Gout Can Increase the Risk of Receiving Rotator Cuff Tear Repair Surgery.
Gout commonly involves joint inflammation, and clinical epidemiological studies on involved tendons are scant. Rotator cuff tears are the most common cause of shoulder disability, and surgery is one of the choices often adopted to regain previous function. To investigate the risk of receiving rotator cuff repair surgery among patients with gout and to analyze possible risk factors to design an effective prevention strategy. Cohort study; Level of evidence, 3. The authors studied a 7-year longitudinal follow-up of patients from the Taiwan Longitudinal Health Insurance Database 2005 (LHID2005). This included a cohort of patients who received a diagnosis of gout during 2004-2008 (gout cohort) and a cohort matched by propensity scores (control cohort). A 2-stage approach that used the National Health Interview Survey 2005 was used to obtain missing confounding variables from the LHID2005. The crude hazard ratio (HR) and adjusted HR were estimated between the gout and control cohorts. The gout and control cohorts comprised 32,723 patients with gout and 65,446 people matched at a ratio of 1:2. The incidence of rotator cuff repair was 31 and 18 per 100,000 person-years in the gout and control cohorts, respectively. The crude HR for rotator cuff repair in the gout cohort was 1.73 (95% confidence interval [CI], 1.23-2.44; P < .01) during the 7-year follow-up period. After adjustment for covariates by use of the 2-stage approach, the propensity score calibration-adjusted HR was 1.60 (95% CI, 1.12-2.29; P < .01) in the gout cohort. Further analysis revealed that the adjusted HR was 1.73 (95% CI, 1.20-2.50; P < .001) among patients with gout who did not take hypouricemic medication and 2.70 (95% CI, 1.31-5.59; P < .01) for patients with gout aged 50 years or younger. Patients with gout, particularly those aged 50 years or younger and without hypouricemic medication control, are at a relatively higher risk of receiving rotator cuff repair surgery. Strict control of uric acid levels with hypouricemic medication may effectively reduce the risk of rotator cuff repair.


Nuclear-receptor-mediated telomere insertion leads to genome instability in ALT cancers.
The breakage-fusion-bridge cycle is a classical mechanism of telomere-driven genome instability in which dysfunctional telomeres are fused to other chromosomal extremities, creating dicentric chromosomes that eventually break at mitosis. Here, we uncover a distinct pathway of telomere-driven genome instability, specifically occurring in cells that maintain telomeres with the alternative lengthening of telomeres mechanism. We show that, in these cells, telomeric DNA is added to multiple discrete sites throughout the genome, corresponding to regions regulated by NR2C/F transcription factors. These proteins drive local telomere DNA addition by recruiting telomeric chromatin. This mechanism, which we name targeted telomere insertion (TTI), generates potential common fragile sites that destabilize the genome. We propose that TTI driven by NR2C/F proteins contributes to the formation of complex karyotypes in ALT tumors.


Cationic liposomes produced via ethanol injection method for dendritic cell therapy.
Cationic liposomes can be designed and developed in order to be an efficient gene delivery system for mammalian cells. Dendritic cell (DC) vaccines can be used to treat cancer, as cationic liposomes can deliver tumor antigens to cells while cells remain active. However, most methods used for liposome production are not able to reproduce in large scale the physicochemical and biological properties of liposomes produced in laboratory scale. In this context, ethanol injection method achieved promising results, although requiring post-treatment for size reduction and/or to remove residual ethanol. Thus, the purpose of this study was to generate cationic liposomes suitable for gene therapies via ethanol injection method in only one step (VEI) and compared to those submitted to a size reduction processes by microfluidization (MFV). For this, the method to produce cationic liposomes composed of egg phosphatidylcholine (EPC), 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and 1,2-dioleoylphosphatidylethanolamine (DOPE) was optimized using a statistical design approach. As a result, the size of VEI decreased from 290 nm to 110 nm and the polydispersity from 0.54 to 0.17. In the case of MFV, size decreased from 128 nm to 107 nm and polydispersity from 0.40 to 0.18. ST and MFV before and after optimization were also characterized in terms of morphology by transmission electron microscopy (TEM) and structure by differential scanning calorimetry (DSC). Finally, to show their potential in gene/immune therapies applications, DCs were stimulated by such liposomes. Cells internalized liposomes, increasing expression of the costimulatory molecule CD86 and inducing T lymphocyte proliferation.


Niche partitioning in three sympatric congeneric species of dragonfly, Orthetrum chrysostigma, O. coerulescens anceps, and O. nitidinerve: the importance of microhabitat.
Habitat heterogeneity has been shown to promote co-existence of closely related species. Based on this concept, a field study was conducted on the niche partitioning of three territorial congeneric species of skimmers (Anisoptera: Libellulidae) in Northeast Algeria during the breeding season of 2011. According to their size, there is a descending hierarchy between Orthetrum nitidinerve Sélys, O. chrysostigma (Burmeister), and O. coerulescens anceps (Schneider). After being marked and surveyed, the two latter species had the same breeding behavior sequence. Knowing that they had almost the same size, such species could not co-occur in the same habitat according to the competitive exclusion principle. The spatial distribution of the three species was investigated at two different microhabitats, and it was found that these two species were actually isolated at this scale. O. chrysostigma and O. nitidinerve preferred open areas, while O. c. anceps occurred in highly vegetated waters. This study highlights the role of microhabitat in community structure as an important niche axis that maintains closely related species in the same habitat.


Controllability of multiplex, multi-time-scale networks.
The paradigm of layered networks is used to describe many real-world systems, from biological networks to social organizations and transportation systems. While recently there has been much progress in understanding the general properties of multilayer networks, our understanding of how to control such systems remains limited. One fundamental aspect that makes this endeavor challenging is that each layer can operate at a different time scale; thus, we cannot directly apply standard ideas from structural control theory of individual networks. Here we address the problem of controlling multilayer and multi-time-scale networks focusing on two-layer multiplex networks with one-to-one interlayer coupling. We investigate the practically relevant case when the control signal is applied to the nodes of one layer. We develop a theory based on disjoint path covers to determine the minimum number of inputs (N_{i}) necessary for full control. We show that if both layers operate on the same time scale, then the network structure of both layers equally affect controllability. In the presence of time-scale separation, controllability is enhanced if the controller interacts with the faster layer: N_{i} decreases as the time-scale difference increases up to a critical time-scale difference, above which N_{i} remains constant and is completely determined by the faster layer. We show that the critical time-scale difference is large if layer I is easy and layer II is hard to control in isolation. In contrast, control becomes increasingly difficult if the controller interacts with the layer operating on the slower time scale and increasing time-scale separation leads to increased N_{i}, again up to a critical value, above which N_{i} still depends on the structure of both layers. This critical value is largely determined by the longest path in the faster layer that does not involve cycles. By identifying the underlying mechanisms that connect time-scale difference and controllability for a simplified model, we provide crucial insight into disentangling how our ability to control real interacting complex systems is affected by a variety of sources of complexity.


Experiences of neurological surgeons with malpractice lawsuits.
As a specialty that treats acute pathology and refractory pain, neurosurgery is at risk for high liability, making the practice of defensive medicine quite common. The extent to which the practice of defensive medicine is linked to experience with malpractice lawsuits remains unclear. The aims of this study were to clarify this by surveying neurosurgeons about the frequency of experiencing medical lawsuits and to show how neurosurgeons reflect on facing such lawsuits. A survey consisting of 24 questions was distributed among members of the Congress of Neurological Surgeons. The survey consisted of four parts: 1) demographics of participants; 2) the way malpractice lawsuits affect the way respondents practice medicine; 3) experiences with medical malpractice lawsuits; and 4) the effect of the medical malpractice environment on one's own practice of medicine. There were a total of 490 survey respondents, 83.5% of whom were employed in the US. Of the respondents, 39.5% stated they were frequently or always concerned about being sued, and 77.4% stated their fear had led to a change in how they practice medicine. For 58.4%, this change led to the practice of defensive medicine, while for others it led to more extensive documentation (14.3%) and/or to referring or dropping complex cases (12.4%).Among the respondents, 80.9% at some time were named in a medical malpractice lawsuit and 12.3% more than 10 times. The main concerns expressed about being sued included losing confidence and practicing defensive medicine (17.8%), personal assets being at risk (16.9%), and being named in the National Practitioner Data Bank (15.6%). Given the medical malpractice environment, 58.7% of respondents considered referring complex patient cases, whereas 36.5% considered leaving the practice of medicine. The fear of being sued (OR 4.06, 95% CI 2.53-6.51) and the consideration of limiting the scope of practice (OR 3.08, 1.80-5.20) were both independently associated with higher odds of considering leaving the practice of medicine. The current medicolegal landscape has a profound impact on neurosurgical practice. The fear of being sued, the financial aspects of practicing defensive medicine, and the proportion of neurosurgeons who are considering leaving the practice of medicine emphasize the need for a shift in the medicolegal landscape to a system in which fear of being sued does not play a dominant role and the interests of patients are protected.


Life Cycle of the Trout Cecal Nematode, Truttaedacnitis truttae (Nematoda: Cucullanidae): Experimental and Field Observations.
Truttaedacnitis truttae is a cucullanid nematode of primarily salmonine fishes. Brown trout (Salmo trutta) in Europe reportedly become parasitized by ingesting lampreys (Lampetra planeri) carrying infective larvae. However, our field and laboratory observations suggested that North American specimens of T. truttae have an alternative life cycle. High abundances and potential impact of T. truttae in rainbow trout, Oncorhynchus mykiss, in the Colorado River drainage in Grand Canyon, where there are no lampreys, prompted a study on the transmission dynamics of this nematode. Eggs of T. truttae, collected from live gravid females, were incubated in the laboratory. Snails, Physa gyrina and Lymnaea sp., were exposed to T. truttae larvae 3-4 wk later. Active larvae of T. truttae were observed penetrating the intestinal wall of exposed snails, and worm larvae were found in the visceral tissues when examined 1 wk after exposure. Larvae in snails showed little growth and development 2 wk later and corresponded to L3 larvae. Infected snails were fed to hatchery-reared juvenile rainbow trout. Developing stages were subsequently found in the mucosal lining and lumen of trout intestines. Adult male and female (gravid) worms were found in the ceca of trout examined 5-6 mo after consuming infected snails. Larvae found in pepsin/trypsin digests and mucosal scrapings from wild, naturally infected, trout corroborate laboratory findings. Screening of Physa sp. and gammarids collected from Colorado River, Grand Canyon, for natural infections with T. truttae using the ITS1 rDNA marker gave positive results. Truttaedacnitis truttae is the second species, after Truttaedacnitis clitellarius of lake sturgeon, capable of using a snail first intermediate/paratenic host and is similar to several other cucullanids in having a histotropic phase of development in the definitive fish host.


