Discrimination and identification of lexical tones and consonants in Mandarin-speaking children using cochlear implants.
Mandarin-speaking adults using cochlear implants (CI) experience more difficulties in perceiving lexical tones than consonants. This problem may result from the fact that CIs provide relatively sufficient temporal envelope information for consonant perception in quiet environments, but do not convey the fine spectro-temporal information considered to be necessary for accurate pitch perception. Another possibility is that Mandarin speakers with post-lingual hearing loss have developed language-specific use of these acoustic cues, impeding lexical tone processing under CI conditions. To investigate this latter hypothesis, syllable discrimination and word identification abilities for Mandarin consonants (place and manner) and lexical-tone contrasts (tones 1 vs 3 and 1 vs 2) were measured in 15 Mandarin-speaking children using CIs and age-matched children with normal hearing (NH). In the discrimination task, only children using CIs exhibited significantly lower scores for consonant place contrasts compared to other contrasts, including lexical tones. In the word identification task, children using CIs showed lower performance for all contrasts compared to children with NH, but they both showed specific difficulties with tone 1 vs 2 contrasts. This study suggests that Mandarin-speaking children using CIs are able to discriminate and identify lexical tones and, perhaps more surprisingly, have more difficulties when discriminating consonants.


The quick wins paradox.
Many leaders taking on new roles try to prove themselves early on by going after quick wins--fresh, visible contributions to the business. But in the pursuit of early results, those leaders often fall into traps that prevent them from benefiting from their achievements. To succeed in their new positions, leaders must realize that the teams they have inherited are also experiencing change. Instead of focusing on an individual accomplishment, leaders need to work with team members on a collective quick win. In a study of more than 5,400 new leaders, the authors found that those who were struggling tended to exhibit five behaviors characteristic of people overly intent on securing a quick win. They focused too much on details, reacted negatively to criticism, intimidated others, jumped to conclusions, and micromanaged their direct reports. Some managed to eke out a win anyway, but the fallout was often toxic. The leaders who were thriving in their new roles, by contrast, shared not only a strong focus on results--necessary for early successes--but also excellent change-management skills. They communicated a clear vision, developed constructive relationships, and built team capabilities. They seemed to realize that the lasting value of their accomplishment would be the way they managed their teams through the transition. Collective quick wins established credibility and prepared them to lead their teams to harder-won victories. The authors provide a diagnostic tool for identifying opportunities for collective quick wins, and they share some advice for organizations: When grooming new leaders, don't just shore up their domain knowledge and technical skills; help them develop the change-management skills they will need as they settle in with their new teams.


Catecholamine response to exercise in children with attention deficit hyperactivity disorder.
The objective of this study was to examine differences in catecholamine (CA) response to exercise between children who had received a diagnosis of attention-deficit/hyperactivity disorder (ADHD) and age- and gender-matched controls. On the basis of the notion of a CA dysfunction in ADHD, we reasoned that the normal robust increase in circulating CA seen in response to exercise would be blunted in children with ADHD. To test this, we recruited 10 treatment-naïve children with newly diagnosed ADHD and 8 age-matched controls (all male) and measured CA response to an exercise test in which the work was scaled to each subject's physical capability. After exercise, epinephrine and norepinephrine increased in both control and ADHD subjects (p = 0.006 and p = 0.002, respectively), but the responses were substantially blunted in the ADHD group (p = 0.018) even though the work performed did not differ from controls. Circulating dopamine increased significantly in the control subjects (p < 0.016), but no increase was noted in the subjects with ADHD. Finally, a significant attenuation in the lactate response to exercise was found in ADHD (between groups, p < 0.005). Our data suggest that CA excretion after exercise challenges in children with ADHD is deficient. This deficiency can be detected using a minimally invasive, nonpharmacologic challenge.


An epidemiological study of the risk of cycling in the dark: the role of visual perception, conspicuity and alcohol use.
To curtail the rising numbers of cyclists seriously injured in road crashes, more insights are needed into the factors that contribute to these crashes. For instance, darkness is known to be associated with higher injury rates, but little is known about the relative influence of factors such as poor conspicuity, impaired perception and alcohol use among cyclists. To examine these factors, the present study analyzed the epidemiological crash data for three meteorological light conditions: daylight, late evening darkness and early morning darkness; for two crash types: crashes with (M-crashes) and without motorized traffic (NM-crashes); and for different age groups. The relative injury rates (injury risk per distance travelled in darkness corrected for daylight injury risks for each age group) confirmed findings from earlier studies that cycling in late evening darkness is associated with higher injury rates than cycling in daylight conditions. This is the case for both crash types with only small differences between the age groups suggesting that poor conspicuity (M-crashes) and impaired perception (NM-crashes) may play a role. In comparison to late evening darkness, relative injury rates in early morning darkness are much higher. This is the case for both crash types with large differences among the age groups, suggesting that in addition to the absence of daylight also age related risk factors are at play. Support for this hypothesis was found from the analyses of hospital records, showing that the proportion of seriously injured cyclists who have been drinking is highest in early morning darkness and has strongly increased over the last decades. These insights provide input for the selection of countermeasures such as improved lighting (both street and bicycle lights) and interventions targeting alcohol use among cyclists.


Effects of cessation of a policy of neonatal fluconazole prophylaxis on fungal resurgence.
Fluconazole has been used as prophylaxis against systemic fungal infections in preterm neonates. We conducted a study to determine whether cessation of a policy of prophylactic fluconazole results in a resurgence of fungal infections in a unit. Neonates born in the 3 epoches: A 36-month pre-Fluconazole prophylaxis epoch (Group 1), a 21-month Fluconazole prophylaxis epoch (Group 2) and a 39-month post Fluconazole prophylaxis epoch (Group 3) were compared for incidence and onset of fungal sepsis and resistance patterns. There was a decline in the incidence of fungal sepsis from Group 1 to Group 2, and it remained stable from Group 2 to Group 3. There was no significant difference in resistance to Fluconazole and to any of the azoles in Groups 1, 2 and 3 respectively.


MR imaging of newborns by using an MR-compatible incubator with integrated radiofrequency coils: initial experience.
To meet the needs of term and preterm neonates undergoing magnetic resonance (MR) imaging, an MR-compatible incubator with air, temperature, and humidity regulators and integrated radiofrequency coils was evaluated. Nine brain, two cardiac, and two pelvic examinations were performed by using a 1.5-T clinical MR imaging unit. The axillary temperature of the newborns varied by less than 0.8 degrees C, their vital signs remained stable, and no complications were encountered. The diagnostic quality of images obtained with the MR-compatible incubator was superior to that of images obtained with the standard MR imaging equipment. The use of an MR-compatible incubator for examinations of ill neonates is feasible and safe and yields excellent MR images.


Text messaging promoting physician-led brief intervention tobacco cessation: A before-and-after study among physicians in three tertiary hospitals in Nigeria.
Background: Physician-led smoking cessation services are suboptimal in Nigeria. Objectives: This study evaluated a text messaging intervention designed to increase the knowledge and practices of physicians in Nigeria to help smokers quit. Methods: Using a pre-post study design, all physicians (N = 946) in 3 tertiary care hospitals located in 3 geopolitical zones in Nigeria were sent 2-3 text messages weekly over a 13-week period to create awareness and improve cessation practices using the "Ask, Advise and Refer" (AAR) model. The primary outcomes were the awareness of AAR and the proportion of physicians who offered each of the components of the brief intervention (AAR) to at least half of eligible patients. Secondary outcomes included the attitudes and self-reported effects of the messages on motivation to offer AAR to patients who smoke. Results: Of the 946 eligible respondents, only 165 responded to both the before and after intervention surveys (17.4% participation rate). Participants were more likely to indicate awareness of the AAR approach after the intervention (60%) than before (21.2%). Overall, physicians' practice of each component of the AAR changed significantly after the intervention (P < .001; McNemar test). Of the participants, 71.5% reported reading the messages most/all of the time and 84.8% reported that the frequency of the messages was just adequate. Conclusions: A brief and low-cost text messaging intervention to physicians increased the awareness and practice of AAR in those who participated in the study. However, the relatively low participation rate highlights the importance of new research to improve and expand text messaging as an intervention among physicians to help them foster tobacco treatment among their patients.


