of the liver enzymes, alanine aminotransferase and L-gamma-glutamyltransferase. There was no significant alteration in mononuclear leukocyte DNA damage or on liver function after 6 weeks of copper supplementation at two doses (3 and 6 mg/day). Copper supplementation (giving total copper intake at the highest level of 7 mg/day) did not induce DNA damage or adversely affect liver function in healthy adults.


Production Assistance for Cellular Therapies (PACT): four-year experience from the United States National Heart, Lung, and Blood Institute (NHLBI) contract research program in cell and tissue therapies.
In 2002, the US National Heart, Lung, and Blood Institute (NHLBI) conducted a workshop to determine needs of the cell therapy community. A consensus emerged that improved access to cGMP facilities, regulatory assistance, and training would foster the advancement of cellular therapy. A 2003 NHLBI request for proposals resulted in four contracts being awarded to three cell-manufacturing facilities (Baylor College of Medicine, University of Minnesota, and University of Pittsburgh) and one administrative center (The EMMES Corporation). As a result, Production Assistance for Cellular Therapies (PACT) was formed. As of October 1, 2008, PACT has received 65 preliminary applications of which 45 have been approved for product manufacture. A variety of cell therapies are represented including T-regulatory cells, natural killer cells, adipose-derived stem cells, cardiac progenitor cells for cardiac disease, hematopoietic progenitor cells (HPCs) for central nervous system applications, cytotoxic T lymphocytes, and dendritic cells. A total of 169 products have been administered under 12 applications and 2 reagents were manufactured and delivered. Fourteen peer-reviewed publications and 15 abstracts have resulted from the PACT project to date. A cell therapy textbook is nearly complete. PACT technical projects have addressed assay development, rapid endotoxin testing, shipping of cell products, and CD34+ HPC isolation from low-volume marrow. Educational Web seminars and on-site training through workshops have been conducted. PACT is an active and successful cell therapy manufacturing resource in the United States, addressing research and training while forging relationships among academia, industry, and participating institutions.


Effect of physiological age on radiation resistance of some bacteria that are highly radiation resistant.
Physiological age-dependent variation in radiation resistance was studied for three bacteria that are highly radiation resistant: Micrococcus radiodurans, Micrococcus sp. isolate C-3, and Moraxella sp. isolate 4. Stationary-phase cultures of M. radiodurans and isolate C-3 were much more resistant to gamma radiation than were log-phase cultures. This pattern of relative resistance was reversed for isolate 4. Resistance of isolate 4 to UV light was also greater during log phase, although heat resistance and NaCl tolerance after heat stress were greater during stationary phase. Radiation-induced injury of isolate 4 compared with injury of Escherichia coli B suggested that the injury process, as well as the lethal process, was affected by growth phase. The hypothesis that growth rate affects radiation resistance was tested, and results were interpreted in light of the probable confounding effect of methods used to alter growth rates of bacteria. These results indicate that dose-response experiments should be designed to measure survival during the most resistant growth phase of the organism under study. This timing is particularly important when extrapolations of survival results might be made to potential irradiation processes for foods.


Genome-wide association study of dietary intake in the UK biobank study and its associations with schizophrenia and other traits.
Motivated by observational studies that report associations between schizophrenia and traits, such as poor diet, increased body mass index and metabolic disease, we investigated the genetic contribution to dietary intake in a sample of 335,576 individuals from the UK Biobank study. A principal component analysis applied to diet question item responses generated two components: Diet Component 1 (DC1) represented a meat-related diet and Diet Component 2 (DC2) a fish and plant-related diet. Genome-wide association analysis identified 29 independent single-nucleotide polymorphisms (SNPs) associated with DC1 and 63 SNPs with DC2. Estimated from over 35,000 3rd-degree relative pairs that are unlikely to share close family environments, heritabilities for both DC1 and DC2 were 0.16 (standard error (s.e.) = 0.05). SNP-based heritability was 0.06 (s.e. = 0.003) for DC1 and 0.08 (s.e = 0.004) for DC2. We estimated significant genetic correlations between both DCs and schizophrenia, and several other traits. Mendelian randomisation analyses indicated a negative uni-directional relationship between liability to schizophrenia and tendency towards selecting a meat-based diet (which could be direct or via unidentified correlated variables), but a bi-directional relationship between liability to schizophrenia and tendency towards selecting a fish and plant-based diet consistent with genetic pleiotropy.