The role of informal caregivers at the end of life: providing support through Advance Care Planning.
In the palliative phase of a person's illness, the needs of the patient and informal caregivers are paramount. Caregivers often look to palliative care practitioners for support, guidance and emotional comfort. This article reviews the role of the informal caregiver at the end of life, summarising their concerns and needs. The paper also highlights caregiver difficulties in making the coping transition when the patient's medical condition becomes advanced and they require end-of-life care. The focus of the review is placed on discussions about Advanced Care Planning (ACP). ACP is a complex legal, ethical and practical issue that can enable caregivers to consider future strategies and enable them to provide high quality care at the end of life. The paper looks at some of the ethical and legal issues associated with this sensitive end-of-life issue.


Rapamycin nanoparticles localize in diseased lung vasculature and prevent pulmonary arterial hypertension.
Vascular remodeling resulting from pulmonary arterial hypertension (PAH) leads to endothelial fenestrations. This feature can be exploited by nanoparticles (NP), allowing them to extravasate from circulation and accumulate in remodeled pulmonary vessels. Hyperactivation of the mTOR pathway in PAH drives pulmonary arterial smooth muscle cell proliferation. We hypothesized that rapamycin (RAP)-loaded NPs, an mTOR inhibitor, would accumulate in diseased lungs, selectively targeting vascular mTOR and preventing PAH progression. RAP poly(ethylene glycol)-block-poly(ε-caprolactone) (PEG-PCL) NPs were fabricated. NP accumulation and efficacy were examined in a rat monocrotaline model of PAH. Following intravenous (IV) administration, NP accumulation in diseased lungs was verified via LC/MS analysis and confocal imaging. Pulmonary arteriole thickness, right ventricular systolic pressures, and ventricular remodeling were determined to assess the therapeutic potential of RAP NPs. Monocrotaline-exposed rats showed increased NP accumulation within lungs compared to healthy controls, with NPs present to a high extent within pulmonary perivascular regions. RAP, in both free and NP form, attenuated PAH development, with histological analysis revealing minimal changes in pulmonary arteriole thickness and no ventricular remodeling. Importantly, NP-treated rats showed reduced systemic side effects compared to free RAP. This study demonstrates the potential for nanoparticles to significantly impact PAH through site-specific delivery of therapeutics.


Depressive symptoms and headaches in relation to proximity of residence to an alternating-current transmission line right-of-way.
Electric power transmission lines have become objects of public controversy. Hypotheses have linked neurobehavioral effects to the electric and magnetic fields that these lines produce. The authors conducted a telephone interview survey in November 1987 to assess the prevalence of depressive symptoms and headache in relation to proximity of residence to an alternating-current transmission line in the United States. Proximity to the line, defined as residing on a property abutting the right-of-way or being able to see the towers from one's house or yard, was positively associated with a measure of depressive symptoms. The association was not explained by demographic variables associated with depression or by attitudes about power lines or other environmental issues. The estimated prevalence odds ratio was 2.8 (95% confidence interval (CI) 1.6-5.1). The estimate did not change appreciably when the definitions of depressive symptoms or of proximity to the line were altered. Nonmigraine headaches had a weaker association with proximity to the line (odds ratio = 1.5, 95% CI 0.76-2.8), and self-reported migraine headaches exhibited no association (odds ratio = 0.99, 95% CI 0.29-3.4). Additional studies of psychological and behavioral measures should be conducted in relation to electric and magnetic fields, with a strong emphasis on improved exposure assessment.


Cardiac micro-computed tomography for morphological and functional phenotyping of muscle LIM protein null mice.
The purpose of this study was to investigate the use of micro-computed tomography (micro-CT) for morphological and functional phenotyping of muscle LIM protein (MLP) null mice and to compare micro-CT with M-mode echocardiography. MLP null mice and controls were imaged using both micro-CT and M-mode echocardiography. For micro-CT, we used a custom-built scanner. Following a single intravenous injection of a blood pool contrast agent (Fenestra VC, ART Advanced Research Technologies, Saint-Laurent, QC) and using a cardiorespiratory gating, we acquired eight phases of the cardiac cycle (every 15 ms) and reconstructed three-dimensional data sets with 94-micron isotropic resolution. Wall thickness and volumetric measurements of the left ventricle were performed, and cardiac function was estimated. Micro-CT and M-mode echocardiography showed both morphological and functional aspects that separate MLP null mice from controls. End-diastolic and -systolic volumes were increased significantly three- and fivefold, respectively, in the MLP null mice versus controls. Ejection fraction was reduced by an average of 32% in MLP null mice. The data analysis shows that two imaging modalities provided different results partly owing to the difference in anesthesia regimens. Other sources of errors for micro-CT are also analyzed. Micro-CT can provide the four-dimensional data (three-dimensional isotropic volumes over time) required for morphological and functional phenotyping in mice.


A systematic review of the clinical effectiveness and cost-effectiveness of sensory, psychological and behavioural interventions for managing agitation in older adults with dementia.
Agitation is common, persistent and distressing in dementia and is linked with care breakdown. Psychotropic medication is often ineffective or harmful, but the evidence regarding non-pharmacological interventions is unclear. We systematically reviewed and synthesised the evidence for clinical effectiveness and cost-effectiveness of non-pharmacological interventions for reducing agitation in dementia, considering dementia severity, the setting, the person with whom the intervention is implemented, whether the effects are immediate or longer term, and cost-effectiveness. We searched twice using relevant search terms (9 August 2011 and 12 June 2012) in Web of Knowledge (incorporating MEDLINE); EMBASE; British Nursing Index; the Health Technology Assessment programme database; PsycINFO; NHS Evidence; System for Information on Grey Literature; The Stationery Office Official Documents website; The Stationery National Technical Information Service; Cumulative Index to Nursing and Allied Health Literature; and The Cochrane Library. We also searched Cochrane reviews of interventions for behaviour in dementia, included papers' references, and contacted authors about 'missed' studies. We included quantitative studies, evaluating non-pharmacological interventions for agitation in dementia, in all settings. We rated quality, prioritising higher-quality studies. We separated results by intervention type and agitation level. As we were unable to meta-analyse results except for light therapy, we present a qualitative evidence synthesis. In addition, we calculated standardised effect sizes (SESs) with available data, to compare heterogeneous interventions. In the health economic analysis, we reviewed economic studies, calculated the cost of effective interventions from the effectiveness review, calculated the incremental cost per unit improvement in agitation, used data from a cohort study to evaluate the relationship between health and social care costs and health-related quality of life (DEMQOL-Proxy-U scores) and developed a new cost-effectiveness model. We included 160 out of 1916 papers screened. Supervised person-centred care, communication skills (SES = -1.8 to -0.3) or modified dementia care mapping (DCM) with implementing plans (SES = -1.4 to -0.6) were all efficacious at reducing clinically significant agitation in care home residents, both immediately and up to 6 months afterwards. In care home residents, during interventions but not at follow-up, activities (SES = -0.8 to -0.6) and music therapy (SES = -0.8 to -0.5) by protocol reduced mean levels of agitation; sensory intervention (SES = -1.3 to -0.6) reduced mean and clinically significant symptoms. Advantages were not demonstrated with 'therapeutic touch' or individualised activity. Aromatherapy and light therapy did not show clinical effectiveness. Training family carers in behavioural or cognitive interventions did not decrease severe agitation. The few studies reporting activities of daily living or quality-of-life outcomes found no improvement, even when agitation had improved. We identified two health economic studies. Costs of interventions which significantly impacted on agitation were activities, £80-696; music therapy, £13-27; sensory interventions, £3-527; and training paid caregivers in person-centred care or communication skills with or without behavioural management training and DCM, £31-339. Among the 11 interventions that were evaluated using the Cohen-Mansfield Agitation Inventory (CMAI), the incremental cost per unit reduction in CMAI score ranged from £162 to £3480 for activities, £4 for music therapy, £24 to £143 for sensory interventions, and £6 to £62 for training paid caregivers in person-centred care or communication skills with or without behavioural management training and DCM. Health and social care costs ranged from around £7000 over 3 months in people without clinically significant agitation symptoms to around £15,000 at the most severe agitation levels. There is some evidence that DEMQOL-Proxy-U scores decline with Neuropsychiatric Inventory agitation scores. A multicomponent intervention in participants with mild to moderate dementia had a positive monetary net benefit and a 82.2% probability of being cost-effective at a maximum willingness to pay for a quality-adjusted life-year of £20,000 and a 83.18% probability at a value of £30,000. Although there were some high-quality studies, there were only 33 reasonably sized (> 45 participants) randomised controlled trials, and lack of evidence means that we cannot comment on many interventions' effectiveness. There were no hospital studies and few studies in people's homes. More health economic data are needed. Person-centred care, communication skills and DCM (all with supervision), sensory therapy activities, and structured music therapies reduce agitation in care-home dementia residents. Future interventions should change care home culture through staff training and permanently implement evidence-based treatments and evaluate health economics. There is a need for further work on interventions for agitation in people with dementia living in their own homes. The study was registered as PROSPERO no. CRD42011001370. The National Institute for Health Research Health Technology Assessment programme.