Cognitive and psychomotor performance during alcohol hangover.
The consequences of alcohol consumption have risen high on health and social agendas in recent years. Although much work has focused on the physical problems associated with alcohol use, one theme that has emerged in alcohol research has been a focus on the effects of hangovers on functioning. This brief literature review specifically examines recent empirical investigations of the relationship between alcohol hangover and psychological performance and is tabled as an update to our earlier review of similar research (Stephens et al., 2008). A literature search generated 75 results on hangover and cognition (and synonyms) since the last published review. However, of these, only 4 met all inclusion criteria, such as establishing that BAL (Blood Alcohol Level) was zero at testing. Taking the findings of these newer studies with those that we reviewed previously, there appears to be real evidence of convergence of findings. There are now four rigorous laboratory studies, two less rigorous laboratory studies lacking placebo control and two rigorous naturalistic studies that indicate specific cognitive decrements in attention and memory during the hangover phase of alcohol consumption. Given this convergence, research agendas for increasing understanding of the cognitive effects of alcohol hangover should now switch from studies that routinely assess many cognitive functions to studies assessing the attention and memory deficits of hangover in greater detail.


Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa.
Defective sperm function is the largest defined cause of human infertility; however, the etiology of this condition is poorly understood. Although oxidative stress is acknowledged as a key contributor to this pathology, there are also data indicating that defective human spermatozoa contain abnormally high amounts of cis-unsaturated fatty acids. This study investigated whether a causative relationship exists between these two attributes of impaired semen quality. The objective of this study was to determine whether polyunsaturated fatty acids can induce oxidative stress in human spermatozoa. Dihydroethidium and SYTOX Green were used in conjunction with flow cytometry and HPLC to investigate reactive oxygen species (ROS) generation by human spermatozoa after fatty acid exposure. Arachidonic acid (AA) induced a time- and dose-dependent increase in ROS generation by human spermatozoa that led to the promotion of peroxidative damage and a loss of sperm motility. This effect could not be blocked with inhibitors of the cyclooxygenase or lipoxygenase pathways of AA metabolism, rotenone, protein kinase C antagonists, or known inhibitors of plasma membrane redox systems. However, ROS generation could be triggered with other cis-unsaturated fatty acids including linoleic and docosahexaenoic acids. Saturated fatty acids, methyl esters of unsaturated fatty acids, or other amphiphiles were all ineffective. However in a cell-free system, AA could trigger a redox signal via mechanisms that were profoundly disrupted by diphenylene iodonium, a flavoprotein inhibitor. The presence of excess unsaturated fatty acids in defective human spermatozoa may precipitate the oxidative stress encountered in male infertility.


Genetic history of an archaic hominin group from Denisova Cave in Siberia.
Using DNA extracted from a finger bone found in Denisova Cave in southern Siberia, we have sequenced the genome of an archaic hominin to about 1.9-fold coverage. This individual is from a group that shares a common origin with Neanderthals. This population was not involved in the putative gene flow from Neanderthals into Eurasians; however, the data suggest that it contributed 4-6% of its genetic material to the genomes of present-day Melanesians. We designate this hominin population 'Denisovans' and suggest that it may have been widespread in Asia during the Late Pleistocene epoch. A tooth found in Denisova Cave carries a mitochondrial genome highly similar to that of the finger bone. This tooth shares no derived morphological features with Neanderthals or modern humans, further indicating that Denisovans have an evolutionary history distinct from Neanderthals and modern humans.


Modernizing methodology for the WHO assessment of substances for the international drug control conventions.
The WHO Executive Board revised the guidance that governs the procedures for the WHO review of psychoactive substances for international drug control in 2010. To meet the standards defined in these guidelines, the current evaluation methodology at WHO must be an evidence-based assessment. We describe the history of substance evaluation from 1912 to the present and the development of the evaluation methods over time including a description of the current assessment system, using reports from WHO and its predecessor, the League of Nations. Furthermore, we describe the current review system. We found that some substances under international control were never reviewed; other substances were reviewed decades ago. We argue that assessments do not have unlimited validity, and therefore, substances need to be re-assessed periodically, as already recommended by the Expert Committee on Drug Dependence in 1982. We propose that the evaluation time be shortened; that the influence of the route of administration and/or dosage form of the preparation is considered in the evaluation; and we recommend studying national and regional assessment systems and adopting their best practices. With this article, we make a case for the inclusion of systematic review and other methods of comprehensive analysis of substance evaluation to arrive at a process of equal rigour and quality as already applied by WHO for the development of treatment guidelines.


Target-specific short-term dynamics are important for the function of synapses in an oscillatory neural network.
Short-term dynamics such as facilitation and depression are present in most synapses and are often target-specific even for synapses from the same type of neuron. We examine the dynamics and possible functions of two synapses from the same presynaptic neuron in the rhythmically active pyloric network of the spiny lobster. Using simultaneous recordings, we show that the synapses from the lateral pyloric (LP) neuron to the pyloric dilator (PD; a member of the pyloric pacemaker ensemble) and the pyloric constrictor (PY) neurons both show short-term depression. However, the postsynaptic potentials produced by the LP-to-PD synapse are larger in amplitude, depress less, and recover faster than those produced by the LP-to-PY synapse. The main function of the LP-to-PD synapse is to slow down the pyloric rhythm. However, in some cases, it slows down the rhythm only when it is fast and has no effect or to speeds up when it is slow. In contrast, the LP-to-PY synapse functions to delay the activity of the PY neuron; this delay increases as the cycle period becomes longer. Using a computational model, we show that the short-term dynamics of synaptic depression observed for each of these synapses are tailored to their individual functions and that replacing the dynamics of either synapse with the other would disrupt these functions. Together, the experimental and modeling results suggest that the target-specific features of short-term synaptic depression are functionally important for synapses efferent from the same presynaptic neuron.


Regulation of mouse Heparanase gene expression in T lymphocytes and tumor cells.
Heparanase (HPSE) is an endoglycosidase that cleaves heparan sulfate (HS) and plays an important role in tumor metastasis, angiogenesis and inflammation. The regulation of HPSE expression and function is tightly controlled and the increasing use of the mouse as an animal model to define the role of HPSE in many physiological and pathological settings, makes understanding the regulatory mechanisms of HPSE in this species of fundamental importance. However, the expression distribution of the mouse Hpse gene and the mechanisms that regulate its transcription are poorly defined. In this study, the mouse Hpse gene was determined to encode for two mRNA transcripts of 1.9 and 3.2 kb in length with identical open reading frames that showed similar tissue expression distribution to the human HPSE. The mouse Hpse promoter was cloned and a 478-bp minimal promoter was identified that contained regulatory elements responsible for both basal promoter activity in mouse tumor cells as well as inducible activity in T cells. Mutagenesis and transactivation studies identified a functional site in the minimal promoter region for the transcription factor Early growth response gene 1 (Egr1). Interestingly, Egr1 acted differentially in mouse tumor cells, functioning in an activating or repressive manner in breast carcinoma or melanoma cells, respectively. Furthermore, the proximal region of the promoter, identified as important in the regulation of Hpse transcription, was shown to become accessible in T cells upon cell activation. Significantly, the maximal accessibility of the promoter occurred at 16 h post-stimulation, which correlated with the induction kinetics of Hpse mRNA expression. In summary, this study demonstrates that mouse Hpse is expressed and regulated in a similar manner to human HPSE and also provides some novel insights into mechanisms of Hpse gene regulation that are likely to be relevant to control of the human gene.