Risk Distribution of Human Infections with Avian Influenza H7N9 and H5N1 virus in China.
It has been documented that the epidemiological characteristics of human infections with H7N9 differ significantly between H5N1. However, potential factors that may explain the different spatial distributions remain unexplored. We use boosted regression tree (BRT) models to explore the association of agro-ecological, environmental and meteorological variables with the occurrence of human cases of H7N9 and H5N1, and map the probabilities of occurrence of human cases. Live poultry markets, density of human, coverage of built-up land, relative humidity and precipitation were significant predictors for both. In addition, density of poultry, coverage of shrub and temperature played important roles for human H7N9 infection, whereas human H5N1 infection was associated with coverage of forest and water body. Based on the risks and distribution of ecological characteristics which may facilitate the circulation of the two viruses, we found Yangtze River Delta and Pearl River Delta, along with a few spots on the southeast coastline, to be the high risk areas for H7N9 and H5N1. Additional, H5N1 risk spots were identified in eastern Sichuan and southern Yunnan Provinces. Surveillance of the two viruses needs to be enhanced in these high risk areas to reduce the risk of future epidemics of avian influenza in China.


Potential role of seaweeds in climate change mitigation.
Seaweed (macroalgae) has attracted attention globally given its potential for climate change mitigation. A topical and contentious question is: Can seaweeds' contribution to climate change mitigation be enhanced at globally meaningful scales? Here, we provide an overview of the pressing research needs surrounding the potential role of seaweed in climate change mitigation and current scientific consensus via eight key research challenges. There are four categories where seaweed has been suggested to be used for climate change mitigation: 1) protecting and restoring wild seaweed forests with potential climate change mitigation co-benefits; 2) expanding sustainable nearshore seaweed aquaculture with potential climate change mitigation co-benefits; 3) offsetting industrial CO2 emissions using seaweed products for emission abatement; and 4) sinking seaweed into the deep sea to sequester CO2. Uncertainties remain about quantification of the net impact of carbon export from seaweed restoration and seaweed farming sites on atmospheric CO2. Evidence suggests that nearshore seaweed farming contributes to carbon storage in sediments below farm sites, but how scalable is this process? Products from seaweed aquaculture, such as the livestock methane-reducing seaweed Asparagopsis or low carbon food resources show promise for climate change mitigation, yet the carbon footprint and emission abatement potential remains unquantified for most seaweed products. Similarly, purposely cultivating then sinking seaweed biomass in the open ocean raises ecological concerns and the climate change mitigation potential of this concept is poorly constrained. Improving the tracing of seaweed carbon export to ocean sinks is a critical step in seaweed carbon accounting. Despite carbon accounting uncertainties, seaweed provides many other ecosystem services that justify conservation and restoration and the uptake of seaweed aquaculture will contribute to the United Nations Sustainable Development Goals. However, we caution that verified seaweed carbon accounting and associated sustainability thresholds are needed before large-scale investment into climate change mitigation from seaweed projects.


Case management protocol and declining blood lead concentrations among children.
Blood lead concentrations among children aged 6 years and younger become a concern at 10 microg/dL (0.48 micromol/L) or higher. The authors' objective was to determine whether initial blood lead concentrations of 10-19 microg/dL (0.48-0.96 micromol/L) declined among children aged 3 years and younger and whether the magnitude of decline was associated with the case management protocol of the state or local childhood lead poisoning prevention program. The authors analyzed childhood blood lead surveillance data from 1994 through 1995 and case management protocols from six states that reported the results of all blood lead tests. The study included 2109 children aged 2 years or younger who had a venous blood lead concentration of 10-19 microg/dL (0.48-0.96 micromol/L) and a follow-up venous blood lead test within 3 to 12 months. Overall, blood lead concentrations increased by 0.25 microg/dL (0.01 micromol/L) between the time of the initial elevated blood lead test and the follow-up test, but concentrations declined by 1.96 microg/dL (0.09 micromol/L) among children covered by a case management protocol that included a home visit and by 0.92 microg/dL (0.04 micromol/L) among those covered by a protocol that included a lead source investigation. The decline remained significant after we adjusted for the child's age. These findings suggest that childhood lead prevention programs should consider focusing their efforts on home visits and lead source investigations.