Selective serotonin reuptake inhibitors ameliorate MEGF10 myopathy.
MEGF10 myopathy is a rare inherited muscle disease that is named after the causative gene, MEGF10. The classic phenotype, early onset myopathy, areflexia, respiratory distress and dysphagia, is severe and immediately life-threatening. There are no disease-modifying therapies. We performed a small molecule screen and follow-up studies to seek a novel therapy. A primary in vitro drug screen assessed cellular proliferation patterns in Megf10-deficient myoblasts. Secondary evaluations were performed on primary screen hits using myoblasts derived from Megf10-/- mice, induced pluripotent stem cell-derived myoblasts from MEGF10 myopathy patients, mutant Drosophila that are deficient in the homologue of MEGF10 (Drpr) and megf10 mutant zebrafish. The screen yielded two promising candidates that are both selective serotonin reuptake inhibitors (SSRIs), sertraline and escitalopram. In depth follow-up analyses demonstrated that sertraline was highly effective in alleviating abnormalities across multiple models of the disease including mouse myoblast, human myoblast, Drosophila and zebrafish models. Sertraline also restored deficiencies of Notch1 in disease models. We conclude that SSRIs show promise as potential therapeutic compounds for MEGF10 myopathy, especially sertraline. The mechanism of action may involve the Notch pathway.


Epigenetics and immunotherapy: The current state of play.
Cancer cells employ a number of mechanisms to escape immunosurveillance and facilitate tumour progression. The recent explosion of interest in immunotherapy, especially immune checkpoint blockade, is a result of discoveries about the fundamental ligand-receptor interactions that occur between immune and cancer cells within the tumour microenvironment. Distinct ligands expressed by cancer cells engage with cell surface receptors on immune cells, triggering inhibitory pathways (such as PD-1/PD-L1) that render immune cells immunologically tolerant. Importantly, recent studies on the role of epigenetics in immune evasion have exposed a key role for epigenetic modulators in augmenting the tumour microenvironment and restoring immune recognition and immunogenicity. Epigenetic drugs such as DNA methyltransferase and histone deacetylase inhibitors can reverse immune suppression via several mechanisms such as enhancing expression of tumour-associated antigens, components of the antigen processing and presenting machinery pathways, immune checkpoint inhibitors, chemokines, and other immune-related genes. These discoveries have established a highly promising basis for studies using combined epigenetic and immunotherapeutic agents as anti-cancer therapies. In this review, we discuss the exciting role of epigenetic immunomodulation in tumour immune escape, emphasising its significance in priming and sensitising the host immune system to immunotherapies through mechanisms such as the activation of the viral defence pathway. With this background in mind, we highlight the promise of combined epigenetic therapy and immunotherapy, focusing on immune checkpoint blockade, to improve outcomes for patients with many different cancer types.


Metagenomic and metatranscriptomic inventories of the lower Amazon River, May 2011.
The Amazon River runs nearly 6500 km across the South American continent before emptying into the western tropical North Atlantic Ocean. In terms of both volume and watershed area, it is the world's largest riverine system, affecting elemental cycling on a global scale. A quantitative inventory of genes and transcripts benchmarked with internal standards was obtained at five stations in the lower Amazon River during May 2011. At each station, metagenomes and metatranscriptomes were obtained in duplicate for two microbial size fractions (free-living, 0.2 to 2.0 μm; particle-associated, 2.0 to 297 μm) using 150 × 150 paired-end Illumina sequencing. Forty eight sample datasets were obtained, averaging 15 × 10(6) potential protein-encoding reads each (730 × 10(6) total). Prokaryotic metagenomes and metatranscriptomes were dominated by members of the phyla Actinobacteria, Planctomycetes, Betaproteobacteria, Verrucomicrobia, Nitrospirae, and Acidobacteria. The actinobacterium SCGC AAA027-L06 reference genome recruited the greatest number of reads overall, with this single bin contributing an average of 50 billion genes and 500 million transcripts per liter of river water. Several dominant taxa were unevenly distributed between the free-living and particle-associated size fractions, such as a particle-associated bias for reads binning to planctomycete Schlesneria paludicola and a free-living bias for actinobacterium SCGC AAA027-L06. Gene expression ratios (transcripts to gene copy ratio) increased downstream from Óbidos to Macapá and Belém, indicating higher per cell activity of Amazon River bacteria and archaea as river water approached the ocean. This inventory of riverine microbial genes and transcripts, benchmarked with internal standards for full quantitation, provides an unparalleled window into microbial taxa and functions in the globally important Amazon River ecosystem.


Marine natural products.
This review covers the literature published in 2013 for marine natural products (MNPs), with 982 citations (644 for the period January to December 2013) referring to compounds isolated from marine microorganisms and phytoplankton, green, brown and red algae, sponges, cnidarians, bryozoans, molluscs, tunicates, echinoderms, mangroves and other intertidal plants and microorganisms. The emphasis is on new compounds (1163 for 2013), together with the relevant biological activities, source organisms and country of origin. Reviews, biosynthetic studies, first syntheses, and syntheses that lead to the revision of structures or stereochemistries, have been included.


Endogenous prostaglandin synthesis inhibitor in the renal cortex. Effects on production of prostacyclin by renal blood vessels.
1. The capacity of various tissues of the porcine kidney to convert [1-14C]arachidonic acid into radiolabelled prostaglandins was studied. 2. Only after removal from the cortical matrix, were renal blood vessels able to convert arachidonic acid into prostaglandins (primarily prostacyclin). In contrast, convoluted tubules showed a low capacity to metabolize arachidonic acid. 3. The failure to demonstrate prostaglandin synthesis by renal cortical slices is related to the presence of an inhibitor of cyclo-oxygenase. Thus the addition of renal cortical incubate to isolated vascular tissues and ram seminal vesicles inhibited their ability to synthesize prostaglandins. 4. Slices of renal medulla metabolized arachidonic acid primarily to prostaglandin F2alpha; lesser amounts of prostaglandin E2 and prostacyclin were generated. 5. The large capacity of the renal vasculature to generate prostacyclin is consistent with an important role for this prostaglandin in regulation of renin release and renal haemodynamics.


Physical-based statistical shape modeling of the levator ani.
The levator ani is the main muscular support of the pelvic floor organs and damage caused by childbirth can affect its function. The full functionality of this muscle group is still unknown but it is essential for effective surgical planning. To elucidate its functional significance, a physical-based statistical shape model was built from the levator ani surfaces of 15 subjects scanned in an open access scanner. Simulation of dynamic exercises was performed on the resulting surfaces with finite element analysis. Statistical shape modeling was performed on the training set consisting of the original and simulated shapes along with thickness and strain distributions. Simulation results are presented on 15 subjects. The statistical shape model shows good correspondence to inter- and intra-subject shape variability, with the modes of variation highlighting movement in the posterior of the levator ani as well as in the levator arms. Strain distribution plots and the modes of variation show results that correspond to clinical findings. Further validation of the technique and a repeatability test were performed on four subjects with internal global pressure readings taken from a perineometer and five patients suffering from minor pelvic floor disorders due to obstructed defaecation. A Mann-Whitney nonparametric test was used to compare the normal model fitting to the two subject groups.


Characterization of small ruminant lentivirus A4 subtype isolates and assessment of their pathogenic potential in naturally infected goats.
Small ruminant lentiviruses escaping efficient serological detection are still circulating in Swiss goats in spite of a long eradication campaign that essentially eliminated clinical cases of caprine arthritis encephalitis in the country. This strongly suggests that the circulating viruses are avirulent for goats.To test this hypothesis, we isolated circulating viruses from naturally infected animals and tested the in vitro and in vivo characteristics of these field isolates. Viruses were isolated from primary macrophage cultures. The presence of lentiviruses in the culture supernatants was monitored by reverse transcriptase assay. Isolates were passaged in different cells and their cytopathogenic effects monitored by microscopy. Proviral load was quantified by real-time PCR using customized primer and probes. Statistical analysis comprised Analysis of Variance and Bonferroni Multiple Comparison Test. The isolated viruses belonged to the small ruminant lentiviruses A4 subtype that appears to be prominent in Switzerland. The 4 isolates replicated very efficiently in macrophages, displaying heterogeneous phenotypes, with two isolates showing a pronounced cytopathogenicity for these cells. By contrast, all 4 isolates had a poor replication capacity in goat and sheep fibroblasts. The proviral loads in the peripheral blood and, in particular, in the mammary gland were surprisingly high compared to previous observations. Nevertheless, these viruses appear to be of low virulence for goats except for the mammary gland were histopathological changes were observed. Small ruminant lentiviruses continue to circulate in Switzerland despite a long and expensive caprine arthritis encephalitis virus eradication campaign. We isolated 4 of these lentiviruses and confirmed their phylogenetic association with the prominent A4 subtype. The pathological and histopathological analysis of the infected animals supported the hypothesis that these A4 viruses are of low pathogenicity for goats, with, however, a caveat about the potentially detrimental effects on the mammary gland. Moreover, the high proviral load detected indicates that the immune system of the animals cannot control the infection and this, combined with the phenotypic plasticity observed in vitro, strongly argues in favour of a continuous and precise monitoring of these SRLV to avoid the risk of jeopardizing a long eradication campaign.


Isolation and properties of liver cell nucleoli.
1. The significance of the term nucleolus has been discussed. 2. A detailed method for the isolation of nucleoli from already isolated rat or cat liver nuclei has been presented. 3. The presence of DNA in isolated liver cell nucleoli has been indicated by histochemical methods. 4. The percentages of DNA and RNA in the isolated nucleoli have been determined by chemical analysis. 5. The specific activities of aldolase, arginase, and catalase have been determined for two subnuclear fractions and for the isolated nucleoli of rat and cat liver, and the relative amounts of these enzymes in the same subnuclear fractions and nucleoli of rat liver have been measured. 6. The significance of the above findings has been discussed and consideration has been given to what types of isolated nuclei might best serve as starting material for the isolation of nucleoli. 7. A new hypothesis has been presented that nucleoli of the liver cell type may function primarily in furnishing (directly or indirectly) templates for the synthesis of the particular enzymes that must govern the chemistry of mitosis.