A prospective observational study of on-treatment plasma homocysteine levels as a biomarker of toxicity, depression and vitamin supplementation lead-in time pre pemetrexed, in patients with non-small cell lung cancer and malignant mesothelioma.
Vitamin supplementation reduces pemetrexed toxicity. Raised plasma homocysteine reflects deficiency in vitamin B12 and folate, and is suppressed by supplementation. This observational study of 112 patients receiving pemetrexed-based chemotherapy assessed homocysteine levels after 3 weeks of vitamin supplementation, hypothesising high levels would correlate with ongoing deficiency, thus increased toxicity. Primary endpoint was the composite of proportion of patients with treatment delay/ dose reduction/ drug change or hospitalisation during the first six weeks of chemotherapy, comparing those with normal plasma homocysteine (successfully supplemented, SS) and those with high homocysteine (unsuccessfully supplemented, USS). Secondary endpoints included toxicity and analyses for depression. Post-hoc analysis examined correlation between interval of vitamin and folate supplementation and pemetrexed on primary endpoint and grade 3-4 toxicities. Eighty-four patients (84%) were successfully supplemented (SS group). The proportion of patients undergoing a treatment delay/ dose reduction/ drug change or hospitalisation in SS group was 44.0% (95% confidence interval [CI] 33.2%-55.3%) and in USS group was 18.8% (95% CI 4.0%-45.6%) (p = 0.09). Twelve percent of patients gave a past history of depression however 66% of patients had an on study Hospital Anxiety and Depression (HAD) score of >7. Supplementation status was not associated with depression. The median overall survival (OS) was 11.8 months (95% CI 8.6-16.5) in the SS group and 8.8 months (95% CI 6.6-16.2) in the US group (p = 0.5). The number of days (<7 or ≥ 7 days) between vitamin B12 and folate initiation and pemetrexed administration, had no effect on the primary endpoint and grade 3-4 toxicities. On-treatment homocysteine levels were not a biomarker of toxicity or depression. Standard vitamin supplementation is adequate in the majority of patients receiving pemetrexed. High HAD score were noted in this population giving an opportunity for mental health intervention. The lead-in time for vitamin supplementation can be short.


Physical Activity and Function in Assisted Living Residents.
The purpose of this study was to consider the feasibility, reliability, and validity of MotionWatch 8. A total of 249 residents were recruited from 26 assisted living settings. Data collection included demographics, comorbidities, function (Barthel Index), physical activity (MotionWatch 8), and falls. The mean age of participants was 86.86 ( SD = 7.0), the majority were women 179 (74%) and White ( N = 232, 96%). A total of 86% of participants wore the MotionWatch 8. There were no significant differences in physical activity over 3 days of testing. The MotionWatch 8 findings were significantly associated with activities of daily living (ADL) function. There were no significant differences in ADL function or physical activity between those who did and did not fall. The study provides additional support for the feasibility, reliability, and validity of the MotionWatch 8 and confirms that older adults living in assisted living settings spend the majority of their time in sedentary activity.


Pyometra in captive large felids: a review of eleven cases.
Eleven cases of pyometra were diagnosed in a captive exotic felid collection over 3 yr in seven African lions (Panthera leo), two tigers (P. tigris), one liger (lion-tiger crossbreed), and one leopard (P. pardus). Clinical signs included anorexia, lethargy, vulvar discharge, and vomiting. Diagnosis was based on clinical signs, complete blood cell counts, plasma biochemistry and electrolyte values, radiographs, and abdominal ultrasonography. The most common findings on complete blood count and biochemistry profiles were leukocytosis (>15,000/microL) and hyperproteinemia (>8.2 g/dL) due to increased globulins. Abdominal radiographic findings were largely nonspecific, but ultrasonography routinely showed a distended, fluid-filled uterus. Each case was treated with ovariohysterectomy and systemic antibiotic therapy. Lions were shown to be at an increased risk for developing pyometra compared with other species. Pyometra should be considered as a differential diagnosis in anorexic or lethargic intact female large felids, and ovariohysterectomy may be warranted in nonbreeding female lions.


Ocean warming and tropical invaders erode the performance of a key herbivore.
Climate change and bioinvasions are two facets of global change that can act in tandem to impact native species and ecosystems. However, their combined effects on key species have rarely been studied. The Mediterranean Sea is a hot spot of both ocean warming and bioinvasions, where their impact can be tested together. In recent years, the population of a key herbivore, the European purple sea urchin (Paracentrotus lividus), has virtually collapsed along the Israeli Mediterranean coast (southeastern Levant). Here, we used field and lab experiments to test two complementary hypotheses that may explain the urchin population collapse: (1) resource competition that may lead to competitive exclusion by invasive grazers (two Red Sea rabbitfishes) and (2) reduced performance due to ocean warming. An inclusion-exclusion in situ caging experiment revealed a strong negative impact of fish grazing on algal cover and on the urchin's gut content and gonado-somatic index (GSI). Laboratory experiments revealed a considerable negative impact of both elevated temperature and food deficiency on sea urchin respiration and GSI, and consequently on its energy budget and reproductive potential and, potentially, fitness. Such reduced reproductive capacity must have greatly lowered the sea urchin's population viability, contributing (and possibly even leading) to its collapse in the southeastern Levant in the past two decades. Urchin population declines are expected to spread to the west and north of the Mediterranean Sea following further warming and rabbitfish expansion. This study is the first to demonstrate the potential additive effects of ocean warming and implied competitive exclusion by an invader on a native species at its warm biogeographic distribution edge.


Neurodevelopmental and growth outcomes after invasive Group B Streptococcus in early infancy: A multi-country matched cohort study in South Africa, Mozambique, India, Kenya, and Argentina.
Data are limited regarding long-term consequences of invasive GBS (iGBS) disease in early infancy, especially from low- and middle-income countries (LMIC) where most cases occur. We aimed to estimate risk of neurodevelopmental impairment (NDI) in children with a history of iGBS disease. A multi-country matched cohort study was undertaken in South Africa, India, Mozambique, Kenya, and Argentina from October 2019 to April 2021. The exposure of interest was defined as a history of iGBS disease (sepsis or meningitis) before 90 days of age, amongst children now aged 1·5-18 years. Age and sex-matched, children without history of GBS were also recruited. Age-appropriate, culturally-adapted assessments were used to define NDI across multiple domains (cognitive, motor, hearing, vision, emotional-behaviour, growth). Pooled NDI risk was meta-analysed across sites. Association of iGBS exposure and NDI outcome was estimated using modified Poisson regression with robust variance estimator. Amongst 138 iGBS survivors and 390 non-iGBS children, 38·1% (95% confidence interval [CI]: 30·0% - 46·6%) of iGBS children had any NDI, compared to 21·7% (95% CI: 17·7% - 26·0%) of non- iGBS children, with notable between-site heterogeneity. Risk of moderate/severe NDI was 15·0% (95% CI: 3·4% - 30·8%) among GBS-meningitis, 5·6% (95% CI: 1·5% - 13·7%) for GBS-sepsis survivors. The adjusted risk ratio (aRR) for moderate/severe NDI among iGBS survivors was 1.27 (95% CI: 0.65, 2.45), when compared to non-GBS children. Mild impairment was more frequent in iGBS (27.6% (95% CI: 20.3 - 35.5%)) compared to non-GBS children (12.9% (95% CI: 9.7% - 16.4%)). The risk of emotional-behavioural problems was similar irrespective of iGBS exposure (aRR=0.98 (95% CI: 0.55, 1.77)). Our findings suggest that iGBS disease is on average associated with a higher risk of moderate/severe NDI, however substantial variation in risk was observed between sites and data are consistent with a wide range of values. Our study underlines the importance of long-term follow-up for at-risk neonates and more feasible, standardised assessments to facilitate diagnosis in research and clinical practice. This work was supported by a grant (INV-009018) from the Bill & Melinda Gates Foundation to the London School of Hygiene &Tropical Medicine.