Quantitative imaging in breast tomosynthesis and CT: comparison of detection and estimation task performance.
This work investigates a framework for modeling volumetric breast imaging to compare detection and estimation task performance and optimize quantitative breast imaging. Volumetric reconstructions of a breast phantom, which incorporated electronic, quantum, and anatomical noise with embedded spherical lesions, were simulated over a range of acquisition angles varying from 4 degrees to 204 degrees with a constant total acquisition dose of 1.5 mGy. A maximum likelihood estimator was derived in terms of the noise power spectrum, which yielded figures of merit for quantitative imaging performance in terms of accuracy and precision. These metrics were computed for estimation of lesion area, volume, and location. Estimation task performance was optimized as a function of acquisition angle and compared to the performance of a more conventional lesion detection task. Results revealed tradeoffs between electronic, quantum, and anatomical noise. The detection of a 4 mm sphere was optimal at an acquisition angle of 84 degrees, where reconstructed images using a smaller acquisition angle exhibited increased anatomical noise and reconstructed images using a larger acquisition angle exhibited increased quantum and electronic noise. For all estimation tasks, accuracy was found to be fairly constant as a function acquisition angle indicating adequate system calibration, whereas a more significant dependence on acquisition angle was observed for precision performance. Precision for the 2D area estimation task was optimal at approximately 104 degrees, while precision of the 3D volume estimation task was optimal at larger angles (approximately 124 degrees). Precision for the localization task showed orientation dependence where localization was significantly inferior in the depth direction. Overall, precision for localization was optimal at larger angles (i.e., > 125 degrees) compared to the size estimation tasks. Results suggested that for quantitative imaging tasks, the acquisition angle should be larger than currently used in conventional breast tomosynthesis for lesion detection. Analysis of quantitative imaging performance using Fourier-based metrics highlights the difference between estimation and detection task in volumetric breast imaging and provides a meaningful framework for optimizing the performance of breast imaging systems for quantitative imaging applications.


The role of allostery in the ubiquitin-proteasome system.
The ubiquitin-proteasome system (UPS) is involved in many cellular processes including protein degradation. Degradation of a protein via this system involves two successive steps: ubiquitination and degradation. Ubiquitination tags the target protein with ubiquitin-like proteins (UBLs), such as ubiquitin, small ubiquitin-like modifier (SUMO) and NEDD8, via a cascade involving three enzymes: activating enzyme E1, conjugating enzyme E2 and E3 ubiquitin ligases. The proteasomes recognize the UBL-tagged substrate proteins and degrade them. Accumulating evidence indicates that allostery is a central player in the regulation of ubiquitination, as well as deubiquitination and degradation. Here, we provide an overview of the key mechanistic roles played by allostery in all steps of these processes, and highlight allosteric drugs targeting them. Throughout the review, we emphasize the crucial mechanistic role played by linkers in allosterically controlling the UPS action by biasing the sampling of the conformational space, which facilitate the catalytic reactions of the ubiquitination and degradation. Finally, we propose that allostery may similarly play key roles in the regulation of molecular machines in the cell, and as such allosteric drugs can be expected to be increasingly exploited in therapeutic regimes.


Electrophysiological effects of selective D1 and D2 dopamine receptor agonists in the medial prefrontal cortex of young and aged Fischer 344 rats.
The effects of the selective dopamine D1 and D2 receptor agonists, SKF 38393 and N-0437, respectively, on the firing rate of medial prefrontal cortex (PFC) neurons were studied in young (3-5 months old), and aged (18 and 26 months old) Fischer 344 rats. Multibarrel glass micropipettes, filled with 1 mM SKF 38393 and N-0437, were lowered into the anteromedial cortical target area (PFC) of the mesocortical dopamine system in urethane-anesthetized animals. The drug solutions were locally applied by pressure ejection. In young rats, both agonists produced dose-dependent and reversible reductions in firing rates. However, the D2 agonist was approximately 10 times more potent than the D1 agonist in suppressing firing rate. Even at the highest doses, SKF 38393 rarely produced complete cessation of firing in PFC cells. Moreover, no evidence of synergism was observed when the two drugs were simultaneously applied. PFC neurons in aged rats were significantly subsensitive to locally applied SKF 38393, whereas no change in sensitivity to N-0437 was observed. These results suggest that both D1 and D2 receptors are present in the PFC and that agonist occupancy of each of these receptors elicits an inhibition of PFC neuron discharge; furthermore, these data suggest an age-related change in D1 dopamine receptor-mediated processes with no concomitant change in processes linked to the activation of D2 dopamine processes.