Pancreatic stent prophylaxis of post endoscopic retrograde cholangiopancreatography pancreatitis: spontaneous migration rates and clinical outcomes.
Pancreatic duct (PD) stents diminish the risk of post endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP) in high-risk patients; 3 Fr stents are reported to spontaneously migrate at a significantly higher rate than 5 Fr stents in a cohort of mostly sphincter of Oddi (SOD) patients. We sought to assess spontaneous migration rates of 5 Fr and 7 Fr stents and effectiveness in preventing PEP in a diverse group of high risk patients. A total of 4,332 ERCP exams performed between January 2002 and August 2005 were reviewed to identify patients undergoing PD stent placement. Follow-up was obtained from electronic medical records and contact with referring MDs. Plain abdominal radiographs were used to document stent passage. PD stents for PEP prophylaxis were placed in 246 exams (232 patients) undergoing: PD (major or minor) sphincterotomy (84), ampullectomy (50), SOD (46), bile duct precut (35), papillary stenosis balloon dilation (9) and difficult cannulation (8). Stents placed: 218 5-Fr (140 were 3 cm long and 78 =or> 5 cm long) and 28 7-Fr (12 were 3 cm long, 16=or> 5cm long). Follow-up was available in 197 (171 5-Fr, 26 7-Fr) of 246 placements (80%). Twenty of 171 5-Fr stents were electively removed via EGD within=or< 24 h per endoscopist preference and were not included in analysis; 128 of the remaining 151 5-Fr stents (85%) spontaneously migrated by (or within) median of 8 days and 23 failed to pass and required EGD removal. Of 26 7-Fr stents one was electively removed =or< 24 h later; of the remaining 25, 15 (60%) spontaneously migrated by median of 16 days, 10 required EGD removal. The spontaneous migration rate of 5 Fr stents was: 1) significantly higher than 7 Fr stents; 2) significantly higher than the previously reported 67% passage rate of 5 Fr stents; and 3) similar to the previously reported 86% passage rate of 3 Fr stents. PEP occurred in 15% (n=36: 24 mild, 11 moderate, 1 severe). The spontaneous dislodgement rate of 5 Fr stents in patients where the indication is primarily non-SOD is approximately 85% - significantly higher than previously reported and similar to the reported rate of spontaneous dislodgement of 3 Fr stents in SOD patients; 5 Fr stents migrate spontaneously earlier and more frequently than 7 Fr stents.


Diabetes service provision: a qualitative study of the experiences and views of Pakistani and Indian patients with Type 2 diabetes.
To explore Pakistani and Indian patients' experiences of, and views about, diabetes services in order to inform the development of culturally sensitive services. Qualitative, interview study involving 23 Pakistani and nine Indian patients with Type 2 diabetes recruited from general practices and the local community in Edinburgh, Scotland. Data collection and analysis occurred concurrently and recruitment continued until no new themes emerged from the interviews. Respondents expressed gratitude for the availability of free diabetes services in Britain, as they were used to having to pay to access health care on the Indian subcontinent. Most looked to services for the prompt detection and treatment of complications, rather than the provision of advice about managing their condition. As respondents attached importance to receiving physical examinations, they could be disappointed when these were not offered by health-care professionals. They disliked relying on interpreters and identified a need for bilingual professionals with whom they could discuss their diabetes care directly. Gratitude for free services in Britain may instil a sense of indebtedness which makes it difficult for Pakistanis and Indians to be critical of their diabetes care. Health-care professionals may need to describe their roles carefully, and explain how different diabetes services fit together, to avoid Pakistani and Indian patients perceiving treatment as unsatisfactory. Whilst linkworker schemes may meet patients' need to receive culturally sensitive information in their first language, work is needed to assess their effectiveness and sustainability.


Myosin-induced acute myocarditis is a T cell-mediated disease.
The heart is a target organ in several autoimmune diseases, and therefore it is important to understand more about the effector cells involved in immune-mediated mechanisms of myocardial cell death. Because immune T lymphocytes are central to many immune responses, we wanted to study the role of T cells in causing cardiac specific inflammation. We used purified mouse cardiac myosin to cause acute myocarditis in mice. The adoptive transfer of purified T cells from C.B-17 mice with active myocarditis to SCID recipients successfully transferred the disease into SCID hosts. In contrast, transfer of serum with high-titer antimyosin antibodies to SCID hosts did not cause myocarditis. Using mAb to deplete A/J mice of CD4+ T cells, we showed that these mice were protected against the induction of myocarditis. Depletion of CD8+ T cells reduced the severity of inflammation but did not prevent induction of myocarditis. We were also able to prevent the induction of myocarditis using major histocompatibility class II protein-binding, nonimmunogenic, competitor peptides. These blocking studies also indicated that in H-2k mice, myocarditis is an I-Ak-restricted disease, and provided further evidence that CD4+ T cells are critical to the induction of disease. Together, these studies provide direct evidence that myosin-induced myocarditis is a T cell-mediated disease.


A Microbiological Map of the Healthy Equine Gastrointestinal Tract.
Horses are exquisitely sensitive to non-specific gastrointestinal disturbances as well as systemic and extraintestinal conditions related to gut health, yet minimal data are available regarding the composition of the microbiota present in the equine stomach, small intestine, and cecum and their relation to fecal microbiota. Moreover, there is minimal information regarding the concordance of the luminal and mucosal microbial communities throughout the equine gut. Illumina-based 16S rRNA gene amplicon sequencing of the luminal and mucosal microbiota present in seven regions of the gastrointestinal tract of nine healthy adult horses revealed a distinct compositional divide between the small and large intestines. This disparity in composition was more pronounced within the luminal contents, but was also detected within mucosal populations. Moreover, the uniformity of the gut microbiota was much higher in the cecum and colon relative to that in the stomach, jejunum and ileum, despite a significantly higher number of unique sequences detected in the colon. Collectively, the current data suggest that while colonic samples (a proxy for feces) may provide a reasonable profile of the luminal contents of the healthy equine large intestine, they are not informative with regard to the contents of the stomach or small intestine. In contrast to the distinct difference between the highly variable upper gastrointestinal tract microbiota and relatively uniform large bowel microbiota present within the lumen, these data also demonstrate a regional continuity present in mucosal microbial communities throughout the length of the equine gut.


Recruitment maneuvers for acute lung injury: a systematic review.
There are conflicting data regarding the safety and efficacy of recruitment maneuvers (RMs) in patients with acute lung injury (ALI). To summarize the physiologic effects and adverse events in adult patients with ALI receiving RMs. Systematic review of case series, observational studies, and randomized clinical trials with pooling of study-level data. Forty studies (1,185 patients) met inclusion criteria. Oxygenation (31 studies; 636 patients) was significantly increased after an RM (PaO2): 106 versus 193 mm Hg, P = 0.001; and PaO2/FiO2 ratio: 139 versus 251 mm Hg, P < 0.001). There were no persistent, clinically significant changes in hemodynamic parameters after an RM. Ventilatory parameters (32 studies; 548 patients) were not significantly altered by an RM, except for higher PEEP post-RM (11 versus 16 cm H2O; P = 0.02). Hypotension (12%) and desaturation (9%) were the most common adverse events (31 studies; 985 patients). Serious adverse events (e.g., barotrauma [1%] and arrhythmias [1%]) were infrequent. Only 10 (1%) patients had their RMs terminated prematurely due to adverse events. Adult patients with ALI receiving RMs experienced a significant increase in oxygenation, with few serious adverse events. Transient hypotension and desaturation during RMs is common but is self-limited without serious short-term sequelae. Given the uncertain benefit of transient oxygenation improvements in patients with ALI and the lack of information on their influence on clinical outcomes, the routine use of RMs cannot be recommended or discouraged at this time. RMs should be considered for use on an individualized basis in patients with ALI who have life-threatening hypoxemia.


Systematic review of ablative neurosurgical techniques for the treatment of trigeminal neuralgia.
There are no randomized controlled trials comparing retrogasserian percutaneous radiofrequency thermocoagulation, glycerol rhizolysis, balloon compression of the gasserian ganglion, and stereotactic radiosurgery, nor are there systematic reviews using predefined quality criteria. The objective of this study was to systematically identify all of the studies reporting outcomes and complications of ablative techniques for treatment of trigeminal neuralgia, from the development of electronic databases, and to evaluate them with predefined quality criteria. Inclusion criteria for the outcome analysis included thorough demographic documentation, defined diagnostic and outcome criteria, a minimum of 30 patients treated and median/mean follow-up times of 12 months, not more than 20% of patients lost to follow-up monitoring, Kaplan-Meier actuarial analysis of individual procedures, less than 10% of patients retreated because of failure or early recurrence, and a minimal dose of 70 Gy for stereotactic radiosurgery. High-quality studies with no actuarial analysis were used for the evaluation of complications. Of 175 studies identified, 9 could be used to evaluate rates of complete pain relief on a yearly basis and 22 could be used to evaluate complications. In mixed series, radiofrequency thermocoagulation offered higher rates of complete pain relief, compared with glycerol rhizolysis and stereotactic radiosurgery, although it demonstrated the greatest number of complications. Radiofrequency thermocoagulation offers the highest rates of complete pain relief, although further data on balloon microcompression are required. It is essential that uniform outcome measures and actuarial methods be universally adopted for the reporting of surgical results. Randomized controlled trials are required to reliably evaluate new surgical techniques.


MEDIATE - Molecular DockIng at homE: Turning collaborative simulations into therapeutic solutions.
Collaborative computing has attracted great interest in the possibility of joining the efforts of researchers worldwide. Its relevance has further increased during the pandemic crisis since it allows for the strengthening of scientific collaborations while avoiding physical interactions. Thus, the E4C consortium presents the MEDIATE initiative which invited researchers to contribute via their virtual screening simulations that will be combined with AI-based consensus approaches to provide robust and method-independent predictions. The best compounds will be tested, and the biological results will be shared with the scientific community. In this paper, the MEDIATE initiative is described. This shares compounds' libraries and protein structures prepared to perform standardized virtual screenings. Preliminary analyses are also reported which provide encouraging results emphasizing the MEDIATE initiative's capacity to identify active compounds. Structure-based virtual screening is well-suited for collaborative projects provided that the participating researchers work on the same input file. Until now, such a strategy was rarely pursued and most initiatives in the field were organized as challenges. The MEDIATE platform is focused on SARS-CoV-2 targets but can be seen as a prototype which can be utilized to perform collaborative virtual screening campaigns in any therapeutic field by sharing the appropriate input files.