Patients and Medical Staff Attitudes Toward the Future Inclusion of eHealth in Tuberculosis Management: Perspectives From Six Countries Evaluated using a Qualitative Framework.
Digitally delivering healthcare services is very attractive for tuberculosis (TB) management as this disease has a complex diagnosis and lengthy management and involves multiple medical and nonmedical specialists. Especially in low- and middle-income countries, eHealth could potentially offer cost-effective solutions to bridge financial, social, time, and distance challenges. The goal of the research is to understand what would make eHealth globally applicable and gain insight into different TB situations, opportunities, and challenges. We performed focus group interviews with TB experts and patients from 6 different countries on 4 different continents. The focus group interviews followed the theory of planned behavior framework to offer structured recommendations for a versatile eHealth solution. The focus group interviews were preceded by a general demographic and technology use questionnaire. Questionnaire results were analyzed using basic statistics in Excel (Microsoft Corporation). Focus group interview data were analyzed using ATLAS.ti 8 (ATLAS.ti Scientific Software Development GmbH) by assigning codes to quotations and grouping codes into the 5 domains within the framework. A total of 29 patients and 32 medical staff members were included in our study. All medical staff had used the internet, whereas 31% (9/61) of patients had never been online. The codes with the most quotations were information in relation to eHealth (144 quotations) and communication (67 quotations). The consensus among all participants from all countries is that there are important communication and information gaps that could be bridged by an eHealth app. Participants from different countries also highlighted different challenges, such as a majority of asylum-seeker patients or lack of infrastructure that could be addressed with an eHealth app. Within the 6 countries interviewed, there is high enthusiasm toward eHealth in TB. A potential app could first target information and communication gaps in TB, with additional modules aimed at setting-specific challenges.


Associations Between Nurse-Patient Interaction and Sense of Coherence Among Cognitively Intact Nursing Home Residents.
Aim: To investigate the association between nurse-patient interaction and sense of coherence among cognitively intact nursing home residents. Method: In a cross-sectional design, data were collected in 2017 and 2018 using the Nurse-Patient Interaction Scale (NPIS) and the 13-item Sense of Coherence Scale (SOC-13). Of the 204 cognitively intact nursing home residents who met the inclusion criteria, 188 (92%) participated, representing 27 nursing homes. Multiple regression in a general linear model estimated the possible effects of the 14 NPIS items on SOC-13 sum score, the possible effects of the NPIS (sum score) on SOC-13 (sum score) as well as on the subdimensions of SOC-13, comprehensibility, meaningfulness, and manageability (both without and with adjusting for sex and age). Results: Four of the 14 NPIS items revealed highly significant correlations with SOC-13 (sum score; unadjusted and adjusted for age and gender). Furthermore, the analysis adjusted for age and gender showed significant associations for NPIS (sum score) with SOC-13 (sum score), manageability, and comprehensibility. The correlation between NPIS and meaningfulness was not statistically significant. Conclusion: Nurse-patient interaction is significantly associated with SOC-13 and its subdimensions of comprehensibility and manageability but not meaningfulness. Nurse-patient interaction might be an important resource in relation to residents' sense of coherence and its subdimensions.


Targeting the Non-catalytic RVxF Site of Protein Phosphatase-1 With Small Molecules for Ebola Virus Inhibition.
Ebola virus (EBOV) is a non-segmented negative-sense RNA virus that causes a severe human disease. The ongoing EBOV outbreak in the Eastern part of Democratic Republic of the Congo has resulted to date in over 2500 confirmed cases including over 1500 deaths. Difficulties with vaccine administration indicate the necessity for development of new general drugs and therapeutic strategies against EBOV. Host Ser/Thr protein phosphatases, particularly PP1 and PP2A, facilitate EBOV transcription by dephosphorylating the EBOV VP30 protein and switching activity of the polymerase complex toward replication. Previously, we developed small molecule 1E7-03 that targeted host protein phosphatase-1 (PP1) and induces phosphorylation of EBOV VP30 protein thus shifting transcription-replication balance and inhibiting EBOV replication. Here, we developed a new EBOV inhibitor, 1E7-07, that potently inhibits EBOV replication and displays significantly improved metabolic stability when compared to previously described 1E7-03. Proteome analysis of VP30 shows that 1E7-07 increases its phosphorylation on Thr-119 and Ser-124 over 3-fold with p < 0.001, which likely contributes to EBOV inhibition. We analyzed 1E7-07 binding to PP1 using a mass spectrometry-based protein painting approach. Combined with computational docking, protein painting shows that 1E7-07 binds to several PP1 sites including the RVxF site, C-terminal groove and NIPP1-helix binding pocket. Further analysis using surface plasmon resonance and a split NanoBiT system demonstrates that 1E7-07 binds primarily to the RVxF site. Together, detailed analysis of 1E7-07 binding to PP1 and identification of the RVxF site as the main binding site opens up an opportunity for future development of PP1-targeting EBOV inhibitors.


Acclimation and Institutionalization of the Mouse Microbiota Following Transportation.
Using animal models, the gut microbiota has been shown to play a critical role in the health and disease of many organ systems. Unfortunately, animal model studies often lack reproducibility when performed at different institutions. Previous studies in our laboratory have shown that the gut microbiota of mice can vary with a number of husbandry factors leading us to speculate that differing environments may alter gut microbiota, which in turn may influence animal model phenotypes. As an extension of these studies, we hypothesized that the shipping of mice from a mouse producer to an institution will result in changes in the type, relative abundance, and functional composition of the gut microbiota. Furthermore, we hypothesized that mice will develop a microbiota unique to the institution and facility in which they are housed. To test these hypotheses, mice of two strains (C57BL/6J and BALB/cJ), two age groups (4 week and 8 week old), and originating from two types of housing (research animal facility under conventional housing and production facilities under maximum barrier housing) were obtained from The Jackson Laboratory. Fecal samples were collected the day prior to shipping, immediately upon arrival, and then on days 2, 5, 7, and weeks 2, 4, and 9 post-arrival. Following the first post-arrival fecal collection, mice were separated into 2 groups and housed at different facilities at our institution while keeping their caging, diet, and husbandry practices the same. DNA was extracted from the collected fecal pellets and 16S rRNA amplicons were sequenced in order to characterize the type and relative abundance of gut bacteria. Principal component analysis (PCA) and permutational multivariate analysis of variance (PERMANOVA) demonstrated that both the shipping and the institution and facility in which mice were housed altered the gut microbiota. Phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) predicted differences in functional composition in the gut microbiota of mice based on time of acclimation.


Drug-polymer miscibility, interactions, and precipitation inhibition studies for the development of amorphous solid dispersions for the poorly soluble anticancer drug flutamide.
The major goal of this research was to successfully formulate solid dispersion (SD) of the poorly soluble anticancer drug flutamide (FLT) using various hydrophilic polymers. Furthermore, to get more insight into SD, solid-state studies (miscibility and molecular interaction) were correlated with solution study (precipitation inhibition, dissolution). Hydrophilic polymers like PVP K90, HPMC, Eudragit EPO, and PEG 8000 were used at different drug-to-polymer w/w ratios. Solid-state miscibility studies were carried out using modulated differential scanning calorimetry (MDSC). SDs were prepared using solvent-evaporation technique and characterized by powder X-ray diffraction (PXRD) and MDSC. Infrared, Raman spectroscopy and molecular modeling were used to investigate drug-polymer interactions in the dispersions. Precipitation inhibition studies were carried out at various FLT-hydrophilic polymer ratios. Precipitation inhibition studies showed that PEG 8000 has the highest efficiency, followed by PVP K90, while HPMC and EPO showed no effect on precipitation inhibition. In the solid-state, MDSC of the physical mixture (PM) suggested that FLT is miscible to a greater extent with EPO and PEG 8000. Characterization of the amorphous dispersions using MDSC and PXRD concluded that FLT transformed from crystalline to amorphous form in the presence of PVP K90 and PEG 8000. Spectroscopic results confirmed stronger interaction of FLT with PVP K90 and PEG 8000, thereby confirming the in-solution precipitation and molecular modeling binding energy results. Amorphous dispersions formulated with PVP and PEG were stable and showed higher dissolution, an important property necessary to improve the physicochemical properties and drug delivery of poorly soluble anticancer drug FLT.