Match analysis and player characteristics in rugby sevens.
Rugby sevens is a contact sport contested by two teams of seven players who compete over two 7-min halves, most frequently played in a tournament style. The IRB Sevens World Series is thought of as the preeminent rugby sevens competition in the world and has grown in competitiveness from its inception in 2000. The decision to include rugby sevens in the 2016 Olympics is likely to increase the global profile and participation in the game. Many rugby sevens players concurrently compete in 15-a-side rugby union as backs and loose forwards; however, a continued increase in the popularity of rugby sevens will likely see the emergence of the specialist rugby sevens player. Often thought of as the abbreviated version of rugby union, rugby sevens is played under nearly identical laws and on the same field dimensions as the 15-man code. However, research has shown the movement demands of rugby sevens and rugby union are dissimilar, with rugby sevens players spending a larger proportion of the game running at high intensity (≥ 5 m s(-1)). Given the dissimilarity in match demands in conjunction with differences in the competition structure between the codes, it appears the considerable depth of literature specific to performance in rugby union may be of little value for the preparation of rugby sevens players. Investigations of the physical characteristics of rugby sevens players show backs are lighter and shorter than forwards, while players across all positions possess a lean body composition. International rugby sevens players have similar speed characteristics to rugby union backs across distances of 10-30 m; however, rugby sevens players appear to have superior intermittent aerobic endurance. Despite being of likely importance, little is known of the strength and power characteristics of rugby sevens players. Research into the speed and aerobic endurance characteristics of rugby sevens players has not distinguished between backs and forwards and, as such, it is unclear whether differences exist between the position groups.


Trends in ages at key reproductive transitions in the United States, 1951-2010.
Key sexual and reproductive health milestones typically mark changing life stages with different fertility intentions and family planning needs. Knowing the typical ages at such events contributes to our understanding of changes in family formation and transition to adulthood and helps inform needs for reproductive health services. We used data from the 1982-2010 National Surveys of Family Growth and the 1995 National Survey of Adolescent Males and event history methods to examine trends over time for women and men in the median ages at several reproductive and demographic events. Women's reports indicate that age at menarche has changed little since 1951. Women's and men's median ages at first sex declined through the 1978 birth cohort, but increased slightly since then, to 17.8 years for women and 18.1 for men. The interval from first sex to first contraceptive use has narrowed, although Hispanic women have a longer interval. Age at first union (defined as the earlier of first marriage or first cohabiting relationship) has remained relatively stable, but the time between median age at first sex and median age at first birth has increased to 9.2 years for women and 11.4 for men. For some women and men born in the late 1970s, median age at first birth was earlier than median age at first marriage for the first time in at least the past several decades. The large majority of the reproductive years are spent sexually active. Thus, women have a lengthy period during which they require effective methods. In particular, the period between first sex and first childbearing has lengthened, but long-acting method use, although increasing, has not kept up with this shift. Moving the contraceptive method mix toward underutilized but highly effective contraceptive methods has the potential to reduce the unintended pregnancy rate.


Cerebellar plasticity and associative memories are controlled by perineuronal nets.
Perineuronal nets (PNNs) are assemblies of extracellular matrix molecules, which surround the cell body and dendrites of many types of neuron and regulate neural plasticity. PNNs are prominently expressed around neurons of the deep cerebellar nuclei (DCN), but their role in adult cerebellar plasticity and behavior is far from clear. Here we show that PNNs in the mouse DCN are diminished during eyeblink conditioning (EBC), a form of associative motor learning that depends on DCN plasticity. When memories are fully acquired, PNNs are restored. Enzymatic digestion of PNNs in the DCN improves EBC learning, but intact PNNs are necessary for memory retention. At the structural level, PNN removal induces significant synaptic rearrangements in vivo, resulting in increased inhibition of DCN baseline activity in awake behaving mice. Together, these results demonstrate that PNNs are critical players in the regulation of cerebellar circuitry and function.