An independent audit of the Australian food industry's voluntary front-of-pack nutrition labelling scheme for energy-dense nutrition-poor foods.
Since 2006, the Australian food industry has promoted its front-of-pack (FOP) food labelling system-the Daily Intake Guide (DIG)-as a success story of industry self-regulation. With over 4000 products already voluntary featuring the DIG, the industry argues that government regulation of FOP nutrition labelling is simply unnecessary. However, no independent audit of the industry's self-regulation has ever been undertaken and we present the first such Australian data. Energy-dense nutrient-poor (EDNP) snacks were audited at nine Australian supermarkets, including biscuits, candy, ice creams, chocolates, crisps, sports drinks, energy drinks, flavoured milks, sweetened juices and soft drinks. In these categories nutrition labels were recorded for 728 EDNP products in various packaging sizes. The DIG was displayed on 66% of audited EDNP products but most of these (75%) did not report saturated fat and sugar content. Only generic supermarket EDNP products were likely to display saturated fat and sugar content, compared with very few branded products (48% vs 4%, P<0.001). Branded products not displaying fat and sugar content contained on average 10-times more saturated fat than those displaying such (10% vs 1% DI, P<0.001) and nearly twice as much sugar (21 vs 13% DI, P<0.05). Most Australian manufacturers of EDNP products have adopted the DIG; consistent with industry claims of widespread adoption, but almost all still avoid displaying the high saturated fat and sugar content of their products by opting for the 'energy alone' option, violating the industry's own voluntarily guidelines and highlighting serious weaknesses with the industry's self-regulation.


Cell cycle expression of two replicative DNA polymerases alpha and delta from Schizosaccharomyces pombe.
We have investigated the expression of two Schizosaccharomyces pombe replicative DNA polymerases alpha and delta during the cell cycle. The pol alpha+ and pol delta+ genes encoding DNA polymerases alpha and delta were isolated from S. pombe. Both pol alpha+ and pol delta+ genes are single copy genes in haploid cells and are essential for cell viability. In contrast to Saccharomyces cerevisiae homologs, the steady-state transcripts of both S. pombe pol alpha+ and pol delta+ genes were present throughout the cell cycle. Sequence analysis of the pol alpha+ and pol delta+ genes did not reveal the Mlu I motifs in their upstream sequences that are involved in cell cycle-dependent transcription of S. cerevisiae DNA synthesis genes as well as the S. pombe cdc22+ gene at the G1/S boundary. However, five near-match Mlu I motifs were found in the upstream region of the pol alpha+ gene. S. pombe DNA polymerases alpha and delta proteins were also expressed constantly throughout the cell cycle. In addition, the enzymatic activity of the S. pombe DNA polymerase alpha measured by in vitro assay was detected at all stages of the cell cycle. Thus, these S. pombe replicative DNA polymerases, like that of S. pombe cdc17+ gene, are expressed throughout the cell cycle at the transcriptional and protein level. These results indicate that S. pombe has at least two regulatory modes for the expression of genes involved in DNA replication and DNA precursor synthesis.


SUMO targeting of a stress-tolerant Ulp1 SUMO protease.
SUMO proteases of the SENP/Ulp family are master regulators of both sumoylation and desumoylation and regulate SUMO homeostasis in eukaryotic cells. SUMO conjugates rapidly increase in response to cellular stress, including nutrient starvation, hypoxia, osmotic stress, DNA damage, heat shock, and other proteotoxic stressors. Nevertheless, little is known about the regulation and targeting of SUMO proteases during stress. To this end we have undertaken a detailed comparison of the SUMO-binding activity of the budding yeast protein Ulp1 (ScUlp1) and its ortholog in the thermotolerant yeast Kluyveromyces marxianus, KmUlp1. We find that the catalytic UD domains of both ScUlp1 and KmUlp1 show a high degree of sequence conservation, complement a ulp1Δ mutant in vivo, and process a SUMO precursor in vitro. Next, to compare the SUMO-trapping features of both SUMO proteases we produced catalytically inactive recombinant fragments of the UD domains of ScUlp1 and KmUlp1, termed ScUTAG and KmUTAG respectively. Both ScUTAG and KmUTAG were able to efficiently bind a variety of purified SUMO isoforms and bound immobilized SUMO1 with nanomolar affinity. However, KmUTAG showed a greatly enhanced ability to bind SUMO and SUMO-modified proteins in the presence of oxidative, temperature and other stressors that induce protein misfolding. We also investigated whether a SUMO-interacting motif (SIM) in the UD domain of KmULP1 that is not conserved in ScUlp1 may contribute to the SUMO-binding properties of KmUTAG. In summary, our data reveal important details about how SUMO proteases target and bind their sumoylated substrates, especially under stress conditions. We also show that the robust pan-SUMO binding features of KmUTAG can be exploited to detect and study SUMO-modified proteins in cell culture systems.


ERBB2, TBX2, RPS6KB1, and MYC alterations in breast tissues of BRCA1 and BRCA2 mutation carriers.
Breast cancer risk is greatly increased in women who carry mutations in the BRCA1 or BRCA2 genes. Because breast cancer initiation is different between BRCA1/2 mutation carriers and women who do not carry mutations, it is possible that the mechanism of breast cancer progression is also different. Histopathologic and genetic studies have supported this hypothesis. To test this hypothesis further, we utilized a large cohort of women who underwent therapeutic mastectomy (TM) and contralateral prophylactic mastectomy (PM). From this cohort, we developed case groups of women with a family history of breast cancer with BRCA1/2 deleterious mutations, with unclassified variant alterations, and with no detected mutation and matched these cases with sporadic controls from the same TM and PM cohort. Fluorescence in situ hybridization was performed on paraffin sections by use of dual-color probes for ERBB2/CEP17, MYC/CEP8, TBX2/CEP17, and RPS6KB1/CEP17. All malignant and benign lesions, including putative precursor lesions, were studied. The invasive cancers from deleterious mutation carriers had a higher prevalence of duplication of MYC (P = 0.006) and TBX2 (P = 0.0008) compared to controls and a lower prevalence of ERBB2 amplification (P = 0.011). Coduplication of MYC and TBX2 was common in the in situ and invasive lesions from the deleterious mutation carriers. The odds ratio of having a BRCA1/2 mutation is 31.4 (95% CI = 1.7-569) when MYC and TBX2 are coduplicated but ERBB2 is normal. Unclassified variant carriers/no mutation detected and sporadic controls had a similar prevalence of alterations, suggesting that hereditary patients with no deleterious mutations follow a progression pathway similar to that of sporadic cases. With the exception of one atypical ductal hyperplasia lesion, no putative precursor lesion showed any detectable alteration of the probes tested. There was no significant intratumoral heterogeneity of genetic alterations. Our data confirm that a specific pattern of genomic instability characterizes BRCA1/2-related cancers and that this pattern has implications for the biology of these cancers. Moreover, our current and previous results emphasize the interaction between phenotype and genotype in BRCA1/2-related breast cancers and that a combination of morphologic features and alterations of ERBB2, MYC, and TBX2 may better define mechanisms of tumor progression, as well as determine which patients are more likely to carry BRCA1/2 mutations.


A full-length cDNA of hREV3 is predicted to encode DNA polymerase zeta for damage-induced mutagenesis in humans.
DNA damage can cause mutations which in turn may lead to carcinogenesis. In the yeast Saccharomyces cerevisiae, DNA damage-induced mutagenesis pathway requires the REV3 gene. It encodes the catalytic subunit of DNA polymerase zeta that specifically functions in translesion DNA synthesis. We have cloned a cDNA of the human homologue of REV3 (hREV3), which consists of 10,716 bp and codes for a protein of 3130 amino acid residues (352,737 Da). Its C-terminal 755 amino acids show extensive homology with the yeast protein at the C-terminus: 43% identity and 74% similarity. This region contains the six highly conserved DNA polymerase motifs. Furthermore, we have identified four sequence motifs in the N-terminal region outside the polymerase domain that are conserved in DNA polymerase delta from various sources. Three of which are present in DNA polymerase zeta encoded by human, yeast, and plant REV3 genes, indicating that this protein is a member of the DNA polymerase delta family. DNA polymerases delta and zeta are structurally distinguished by the presence of a specific delta IV motif in the former and motifs zeta I and zeta II in the latter, respectively. Human DNA polymerase zeta is ubiquitously expressed in various tissues, consistent with the notion that the hREV3 pathway may be a fundamental mechanism of damage-induced mutagenesis in humans.


Developing future medical educators in an Australian medical program: supervisors' reflections on the first four years of MD Professional Project implementation.
Increasingly, professional bodies expect doctors to not only provide patient care but also educate students, trainees and patients. Few medical students, however, receive formal tuition in terms of the theory and practice of medical education. A curriculum restructure from an MBBS to a Doctor of Medicine (MD) program provided an opportunity to develop three Masters streams: Clinical research, Capstones and educational Professional Projects. This submission describes how one Australian medical school is preparing some students for their future roles as medical educators through MD Professional Projects. Framed by the 12 roles of the medical 'teacher', most students undertaking these projects take on Resource Developer (including simulation) and Assessor roles. For those choosing resource development (excluding simulation) or assessment, the Association for Medical Education (AMEE) Student Essential Skills in Medical Education (ESME) Course is compulsory. For those choosing educational research, the ESME Course is optional. By December 2020, four MD cohorts will have graduated with 69 students having undertaken educational MD Professional Projects, with fifty-one completing the ESME Course. MD students have created a range of resources for the curriculum, their colleagues and the local healthcare community. In addition to the expected learning we identified additional value-added outcomes for learners (e.g. skill development), the curriculum (e.g. areas of difficulty), academic supervisors' roles (e.g. role-modelling) and for the health care community (e.g. as expert reviewers). Engaging in scholarly activities such the ESME Course and developing learning resources not only provided MD students with a more in-depth theoretical knowledge in a range of clinical areas, but also developed skills that would prepare them for their future roles as medical educators. As supervisors, we identified the value these projects add to the broader health community as well as personal and professional benefits for ourselves.