ABPS: An R Package for Calculating the Abnormal Blood Profile Score.
The Abnormal Blood Profile Score (ABPS) is used to identify blood doping in sport. It combines seven hematological markers, including hemoglobin level, reticulocytes percent, and haematocrit level, using two different machine learning algorithms in order to create a single score that has a better ability to identify doping than each parameter taken alone. The resulting score allows the detection of several types of doping using a single score and is part of the current Athlete Biological Passport program managed by World Anti-Doping Agency (WADA). We describe ≪ ABPS ≫, an R package that allows the calculation of this score. This is the first software implementation calculating this score that is released publicly. The package also contains functions to calculate the OFF-score (another score used for detection of doping), as well as several test datasets. The package is useful for laboratories conducting anti-doping analyses and for researchers working on anti-doping research projects. In particular, it has been successfully used in projects estimating the prevalence of blood doping.


Association of the human invariant chain with H-2 Db class I molecules.
We describe two proteins of 24 kDa and 33 kDa (p24 and p33) which associate with H-2 Kb and H-2 Db molecules, respectively, in human cells transfected with H-2 Kb and H-2 Db genes. This association is particularly clear in the mutant cell line T2, in which association of endogenous peptide with newly synthesized class I molecules may not occur (V. Cerundolo et al., Nature 1990. 345: 449). We show that p33 is the 33-kDa form of the human invariant chain which is resident in the endoplasmic reticulum of T2 cells (P. Cresswell, Cold Spring Harbor Symp. Quant. Biol. 1989. LIV:309). The stability of the invariant chain H-2 Db complex is critically dependent upon occupation of the class I binding site by peptide ligand. In the absence of peptide, the complex is stable at 4 degrees C whereas following exposure to peptide, the invariant chain dissociates rapidly from H-2 Db molecules (half-life of 30 min at 4 degrees C). Although the interaction between the human invariant chain and murine H-2 Db is unlikely to have any functional significance, the peptide-induced dissociation of the invariant chain is consistent with a conformational change in H-2 Db on peptide binding.


The diagnostic value of endoprobe for small esophageal leiomyomas derived from the muscularis mucosae.
Esophageal leiomyoma derived from the muscularis mucosae (MM) is a rare condition, and the optimal modality for diagnosis and treatment is controversial. Endoscopic ultrasonography can provide an accurate image of esophageal layer structure, providing information on lesion suitability for potential endoscopic therapy. We attempted to investigate the diagnostic value of a transendoscopic balloon-tipped miniature ultrasonic endoprobe for small esophageal leiomyomas derived from MM. We resected 7 small esophageal leiomyomas derived from MM by endoscopic mucosal resection (EMR), all of which were diagnosed by a balloon-tipped endoprobe. The endosonographic and pathologic features of 7 cases of small esophageal leiomyomas derived from MM were compared. The balloon-tipped endoprobe clearly showed all 7 small esophageal leiomyomas derived from MM, even those under 5 mm in size (smallest lesion, 3.0 mm). The endosonographic characteristics of small esophageal leiomyomas derived from MM were a hypoechoic mass with smooth, regular, and a well-defined outer margin and homogenous inner echogram arising from the second hypoechoic layer. Complete resections were possible in all 7 cases by EMR without any complications. Tumor size was 3.0-13.5 mm (mean 7.8 mm) in maximum diameter. In all cases, endosonographic findings by endoprobe were exactly concordant with pathologic finding in determining the tumors depth in the esophageal wall, tissue origin and characteristics, growth pattern, and size. We detail the balloon-tipped endoprobe is a simple, convenient, and very useful in making accurate diagnosis of small esophageal leiomyomas derived from the MM and the appropriate applications of EMR.


Glu-Q-tRNA(Asp) synthetase coded by the yadB gene, a new paralog of aminoacyl-tRNA synthetase that glutamylates tRNA(Asp) anticodon.
Analysis of the completed genome sequences revealed presence in various bacteria of an open reading frame (ORF) encoding a polypeptide chain presenting important similarities with the catalytic domain of glutamyl-tRNA synthetases but deprived of the C-terminal anticodon-binding domain. This paralog of glutamyl-tRNA synthetases, the YadB protein, activates glutamate in the absence of tRNA and transfers the activated glutamate not on tRNA(Glu) but instead on tRNA(Asp). It has been shown that tRNA(Asp) is able to accept two amino acids: aspartate charged by aspartyl-tRNA synthetase and glutamate charged by YadB. The functional properties of YadB contrast with those of the canonical glutamyl-tRNA synthetases, which activate Glu only in presence of the cognate tRNA before aminoacylation of the 3'-end of tRNA. Biochemical approaches and mass spectrometry investigations revealed that YadB transfers the activated glutamate on the cyclopenthene-diol ring of the modified nucleoside queuosine posttranscriptionally inserted at the wobble position of the anticodon-loop to form glutamyl-queuosine. Unstability of the ester bond between the glutamate residue and the cyclopenthene-diol (half-life 7.5 min) explains why until now this modification escaped detection. Among Escherichia coli tRNAs containing queuosine in the wobble position, only tRNA(Asp) is substrate of YadB. Sequence comparison reveals a structural mimicry between the anticodon-stem and loop of tRNA(Asp) and the amino acid acceptor-stem of tRNA(Glu). YadB, renamed glutamyl-Q-tRNA(Asp) synthetase, constitutes the first enzyme structurally related to aminoacyl-tRNA synthetases which catalyzes a hypermodification in tRNA, and whose function seems to be conserved among prokaryotes. The discovery of glutamyl-Q-tRNA(Asp) synthetase breaks down the current paradigm according to which the catalytic domain of aminoacyl-tRNA synthetases recognizes the amino acid acceptor-stem of tRNA and aminoacylates the 3'-terminal ribose. The evolutionary significance of the existence of an aminoacyl-tRNA synthetase paralog dedicated to the hypermodification of a tRNA anticodon will be discussed.


Photodynamic inactivation of conidia of the fungi Metarhizium anisopliae and Aspergillus nidulans with methylene blue and toluidine blue.
Antimicrobial photodynamic treatment (PDT) is a promising method that can be used to control localized mycoses or kill fungi in the environment. A major objective of the current study was to compare the conidial photosensitization of two fungal species (Metarhizium anisopliae and Aspergillus nidulans) with methylene blue (MB) and toluidine blue (TBO) under different incubation and light conditions. Parameters examined were media, photosensitizer (PS) concentration and light source. PDT with MB and TBO resulted in an incomplete inactivation of the conidia of both fungal species. Conidial inactivation reached up to 99.7%, but none of the treatments was sufficient to achieve a 100% fungicidal effect using either MB or TBO. PDT delayed the germination of the surviving conidia. Washing the conidia to remove unbound PS before light exposure drastically reduced the photosensitization of A. nidulans. The reduction was much smaller in M. anisopliae conidia, indicating that the conidia of the two species interact differently with MB and TBO. Conidia of green and yellow M. anisopliae mutants were less affected by PDT than mutants with white and violet conidia. In contrast to what occurred in PBS, photosensitization of M. anisopliae and A. nidulans conidia was not observed when PDT was performed in potato dextrose media.


Small molecule-gold nanorod conjugates selectively target and induce macrophage cytotoxicity towards breast cancer cells.
Gold nanoparticles are known to activate anti-tumor potential in macrophage immune cells; however, the subsequent effects of these cells on others nearby are poorly understood. A novel gold-nanoparticle conjugate that selectively targets and induces cytotoxic activity of tumor-associated macrophages towards breast cancer cells in co-culture is synthesized. These constructs are promising new tools for studying fundamental biological interactions with nanoscale materials and candidates for emerging macrophage-mediated delivery applications.


Causes and Predictors of 30-Day Readmission in Patients With Acute Myocardial Infarction and Cardiogenic Shock.
Acute myocardial infarction (AMI) occurs as a result of irreversible damage to cardiac myocytes secondary to lack of blood supply. Cardiogenic shock complicating AMI has significant associated morbidity and mortality, and data on postdischarge outcomes are limited. We derived the study cohort of patients with AMI and cardiogenic shock from the 2013 to 2014 Healthcare Cost and Utilization Project National Readmission Database. Incidence, predictors, and causes of 30-day readmissions were analyzed. From 43 212 index admissions for AMI with cardiogenic shock, 26 016 (60.2%) survived to discharge and 5277 (20.2% of survivors) patients were readmitted within 30 days. More than 50% of these readmissions occurred within first 10 days. Cardiac causes accounted for 42% of 30-day readmissions (heart failure 20.6%; acute coronary syndrome 11.6%). Among noncardiac causes, respiratory (11.4%), infectious (9.4%), medical or surgical care complications (6.3%), gastrointestinal/hepatobiliary (6.5%), and renal causes (4.8%) were most common. Length of stay ≥8 days (odds ratio [OR], 2.04; 95% confidence interval [CI], 1.70-2.44; P<0.01), acute deep venous thrombosis (OR, 1.26; 95% CI, 1.08-1.48; P<0.01), liver disease (OR, 1.25; 95% CI, 1.03-1.50; P=0.02), systemic thromboembolism (OR, 1.21; 95% CI, 1.02-1.44; P=0.02), peripheral vascular disease (OR, 1.16; 95% CI, 1.07-1.27; P<0.01), diabetes mellitus (OR, 1.16; 95% CI, 1.08-1.24; P<0.01), long-term ventricular assist device implantation (OR, 1.77; 95% CI, 1.23-2.55; P<0.01), intraaortic balloon pump use (OR, 1.10; 95% CI, 1.02-1.18; P<0.01), performance of coronary artery bypass grafting (OR, 0.85; 95% CI, 0.77-0.93; P<0.01), private insurance (OR, 0.72; 95% CI, 0.64-0.80; P<0.01), and discharge to home (OR, 0.85; 95% CI, 0.73-0.98; P=0.03) were among the independent predictors of 30-day readmission. In-hospital mortality and 30-day readmission in cardiogenic shock complicating AMI are significantly elevated. Patients are readmitted mainly for noncardiac causes. Identification of high-risk factors may guide interventions to improve outcomes within this population.


A Tool for Shared Decision Making on Referral for Prostate Biopsy in the Primary Care Setting: Integrating Risks of Cancer with Life Expectancy.
Prostate cancer (PCa) testing involves a complex individually based decision making process. It should consider competing risks from other comorbidities when estimating a survival benefit from the early detection of clinically significant (cs)PCa. We aimed to develop a prediction tool that provides concrete advice for the general practitioner (GP) on whether to refer a man for further assessment. We hereto combined the probability of detecting csPCa and the potential overall survival benefit from early detection and treatment. The PCa detection probabilities were derived from 3616 men enrolled in the Dutch arm of the European Randomized Study of Screening for Prostate Cancer (ERSPC). Survival estimates were derived from 19,834 men from the Surveillance, Epidemiology, and End Results (SEER) registry, ERSPC, and Dutch life tables. Treatment benefit was estimated from the Prostate Cancer Intervention versus Observation Trial (PIVOT, n = 731). The prediction of csPCa detection was based on prostate-specific antigen (PSA), age, %freePSA, and digital rectal examination (DRE). The life expectancy (LE) for patients with PCa receiving no treatment was adjusted for age and Charlson comorbidity index. A negative impact on LE and treatment benefit was found with higher age and more comorbidity. The proposed integrated approach may support triage at GP practices, as PCa is a heterogeneous disease in predominantly elderly men.


Comparative Structural Analysis of 20S Proteasome Ortholog Protein Complexes by Native Mass Spectrometry.
Ortholog protein complexes are responsible for equivalent functions in different organisms. However, during evolution, each organism adapts to meet its physiological needs and the environmental challenges imposed by its niche. This selection pressure leads to structural diversity in protein complexes, which are often difficult to specify, especially in the absence of high-resolution structures. Here, we describe a multilevel experimental approach based on native mass spectrometry (MS) tools for elucidating the structural preservation and variations among highly related protein complexes. The 20S proteasome, an essential protein degradation machinery, served as our model system, wherein we examined five complexes isolated from different organisms. We show that throughout evolution, from the Thermoplasma acidophilum archaeal prokaryotic complex to the eukaryotic 20S proteasomes in yeast (Saccharomyces cerevisiae) and mammals (rat - Rattus norvegicus, rabbit - Oryctolagus cuniculus and human - HEK293 cells), the proteasome increased both in size and stability. Native MS structural signatures of the rat and rabbit 20S proteasomes, which heretofore lacked high-resolution, three-dimensional structures, highly resembled that of the human complex. Using cryoelectron microscopy single-particle analysis, we were able to obtain a high-resolution structure of the rat 20S proteasome, allowing us to validate the MS-based results. Our study also revealed that the yeast complex, and not those in mammals, was the largest in size and displayed the greatest degree of kinetic stability. Moreover, we also identified a new proteoform of the PSMA7 subunit that resides within the rat and rabbit complexes, which to our knowledge have not been previously described. Altogether, our strategy enables elucidation of the unique structural properties of protein complexes that are highly similar to one another, a framework that is valid not only to ortholog protein complexes, but also for other highly related protein assemblies.


A methodology to find the elementary landscape decomposition of combinatorial optimization problems.
A small number of combinatorial optimization problems have search spaces that correspond to elementary landscapes, where the objective function f is an eigenfunction of the Laplacian that describes the neighborhood structure of the search space. Many problems are not elementary; however, the objective function of a combinatorial optimization problem can always be expressed as a superposition of multiple elementary landscapes if the underlying neighborhood used is symmetric. This paper presents theoretical results that provide the foundation for algebraic methods that can be used to decompose the objective function of an arbitrary combinatorial optimization problem into a sum of subfunctions, where each subfunction is an elementary landscape. Many steps of this process can be automated, and indeed a software tool could be developed that assists the researcher in finding a landscape decomposition. This methodology is then used to show that the subset sum problem is a superposition of two elementary landscapes, and to show that the quadratic assignment problem is a superposition of three elementary landscapes.


Incidence, diagnosis, and treatment of sinus graft infection after sinus floor elevation: a clinical study.
The aims of this clinical study were to evaluate the occurrence of sinus graft infection and the results of a planned surgical and pharmacologic treatment regimen to eliminate infections and preserve the graft. Patients were consecutively treated with sinus floor elevations using the lateral window technique. If a clinical diagnosis of sinus graft infection was made for a patient, a postoperative computed tomographic scan was performed to diagnose involvement of the sinus cavity. Of 198 patients treated with 274 sinus grafts, 8 (2.3%) developed a sinus graft infection. These patients were treated with the same surgical and pharmacologic regimen. Symptoms of infection were recognized, the infected graft was removed, and the remaining graft material was cleansed. The remaining defect was not filled with new graft material; instead, it was left to heal for an extended healing period (average of 10.6 months), and residual bone defects were corrected during implant placement. Acute symptoms of infection disappeared within 48 hours of treatment and all patients healed uneventfully thereafter. All 24 implants placed have survived to date (with implant loading times ranging from 1 to 9 years), resulting in a 100% implant survival rate for these sites with postoperative infection. The overall clinical outcome, including patient satisfaction, achievement of optimal graft volume without subsequent sinus elevation, and long-term implant survival, confirmed the success of this treatment protocol.


Biomolecular characterization of the levansucrase of Erwinia amylovora, a promising biocatalyst for the synthesis of fructooligosaccharides.
Erwinia amylovora is a plant pathogen that affects Rosaceae, such as apple and pear. In E. amylovora the fructans, produced by the action of a levansucrase (EaLsc), play a role in virulence and biofilm formation. Fructans are bioactive compounds, displaying health-promoting properties in their own right. Their use as food and feed supplements is increasing. In this study, we investigated the biomolecular properties of EaLsc using HPAEC-PAD, MALDI-TOF MS, and spectrophotometric assays. The enzyme, which was heterologously expressed in Escherichia coli in high yield, was shown to produce mainly fructooligosaccharides (FOSs) with a degree of polymerization between 3 and 6. The kinetic properties of EaLsc were similar to those of other phylogenetically related Gram-negative bacteria, but the good yield of FOSs, the product spectrum, and the straightforward production of the enzyme suggest that EaLsc is an interesting biocatalyst for future studies aimed at producing tailor-made fructans.