NADPH Oxidase Overactivity Underlies Telomere Shortening in Human Atherosclerosis.
Telomere shortening and oxidative stress are involved in the pathogenesis of atherosclerosis. Different studies have shown that phagocytic NADPH oxidase is associated with this disease. This study aimed to investigate the association between phagocytic NADPH oxidase and telomere shortening in human atherosclerosis. To assess this potential association, telomere length and phagocytic NADPH oxidase activity were determined by PCR and chemiluminescence, respectively, in a population of asymptomatic subjects free of overt clinical atherosclerosis. We also measured serum 8-hydroxy-2-deoxyguanosine (8-OHdG) levels (an index of oxidative stress) and carotid intima-media thickness (IMT), a surrogate marker of atherosclerosis. After adjusting them for age and sex, telomere length inversely correlated (p < 0.05) with NADPH oxidase-mediated superoxide production, with 8-OHdG values, and with carotid IMT. Interestingly, the asymptomatic subjects with plaques have a lower telomere length (p < 0.05), and higher values of plasma 8-OHdG and superoxide production (p < 0.05). These data were confirmed in a second population in which patients with coronary artery disease showed lower telomere length and higher 8-OHdG and superoxide production than the asymptomatic subjects. In both studies, NADPH oxidase-dependent superoxide production in phagocytic cells was only due to the specific expression of the Nox2 isoform. In conclusion, these findings suggest that phagocytic NADPH oxidase may be involved in oxidative stress-mediated telomere shortening, and that this axis may be critically involved in human atherosclerosis.


Barriers to exercise prescription and participation in people with mental illness: the perspectives of nurses working in mental health.
Exercise is valuable in the treatment of mental illness, yet personal and organizational barriers limit widespread implementation by nurses in mental health settings. Using a self-report questionnaire, we sought to identify how often nurses prescribe exercise and their level of agreement with previously identified barriers to exercise prescription and participation for mental health consumers. Nurses disagree that many of the previously identified barriers should impede exercise prescription for people with mental illness. Nurses agree that many of the barriers expressed by mental health consumers might prevent exercise participation. Our study provides valuable new insight into the role of nurses in the provision of exercise for people with mental illness; however, it is limited to a small sample. Confirmation of these findings in larger, geographically and professionally diverse groups is needed. Evidence is mounting for the efficacy of exercise in the treatment of people with mental illness. Nurses working in mental health settings are well placed to provide exercise advice for people with mental illness. However, quantitative examinations of the barriers to exercise prescription experienced by nurses, or their views regarding the barriers to exercise participation experienced by people with mental illness, are lacking. In this study, 34 nurses completed the Exercise in Mental Illness Questionnaire-Health Professionals Version (EMIQ-HP). This survey examined the frequency of exercise prescription and the level of agreement with statements regarding barriers to exercise prescription for, and exercise participation by, people with mental illness. The level of agreement scores for statements for each section was summed, with a higher score indicating a higher level of agreement. Nurses disagree with many of the barriers to exercise prescription presented in the literature. The level of agreement scores did not differ between nurses who prescribe exercise 'Always', 'Most of the time', 'Occasionally' or 'Never'. We found a non-significant negative relationship between frequency of exercise prescription and summed level of agreement scores for barriers to exercise prescription. Consensus regarding barriers to exercise participation by mental health consumers is less clear. This study provides valuable new insight into the role of nurses in the provision of exercise for people with mental illness. Confirmation in larger samples is needed before translation of research to practice.


Effect of dehydration on gastrointestinal function at rest and during exercise in humans.
Dehydration leads to the aggravation of gastrointestinal (GI) complaints during exercise. The aim of this study was to examine the effect of dehydration on various GI parameters during strenuous exercise. Ten healthy well-trained men were investigated in dehydrated and in euhydrated conditions. Dehydration took place before the experiments using a dehydration regimen in a sauna leading to a 3% loss of body mass. Each experiment consisted of 1 h pre-exercise rest, 1.5 h cycling at 70% maximal exercise intensity, and 3.5 h post-exercise rest. During cycling, liquid gastric emptying (GE), orocaecal transit time (OCTT) and intestinal permeability and glucose absorption were measured. The GI-symptoms were scored using a questionnaire. Body temperature, plasma volume and vasopressin were measured before and after cycling. The GE was significantly slower during dehydration [median time to peak 13C enrichment in the breath sample (13C-TTP) 23.6 min, range 13.7-50.0 min, P = 0.02] than in the control situation (median 13C-TTP 17.1 min, range 9.8-38.4 min). The OCTT was unchanged (median 173 min, range 98-263 min compared to median 128 min, range 98-195 min, P = 0.18). Dehydration did not change intestinal permeability, glucose absorption, plasma volume, rectal temperature or plasma vasopressin concentration. In the dehydration experiment, exercise induced a significant increase in nausea (P = 0.01) and epigastric cramps (P = 0.05), in contrast to the control situation. In both experiments, exercise led to a significant increase in rectal temperature and plasma vasopressin concentration, and a significant decrease in plasma volume. The increase in plasma vasopressin concentration was significantly higher in the dehydration experiment (P = 0.015). No significant differences in either the post-exercise rectal temperatures or in plasma volumes was observed. The difference in GE between the two experiments was significantly correlated with the difference in nausea score (r = 0.87, P = 0.002). We concluded that dehydration leads to a delayed GE but not to differences in OCTT, intestinal permeability or glucose uptake during intense cycling. The delay in GE is significantly associated with an increase in exercise-induced nausea.


Examining the pathways linking chronic sleep restriction to obesity.
A growing number of studies have identified chronic sleep restriction as a potential risk factor for obesity. This could have important implications for how obesity is prevented and managed, but current understanding of the processes linking chronic sleep restriction to obesity is incomplete. In this paper, we examined some of the pathways that could underlie the relationship between chronic sleep restriction and obesity. This involved exploring some of the potential environmental, health, behavioral, and sociodemographic determinants of chronic sleep restriction, which require further investigation in this context. Three pathways that could potentially link chronic sleep restriction to obesity were then examined: (1) altered neuroendocrine and metabolic function, (2) impaired glucose regulation, and (3) waking behavior. The selected pathways linking chronic sleep restriction to obesity reviewed in this paper are presented in a schematic representation; this may be used to guide future research in this area. This area of research is important because it may lead to more effective interventions and strategies to combat the present obesity epidemic.


Variations in genome-wide gene expression in identical twins - a study of primary osteoblast-like culture from female twins discordant for osteoporosis.
Monozygotic twin pairs who are genetically identical would be potentially useful in gene expression study for specific traits as cases and controls, because there would be much less gene expression variation within pairs compared to two unrelated individuals. However the twin pair has to be discordant for the particular trait or phenotype excluding those resulting from known confounders. Such discordant monozygotic twin pairs are rare and very few studies have explored the potential usefulness of this approach. We studied genome-wide gene expression in primary osteoblast-like culture from marrow aspirates obtained from three pairs of monozygotic twins. We used the latest Affymetrix microchip contains probe sets for more than 20,000 genes. Two pairs were discordant for bone mineral density at the hip by more than one standard deviation, and the third pair was unrelated concordant and used as control. Only 1.5% on average of genes showed variation in expression within pairs as compared to 5% between pairs or over 15% from the literature. Importantly we identified several groups of genes showing variations within the discordant pairs and not within the concordant pair such as chondroitin beta 1,4 N-acetylgalactosaminyltransferase, inhibin beta A, interleukin 1 beta and colony stimulating factor 1 macrophage. These genes are known to have potential roles in bone physiology relating to bone density, osteoporosis and osteoarthritis. Using the example of osteoblast-like cells in our monozygotic discordant twins for osteoporosis, we identified genes showing differential expression. Although without further experiment, we cannot confirm or conclude these are genes definitely related to bone physiology, we believe we have shown the potential and cost-effectiveness of further gene expression studies in discordant monozygotic twin pairs. A replication study for confirmation is essential.


Visceral tissue mass and rumen volume in dairy cows during the transition from late gestation to early lactation.
The objectives were to measure the effects of transition and supplemental barley or rumen-protected protein on visceral tissue mass in dairy cows and the effects of transition and barley on rumen volume and liquid turnover. Cows were individually fed a grass silage-based gestation ration to meet energy and protein requirements for body weight stasis beginning 6 wk before expected calving. A corn silage-based lactation ration was individually fed ad libitum after calving. In the visceral mass study, 36 cows were randomly assigned to one of 3 dietary treatments: basal ration or basal ration plus either 800 g dry matter (DM) of barley meal per day or 750 g DM of rumen-protected soybean protein per day. Cows were slaughtered at 21 and 7 d before expected calving date or at 10 and 22 d postpartum. Visceral mass and rumen papillae characteristics were measured. Diets had little effect on visceral mass. The mass of the reticulo-rumen, small intestine, large intestine, and liver was, or tended to be, greater at 22 d postpartum but not at 10 d postpartum before DM intake had increased. Rumen papillae mass increased at 10 d postpartum, perhaps in response to increased concentrates. Mesenteric fat decreased after calving, reflecting body fat mobilization. Ten rumen-cannulated cows were fed the basal gestation ration alone or supplemented with 880 g of barley meal DM. Rumen volumes and liquid dilution rates were measured at 17 and 8 d before calving and at 10, 20, and 31 d postpartum. Feeding barley had no effects. After calving, rumen DM volume and liquid dilution rate increased, but liquid volume did not increase. Changes in gastrointestinal and liver mass during transition were apparently a consequence of changes in DM intake and nutrient supply and not initiation of lactation per se.


Intraoperative visualization of fiber tracking based reconstruction of language pathways in glioma surgery.
For neuroepithelial tumors, the surgical goal is maximum resection with preservation of neurological function. This is contributed to by intraoperative magnetic resonance imaging (iMRI) combined with multimodal navigation. We evaluated the contribution of diffusion tensor imaging (DTI)-based fiber tracking of language pathways with 2 different algorithms (tensor deflection, connectivity analysis [CA]) integrated in the navigation on the surgical outcome. We evaluated 32 patients with neuroepithelial tumors who underwent surgery with DTI-based fiber tracking of language pathways integrated in neuronavigation. The tensor deflection algorithm was routinely used and its results intraoperatively displayed in all cases. The CA algorithm was furthermore evaluated in 23 cases. Volumetric assessment was performed in pre- and intraoperative MR images. To evaluate the benefit of fiber tractography, language deficits were evaluated pre- and postoperatively and compared with the volumetric analysis. Final gross-total resection was performed in 40.6% of patients. Absolute tumor volume was reduced from 55.33 ± 63.77 cm(3) to 20.61 ± 21.67 cm(3) in first iMRI resection control, to finally 11.56 ± 21.92 cm(3) (P < .01). Fiber tracking of the 2 algorithms showed a deviation of the displayed 3D objects by <5 mm. In long-term follow-up only 1 patient (3.1%) had a persistent language deficit. Intraoperative visualization of language-related cortical areas and the connecting pathways with DTI-based fiber tracking can be successfully performed and integrated in the navigation system. In a setting of intraoperative high-field MRI this contributes to maximum tumor resection with low postoperative morbidity.