Robust induction of TRMs by combinatorial nanoshells confers cross-strain sterilizing immunity against lethal influenza viruses.
Antigen-specific lung-resident memory T cells (TRMs) constitute the first line of defense that mediates rapid protection against respiratory pathogens and inspires novel vaccine designs against infectious pandemic threats, yet effective means of inducing TRMs, particularly via non-viral vectors, remain challenging. Here, we demonstrate safe and potent induction of lung-resident TRMs using a biodegradable polymeric nanoshell that co-encapsulates antigenic peptides and TLR9 agonist CpG-oligodeoxynucleotide (CpG-ODN) in a virus-mimicking structure. Through subcutaneous priming and intranasal boosting, the combinatorial nanoshell vaccine elicits prominent lung-resident CD4+ and CD8+ T cells that surprisingly show better durability than live viral infections. In particular, nanoshells containing CpG-ODN and a pair of conserved class I and II major histocompatibility complex-restricted influenza nucleoprotein-derived antigenic peptides are demonstrated to induce near-sterilizing immunity against lethal infections with influenza A viruses of different strains and subtypes in mice, resulting in rapid elimination of replicating viruses. We further examine the pulmonary transport dynamic and optimal composition of the nanoshell vaccine conducive for robust TRM induction as well as the benefit of subcutaneous priming on TRM replenishment. The study presents a practical vaccination strategy for inducing protective TRM-mediated immunity, offering a compelling platform and critical insights in the ongoing quest toward a broadly protective vaccine against universal influenza as well as other respiratory pathogens.


Detecting DNA depurination with solid-state nanopores.
Among the different types of DNA damage that occur endogenously in the cell, depurination is especially prevalent. These lesions can initiate mutagenesis and have been implicated in a variety of diseases, including cancer. Here, we demonstrate a new approach for the detection of depurination at the single-molecule scale using solid-state nanopores. We induce depurination in short duplex DNA using acidic conditions and observe that the presence of apurinic sites results in significantly slower dynamics during electrokinetic translocation. This procedure may be valuable as a diagnostic for in situ quantification of DNA depurination.


Identification of testicular cancer driver genes by a cross-species comparative oncology approach.
Germ cell tumors arise in the testis, ovary, or extragonadal locations and have a wide range of histopathological and clinical presentations. The relative lack of animal models of germ cell tumors has impeded functional assessment of candidate driver genes. Previously, we described the development of testicular germ cell tumors in zebrafish carrying a mutation in bmpr1bb, a BMP family receptor, and demonstrated that human germ cell tumors have defects in BMP signaling. To further credential the zebrafish model for studies of human germ cell tumor, and to elucidate conserved genetic programs underlying the development of germ cell tumor. We used genetic techniques to ablate the germ cell lineage in developing fish and tested tumors for loss-of-heterozygosity of the wild-type allele of bmpr1bb. We performed comparative gene expression profiling of zebrafish and human germ cell tumors and carried out functional studies of selected genes. Ablation of germ cells completely prevents testis tumor formation in the fish, definitively establishing the germ cell origin of the tumors. Germ cell tumors in bmpr1bb heterozygous mutants retain the wild-type allele, indicating haploinsufficiency of bmpr1bb as the mechanism of tumor formation. Comparison of RNA-Seq and microarray data from human and zebrafish germ cell tumors revealed a unique overlapping signature shared by the zebrafish tumors with human seminomas, yolk sac tumors, and embryonal carcinomas. The most highly conserved gene set in this cross-species analysis included potential driver genes such as JUP, which we show to be essential for germ cell tumor cell growth. Our findings highlight the value of cross-species comparative oncology for the identification of candidate human cancer genes.


Audiovisual synchrony perception for music, speech, and object actions.
We investigated the perception of synchrony for complex audiovisual events. In Experiment 1, a series of music (guitar and piano), speech (sentences), and object action video clips were presented at a range of stimulus onset asynchronies (SOAs) using the method of constant stimuli. Participants made unspeeded temporal order judgments (TOJs) regarding which stream (auditory or visual) appeared to have been presented first. Temporal discrimination accuracy was significantly better for the object actions than for the speech video clips, and both were significantly better than for the music video clips. In order to investigate whether or not these differences in TOJ performance were driven by differences in stimulus familiarity, we conducted a second experiment using brief speech (syllables), music (guitar), and object action video clips of fixed duration together with temporally reversed (i.e., less familiar) versions of the same stimuli. The results showed no main effect of stimulus type on temporal discrimination accuracy. Interestingly, however, reversing the video clips resulted in a significant decrement in temporal discrimination accuracy as compared to the normally presented for the music and object actions clips, but not for the speech stimuli. Overall, our results suggest that cross-modal temporal discrimination performance is better for audiovisual stimuli of lower complexity as compared to stimuli having continuously varying properties (e.g., syllables versus words and/or sentences).


Development of a tool to evaluate health science students' experiences of an interprofessional education (IPE) programme.
The Rural Interprofessional Programme Emergency Retreat (RIPPER) is an educational programme collaboratively developed and evaluated by an interprofessional team from Schools within Faculty of Health Science (FHS), University of Tasmania (UTAS), Australia. The aims of RIPPER are to foster and facilitate positive and productive interprofessional learning experiences for undergraduate students in a rural setting; and to develop a firmly embedded and sustainable interprofessional healthcare module within the health science curriculum. This paper reports on the development of a reliable and valid survey tool to evaluate students' understandings and experiences of this interprofessional learning programme. Twenty-nine students from the Schools of Nursing, Medicine and Pharmacy of the FHS, UTAS participated in the RIPPER programme which offers a number of interactive rural emergency healthcare scenarios using high- and low-fidelity simulation. To evaluate the programme a survey which consisted of 2 main components was developed and implemented before and after the programme. The first component was designed to gather students' demographic information, their understanding of the interprofessional practice concepts, and their expectations of the RIPPER programme using open-ended questions. The second component consisted of a 5-point Likert scale for students to rank their level of agreement pre- and post- intervention with 12 statements about team working, programme evaluation and collaborative learning. Three processes were used to establish the validity and reliability of the survey. Content validity was assessed by academics and experts in health science education. Construct validity was assessed using exploratory factor analysis. The internal consistency and reliability of the survey was checked using Cronbach's alpha coefficient. Factor analysis of the 12 statements identified 3 main factors including appreciation of professional roles and responsibilities, improved professional practice based on effective teamwork and the importance of students learning and working together for improved clinical practice. Reliability of the survey was established. The survey is able to evaluate students' understandings and experiences of this interprofessional learning programme.


The uptake, retention and clearance of drug-loaded dendrimer nanoparticles in astrocytes - electrophysiological quantification.
Nanoparticle-based drug delivery systems may impose risks to patients due to potential toxicity associated with a lack of clearance from cells or prolonged carrier-cell retention. This work evaluates vesicular cell uptake, retention and the possible transfer of endocytosed methylprednisolone-loaded carboxymethylchitosan/poly(amidoamine) dendrimer nanoparticles (NPs) into secretory vesicles of rat cultured astrocytes. The cells were incubated with NPs and unitary vesicle fusions/fissions with the plasma membrane were monitored employing high-resolution membrane capacitance measurements. In the NP-treated cells the frequency of unitary exocytotic events was significantly increased. The presence of NPs also induces an increase in the size of exocytotic vesicles interacting with the plasma membrane, which exhibit transient fusion with prolonged fusion pore dwell-time. Live-cell confocal imaging revealed that once NPs internalize into endocytotic compartments they remain in the cell for 7 days, although a significant proportion of these merge with secretory vesicles destined for exocytosis. Co-localization studies show the route of clearance of NPs from cells via the exocytotic pathway. These findings bring new insight into the understanding of the intracellular trafficking and biological interactions of drug-loaded dendrimer NPs targeting astrocytes.