Natural killer T cells accelerate atherogenesis in mice.
We have investigated the potential role of CD1d-restricted natural killer T (NKT) cells in the development of atherosclerosis in mice. When fed an atherogenic diet (AD), NKT cell-deficient CD1d(-/-) mice had significantly smaller atherosclerotic lesions than AD-fed C57BL/6 (wild-type [WT]) mice. A significant reduction in atherosclerotic lesions was also demonstrated in AD-fed, low-density lipoprotein receptor-deficient (Ldlr(-/-)) mice reconstituted with CD1d(-/-) bone marrow cells compared with the lesions observed in Ldlr(-/-)mice reconstituted with WT marrow cells. In addition, repeated injections of alpha-GalCer or the related glycolipid OCH to apolipoprotein E knockout (apoE(-/-)) mice during the early phase of atherosclerosis significantly enlarged the lesion areas compared with mice injected with vehicle control. However, administering alpha-GalCer to apoE(-/-) mice with established lesions did not significantly increase the lesion area but considerably decreased the collagen content. Atherosclerosis development in either AD-fed WT or apoE(-/-) mice was associated with the presence of Valpha14Jalpha18 transcripts in the atherosclerotic arterial walls, indicating that NKT cells were recruited to these lesions. Thioglycolate-elicited macrophages pulsed with oxidized low-density lipoproteins expressed enhanced CD1d levels and induced NKT cells to produce interferon-gamma, a potentially proatherogenic T-helper 1 (TH1) cytokine. Collectively, we conclude that NKT cells are proatherogenic in mice.


Tumor markers in prostate cancer I: blood-based markers.
The introduction of total prostate specific antigen (total PSA) testing in blood has revolutionized the detection and management of men with prostate cancer (PCa). The objective of this review was to discuss the challenges of PCa biomarker research, definition of the type of PCa biomarkers, the statistical considerations for biomarker discovery and validation, and to review the literature regarding total PSA velocity and novel blood-based biomarkers. An English-language literature review of the Medline database (1990 to August 2010) of published data on blood-based biomarkers and PCa was undertaken. The inherent biological variability of total PSA levels affects the interpretation of any single result. Men who will eventually develop PCa have increased total PSA levels years or decades before the cancer is diagnosed. Total PSA velocity improves predictiveness of total PSA only marginally, limiting its value for PCa screening and prognostication. The combination of PSA molecular forms and other biomarkers improve PCa detection substantially. Several novel blood-based biomarkers such as human glandular kallikrein 2 (hK2), urokinase plasminogen activator (uPA) and its receptor (uPAR), transforming growth factor-beta 1 (TGF-β1); interleukin-6 (IL-6) and its receptor (IL-6R) may help PCa diagnosis, staging, prognostication, and monitoring. Panels of biomarkers that capture the biologic potential of PCa are in the process of being validated for PCa prognostication. PSA is a strong prognostic marker for long-term risk of clinically relevant cancer. However, there is a need for novel biomarkers that aid clinical decision making about biopsy and initial treatment. There is no doubt that progress will continue based on the integrated collaboration of researchers, clinicians and biomedical firms.


Complications of Radiotherapy and Radiosurgery in the Brain and Spine.
Radiation therapy is an integral part of the standard of care for many patients with brain and spine tumors. Stereotactic radiation surgery is increasingly being used as an adjuvant therapy as well as a sole treatment. However, despite newer and more focused techniques, radiation therapy still causes significant neurotoxicity. In this article, we reviewed the scientific literature, presented cases of patients who had developed different complications related to conventional radiation therapy or radiosurgery (gamma knife), demonstrated the imaging findings, and discussed the relevant clinical information for the correct diagnoses. Radiation therapy can cause injury in different ways: directly damaging the structures included in the radiation portal, indirectly affecting the blood vessels, and increasing the chance of tumor development. We also divided radiation complications according to the time of occurrence: acute (0 to 4 weeks), early delayed (4 weeks to months), and late delayed (months to years). With the increasing application of radiation therapy for the treatment of CNS tumors, it is important for the neuroradiologist to recognize the many possible complications of radiation therapy. Although this may cause significant diagnostic challenges, understanding the pathophysiology, time course of onset, and imaging features may help institute early therapy and prevent possible deleterious outcomes. To recognize the main complications of radiation therapy and stereotactic radiosurgery in the brain and spine, and to highlight the imaging findings to improve the diagnostic process and treatment planning.


Transient Receptor Potential Canonical 3 (TRPC3) Channels Are Required for Hypothalamic Glucose Detection and Energy Homeostasis.
The mediobasal hypothalamus (MBH) contains neurons capable of directly detecting metabolic signals such as glucose to control energy homeostasis. Among them, glucose-excited (GE) neurons increase their electrical activity when glucose rises. In view of previous work, we hypothesized that transient receptor potential canonical type 3 (TRPC3) channels are involved in hypothalamic glucose detection and the control of energy homeostasis. To investigate the role of TRPC3, we used constitutive and conditional TRPC3-deficient mouse models. Hypothalamic glucose detection was studied in vivo by measuring food intake and insulin secretion in response to increased brain glucose level. The role of TRPC3 in GE neuron response to glucose was studied by using in vitro calcium imaging on freshly dissociated MBH neurons. We found that whole-body and MBH TRPC3-deficient mice have increased body weight and food intake. The anorectic effect of intracerebroventricular glucose and the insulin secretory response to intracarotid glucose injection are blunted in TRPC3-deficient mice. TRPC3 loss of function or pharmacological inhibition blunts calcium responses to glucose in MBH neurons in vitro. Together, the results demonstrate that TRPC3 channels are required for the response to glucose of MBH GE neurons and the central effect of glucose on insulin secretion and food intake.


Prospective association between added sugars and frailty in older adults.
Sugar-sweetened beverages and added sugars (monosaccharides and disaccharides) in the diet are associated with obesity, diabetes, and cardiovascular disease, which are all risk factors for decline in physical function among older adults. The aim of this study was to examine the association between added sugars in the diet and incidence of frailty in older people. Data were taken from 1973 Spanish adults ≥60 y old from the Seniors-ENRICA cohort. In 2008-2010 (baseline), consumption of added sugars (including those in fruit juices) was obtained using a validated diet history. Study participants were followed up until 2012-2013 to assess frailty based on Fried's criteria. Statistical analyses were performed with logistic regression adjusted for age, sex, education, smoking status, body mass index, energy intake, self-reported comorbidities, Mediterranean Diet Adherence Score (excluding sweetened drinks and pastries), TV watching time, and leisure-time physical activity. Compared with participants consuming <15 g/d added sugars (lowest tertile), those consuming ≥36 g/d (highest tertile) were more likely to develop frailty (OR: 2.27; 95% CI: 1.34, 3.90; P-trend = 0.003). The frailty components "low physical activity" and "unintentional weight loss" increased dose dependently with added sugars. Association with frailty was strongest for sugars added during food production. Intake of sugars naturally appearing in foods was not associated with frailty. The consumption of added sugars in the diet of older people was associated with frailty, mainly when present in processed foods. The frailty components that were most closely associated with added sugars were low level of physical activity and unintentional weight loss. Future research should determine whether there is a causal relation between added sugars and frailty.


Immunoglobulin hypermutation in cultured cells.
Studies of endogenous and engineered Ig genes in mice have begun to reveal some of the cis-acting regions that are involved in the somatic hypermutation of variable regions in vivo. These studies suggest that the initiation of transcription plays a role in this process. However, it will be difficult to identify and manipulate the individual genetic elements and the trans-acting proteins that regulate and target the mutational events using solely in vivo assays. These studies would be greatly facilitated if constructs containing the genetic elements that are essential for V-region mutation could be transfected into cultured cells and undergo high rates of V-region mutation in vitro, and if permissive and non-permissive cell lines could be identified. Such in vitro systems would also allow a detailed molecular and biochemical analysis of this process. Here, we discuss some of the in vitro systems that have been developed and use data from our own studies in cultured cells to illustrate the potential benefits of studying V-region hypermutation in model in vitro systems.


Computational design of nucleic acid feedback control circuits.
The design of synthetic circuits for controlling molecular-scale processes is an important goal of synthetic biology, with potential applications in future in vitro and in vivo biotechnology. In this paper, we present a computational approach for designing feedback control circuits constructed from nucleic acids. Our approach relies on an existing methodology for expressing signal processing and control circuits as biomolecular reactions. We first extend the methodology so that circuits can be expressed using just two classes of reactions: catalysis and annihilation. We then propose implementations of these reactions in three distinct classes of nucleic acid circuits, which rely on DNA strand displacement, DNA enzyme and RNA enzyme mechanisms, respectively. We use these implementations to design a Proportional Integral controller, capable of regulating the output of a system according to a given reference signal, and discuss the trade-offs between the different approaches. As a proof of principle, we implement our methodology as an extension to a DNA strand displacement software tool, thus allowing a broad range of nucleic acid circuits to be designed and analyzed within a common modeling framework.


Access to US primary care physicians for new patients concerned about smoking or weight.
Tobacco smoking and obesity are leading causes of preventable morbidity and mortality in the US, and primary care physicians are the main source of preventive care. However, it is not known whether access for new patients is affected by an expression of interest in preventive care. In a 2015 audit, we called US primary care physicians' offices to request appointment information regarding new patient physicals for simulated patients. Simulated patients were differentiated by smoking concerns (N = 907), weight concerns (N = 867), or no health concerns ("healthy" patients; N = 3561). Additionally, patient profiles varied by race/ethnicity, sex, and insurance type. We also examined whether access differed in states that expanded Medicaid under the Affordable Care Act. We found that physicians' offices were no more likely to offer appointments to patients with smoking concerns than to healthy patients (54% vs. 55%; p-value = 0.56), and patients with smoking concerns were offered fewer appointments than patients with weight concerns (54% vs. 62%, p-value < 0.01). In analyses adjusted for covariates, smoking concerns did not improve appointment offers for any patient group, and reduced Medicare patients' offers in Medicaid expansion states by 9 percentage points relative to healthy patients (95% CI: -16, -2). Health concerns did not statistically significantly affect waits-to-appointment. Our results suggest that patients with smoking concerns are no more likely to be offered new patient appointments than those with no health concerns. The greater likelihood of appointment offers for some patients with weight concerns is encouraging for obesity prevention and management.


Assessing potential health risks to fish and humans using mercury concentrations in inland fish from across western Canada and the United States.
Fish represent high quality protein and nutrient sources, but Hg contamination is ubiquitous in aquatic ecosystems and can pose health risks to fish and their consumers. Potential health risks posed to fish and humans by Hg contamination in fish were assessed in western Canada and the United States. A large compilation of inland fish Hg concentrations was evaluated in terms of potential health risk to the fish themselves, health risk to predatory fish that consume Hg contaminated fish, and to humans that consume Hg contaminated fish. The probability that a fish collected from a given location would exceed a Hg concentration benchmark relevant to a health risk was calculated. These exceedance probabilities and their associated uncertainties were characterized for fish of multiple size classes at multiple health-relevant benchmarks. The approach was novel and allowed for the assessment of the potential for deleterious health effects in fish and humans associated with Hg contamination in fish across this broad study area. Exceedance probabilities were relatively common at low Hg concentration benchmarks, particularly for fish in larger size classes. Specifically, median exceedances for the largest size classes of fish evaluated at the lowest Hg concentration benchmarks were 0.73 (potential health risks to fish themselves), 0.90 (potential health risk to predatory fish that consume Hg contaminated fish), and 0.97 (potential for restricted fish consumption by humans), but diminished to essentially zero at the highest benchmarks and smallest fish size classes. Exceedances of benchmarks are likely to have deleterious health effects on fish and limit recommended amounts of fish humans consume in western Canada and the United States. Results presented here are not intended to subvert or replace local fish Hg data or consumption advice, but provide a basis for identifying areas of potential health risk and developing more focused future research and monitoring efforts.


Tuberculosis: progress and advances in development of new drugs, treatment regimens, and host-directed therapies.
Tuberculosis remains the world's leading cause of death from an infectious disease, responsible for an estimated 1 674 000 deaths annually. WHO estimated 600 000 cases of rifampicin-resistant tuberculosis in 2016-of which 490 000 were multidrug resistant (MDR), with less than 50% survival after receiving recommended treatment regimens. Concerted efforts of stakeholders, advocates, and researchers are advancing further development of shorter course, more effective, safer, and better tolerated treatment regimens. We review the developmental pipeline and landscape of new and repurposed tuberculosis drugs, treatment regimens, and host-directed therapies (HDTs) for drug-sensitive and drug-resistant tuberculosis. 14 candidate drugs for drug-susceptible, drug-resistant, and latent tuberculosis are in clinical stages of drug development; nine are novel in phase 1 and 2 trials, and three new drugs are in advanced stages of development for MDR tuberculosis. Specific updates are provided on clinical trials of bedaquiline, delamanid, pretomanid, and other licensed or repurposed drugs that are undergoing investigation, including trials aimed at shortening duration of tuberculosis treatment, improving treatment outcomes and patient adherence, and reducing toxic effects. Ongoing clinical trials for shortening tuberculosis treatment duration, improving treatment outcomes in MDR tuberculosis, and preventing disease in people with latent tuberculosis infection are reviewed. A range of HDTs and immune-based treatments are under investigation as adjunctive therapy for shortening duration of therapy, preventing permanent lung injury, and improving treatment outcomes of MDR tuberculosis. We discuss the HDT development pipeline, ongoing clinical trials, and translational research efforts for adjunct tuberculosis treatment.


Uterine function in the mouse requires speckle-type poz protein.
Speckle-type poz protein (SPOP) is an E3-ubiquitin ligase adaptor for turnover of a diverse number of proteins involved in key cellular processes such as chromatin remodeling, transcriptional regulation, and cell signaling. Genomic analysis revealed that SPOP somatic mutations are found in a subset of endometrial cancers, suggesting that these mutations act as oncogenic drivers of this gynecologic malignancy. These studies also raise the question as to the role of wild-type SPOP in normal uterine function. To address this question, we generated a mouse model (Spopd/d) in which SPOP is ablated in uterine cells that express the PGR. Fertility studies demonstrated that SPOP is required for embryo implantation and for endometrial decidualization. Molecular analysis revealed that expression levels of the PGR at the protein and transcript level are significantly reduced in the Spopd/d uterus. While this result was unexpected, this finding explains in part the dysfunctional phenotype of the Spopd/d uterus. Moderate increased levels of the ESR1, GATA2, and SRC2 were detected in the Spopd/d uterus, suggesting that SPOP is required to maintain the proteome for normal uterine function. With age, the Spopd/d endometrium exhibits large glandular cysts with foci of epithelial proliferation, further supporting a role for SPOP in maintaining a healthy uterus. Collectively, studies on the Spopd/d mouse support an important role for SPOP in normal uterine function and suggest that this mouse model may prove useful to study the role of SPOP-loss-of-function mutations in the etiopathogenesis of endometrial cancer.


Prediction of concentration-time profile and its inter-individual variability following the dermal drug absorption.
Estimation of systemic exposure after absorption of any xenobiotic from the skin is very important in development of dermal pharmaceutical products as well as assessing un-intended exposures due to cosmetic products or environmental and occupational compounds. Historically, animal models have been used to evaluate dermal drug absorption before conducting human trials. However, occasional disparity between the animal and human data plus rising public interest and regulatory requirements to reduce animal usage in research combined with high cost and time-consuming attributes of animal experiments have prompted many academic and industrial researchers to develop economically viable and scientifically robust in silico and in vitro methods to assess dermal drug absorption. There are a number of in silico models available in literature from quantitative structure-activity relationship to semi-mechanistic to physiologically based mechanistic models. Nonetheless, to the best of our knowledge, so far, there has been no attempt to combine mechanistic skin absorption model with database of physiological variability to simulate the inter- and intra-individual variability observed in human trials. Thus, we report here mechanistic dermal absorption model with formulation, stratum corneum, viable epidermis-dermis and blood compartments along with datab"ase of human dermal physiological variability including gender, ethnic and site of application variations. The developed model is incorporated into the Simcyp simulator which is a 'bottom-up' platform and database for mechanistic modelling and simulation of the drug disposition process using full body physiologically based pharmacokinetics model. The built model is validated using the clinical pharmacokinetic data from five different topical formulations of diclofenac. The effect of penetration enhancers, site of application, gender and ethnic variations were incorporated to simulate the clinical trials. The applied mechanistic dermal absorption model when combined with skin physiological database was able to recover well the observed clinical pharmacokinetics and population variability in all the five validation studies.


A factor VIII-nanobody fusion protein forming an ultrastable complex with VWF: effect on clearance and antibody formation.
Von Willebrand factor (VWF) modulates factor VIII (FVIII) clearance and the anti-FVIII immune response. Despite the high affinity that defines the FVIII/VWF interaction, association/dissociation kinetics dictates 2% to 5% FVIII being present as free protein. To avoid free FVIII when studying the FVIII-VWF complex in vivo, we designed a FVIII-nanobody fusion protein, with the nanobody part being directed against VWF. This fusion protein, designated FVIII-KB013bv, had a 25-fold higher affinity compared with B-domainless FVIII (BDD-FVIII) for VWF. In vitro analysis revealed full cofactor activity in 1-stage clotting and chromogenic assays (activity/antigen ratio 1.0 ± 0.3 and 1.1 ± 0.3, respectively). In vivo, FVIII-013bv displayed a twofold increased mean residence time compared with BDD-FVIII (3.0 hours vs 1.6 hours). In a tail clip-bleeding assay performed 24 hours after FVIII infusion, blood loss was significantly reduced in mice receiving FVIII-KB013bv vs BDD-FVIII (15 ± 7 μL vs 194 ± 146 μL; P = .0043). Unexpectedly, when examining anti-FVIII antibody formation in FVIII-deficient mice, the immune-response toward FVIII-KB013bv was significantly reduced compared with BDD-FVIII (1/8 vs 14/16 mice produced anti-FVIII antibodies after treatment with FVIII-KB013bv and BDD-FVIII, respectively). Our data show that a stabilized interaction between FVIII and VWF is associated with a prolonged survival of FVIII and a reduced immune response against FVIII.


Cross-linguistic interactions influence reading development in bilinguals: a comparison between early balanced French-Basque and Spanish-Basque bilingual children.
This study investigates whether orthographic consistency and transparency of languages have an impact on the development of reading strategies and reading sub-skills (i.e. phonemic awareness and visual attention span) in bilingual children. We evaluated 21 French (opaque)-Basque (transparent) bilingual children and 21 Spanish (transparent)-Basque (transparent) bilingual children at Grade 2, and 16 additional children of each group at Grade 5. All of them were assessed in their common language (i.e. Basque) on tasks measuring word and pseudoword reading, phonemic awareness and visual attention span skills. The Spanish speaking groups showed better Basque pseudoword reading and better phonemic awareness abilities than their French speaking peers, but only in the most difficult conditions of the tasks. However, on the visual attention span task, the French-Basque bilinguals showed the most efficient visual processing strategies to perform the task. Therefore, learning to read in an additional language affected differently Basque literacy skills, depending on whether this additional orthography was opaque (e.g. French) or transparent (e.g. Spanish). Moreover, we showed that the most noteworthy effects of Spanish and French orthographic transparency on Basque performance were related to the size of the phonological and visual grain used to perform the tasks.