Control of pulmonary vascular tone during exercise in health and pulmonary hypertension.
Despite the importance of the pulmonary circulation as a determinant of exercise capacity in health and disease, studies into the regulation of pulmonary vascular tone in the healthy lung during exercise are scarce. This review describes the current knowledge of the role of various endogenous vasoactive mechanisms in the control of pulmonary vascular tone at rest and during exercise. Recent studies demonstrate an important role for endothelial factors (NO and endothelin) and neurohumoral factors (noradrenaline, acetylcholine). Moreover, there is evidence that natriuretic peptides, reactive oxygen species and phosphodiesterase activity can influence resting pulmonary vascular tone, but their role in the control of pulmonary vascular tone during exercise remains to be determined. K-channels are purported end-effectors in control of pulmonary vascular tone. However, K(ATP) channels do not contribute to regulation of pulmonary vascular tone, while the role of K(V) and K(Ca) channels at rest and during exercise remains to be determined. Pulmonary hypertension is associated with alterations in pulmonary vascular function and structure, resulting in blunted pulmonary vasodilatation during exercise and impaired exercise capacity. Although there is a paucity of studies pertaining to the regulation of pulmonary vascular tone during exercise in idiopathic pulmonary hypertension, the few studies that have been performed in models of pulmonary hypertension secondary to left ventricular dysfunction suggest altered control of pulmonary vascular tone during exercise. Since the increased pulmonary vascular tone during exercise limits exercise capacity, future studies are needed to investigate the vasomotor mechanisms that are responsible for the blunted exercise-induced pulmonary vasodilatation in pulmonary hypertension.


An Evaluation of a Battery of Functional and Structural Tests as Predictors of Likely Risk of Progression of Age-Related Macular Degeneration.
To evaluate the ability of visual function and structural tests to identify the likely risk of progression from early/intermediate to advanced AMD, using the Age-Related Eye Disease Study (AREDS) simplified scale as a surrogate for risk of progression. The secondary aim was to determine the relationship between disease severity grade and the observed functional and structural deficits. A total of 100 participants whose AMD status varied from early to advanced were recruited. Visual function was assessed using cone dark adaptation, 14 Hz flicker and chromatic threshold tests and retinal structure was assessed by measuring drusen volume and macular thickness. The predictive value of the tests was estimated using ordinal regression analysis. Group comparisons were assessed using analysis of covariance. Change in cone dark adaptation (cone τ) and yellow-blue (YB) chromatic sensitivity were independent predictors for AMD progression risk (cone τ, pseudo R2 = 0.35, P < 0.001; YB chromatic threshold, pseudo R2 = 0.16, P < 0.001). The only structural predictor was foveal thickness (R2 = 0.05, P = 0.047). Chromatic sensitivity and cone dark adaptation were also the best functional tests at distinguishing between severity groups. Drusen characteristics clearly differentiated between participants with early and advanced disease, but were not able to differentiate between those with early AMD and controls. Mean differences in retinal thickness existed between severity groups at the foveal (P = 0.040) and inner (P = 0.001) subfields. This study indicates that cone τ, YB chromatic threshold and foveal thickness are independent predictors of likely risk of AMD progression.


Structural features in the Rous sarcoma virus RNA stability element are necessary for sensing the correct termination codon.
Nonsense-mediated mRNA decay (NMD) is an mRNA quality control mechanism that selectively recognizes and targets for degradation mRNAs containing premature termination codons. Retroviral full-length RNA is presented to the host translation machinery with characteristics rarely observed among host cell mRNAs: a long 3' UTR, retained introns, and multiple open reading frames. As a result, the viral RNA is predicted to be recognized by the host NMD machinery and degraded. In the case of the Rous sarcoma virus (RSV), we identified a stability element (RSE), which resides immediately downstream of the gag termination codon and facilitates NMD evasion. We defined key RNA features of the RSE through directed mutagenesis of the virus. These data suggest that the minimal RSE is 155 nucleotides (nts) and functions independently of the nucleotide sequence of the stop codon or the first nucleotide following the stop codon. Further data suggested that the 3'UTRs of the RSV pol and src may also function as stability elements. We propose that these stability elements in RSV may be acting as NMD insulators to mask the preceding stop codon from the NMD machinery.


Improving Patch-Based Convolutional Neural Networks for MRI Brain Tumor Segmentation by Leveraging Location Information.
The manual brain tumor annotation process is time consuming and resource consuming, therefore, an automated and accurate brain tumor segmentation tool is greatly in demand. In this paper, we introduce a novel method to integrate location information with the state-of-the-art patch-based neural networks for brain tumor segmentation. This is motivated by the observation that lesions are not uniformly distributed across different brain parcellation regions and that a locality-sensitive segmentation is likely to obtain better segmentation accuracy. Toward this, we use an existing brain parcellation atlas in the Montreal Neurological Institute (MNI) space and map this atlas to the individual subject data. This mapped atlas in the subject data space is integrated with structural Magnetic Resonance (MR) imaging data, and patch-based neural networks, including 3D U-Net and DeepMedic, are trained to classify the different brain lesions. Multiple state-of-the-art neural networks are trained and integrated with XGBoost fusion in the proposed two-level ensemble method. The first level reduces the uncertainty of the same type of models with different seed initializations, and the second level leverages the advantages of different types of neural network models. The proposed location information fusion method improves the segmentation performance of state-of-the-art networks including 3D U-Net and DeepMedic. Our proposed ensemble also achieves better segmentation performance compared to the state-of-the-art networks in BraTS 2017 and rivals state-of-the-art networks in BraTS 2018. Detailed results are provided on the public multimodal brain tumor segmentation (BraTS) benchmarks.


Detection of Helicobacter pylori in dental plaque and gastric biopsy samples of Turkish patients by PCR-RFLP.
Helicobacter pylori is regarded as an important pathogen playing a key role in the pathogenesis of peptic ulcer. Different studies about the mode of transmission of the microorganism report conflicting results about dental plaque as the source of H. pylori infection. In the present study we aimed to detect the presence of H. pylori in dental plaque of Turkish patients by polymerase chain reaction (PCR) and if any to do typing by restriction fragment length polymorphism (RFLP) analysis. Fifty dyspeptic patients, to whom upper gastrointestinal endoscopy was performed, were included in the study. Dental plaques were obtained before endoscopic examination. Both dental plaque and gastric biopsy samples were amplified with Ure A and Cag A gene primers. There were no positive dental plaque samples even in the 23 patients whose gastric biopsy specimens were positive. Our findings showed that there is not a correlation between dental presentation of the microorganism and H. pylori gastritis.


Optimized method for extraction of exosomes from human primary muscle cells.
Skeletal muscle is increasingly considered an endocrine organ secreting myokines and extracellular vesicles (exosomes and microvesicles), which can affect physiological changes with an impact on different pathological conditions, including regenerative processes, aging, and myopathies. Primary human myoblasts are an essential tool to study the muscle vesicle secretome. Since their differentiation in conditioned media does not induce any signs of cell death or cell stress, artefactual effects from those processes are unlikely. However, adult human primary myoblasts senesce in long-term tissue culture, so a major technical challenge is posed by the need to avoid artefactual effects resulting from pre-senescent changes. Since these cells should be studied within a strictly controlled pre-senescent division count (<21 divisions), and yields of myoblasts per muscle biopsy are low, it is difficult or impossible to amplify sufficiently large cell numbers (some 250 × 106 myoblasts) to obtain sufficient conditioned medium for the standard ultracentrifugation approach to exosome isolation.Thus, an optimized strategy to extract and study secretory muscle vesicles is needed. In this study, conditions are optimized for the in vitro cultivation of human myoblasts, and the quality and yield of exosomes extracted using an ultracentrifugation protocol are compared with a modified polymer-based precipitation strategy combined with extra washing steps. Both vesicle extraction methods successfully enriched exosomes, as vesicles were positive for CD63, CD82, CD81, floated at identical density (1.15-1.27 g.ml-1), and exhibited similar size and cup-shape using electron microscopy and NanoSight tracking. However, the modified polymer-based precipitation was a more efficient strategy to extract exosomes, allowing their extraction in sufficient quantities to explore their content or to isolate a specific subpopulation, while requiring >30 times fewer differentiated myoblasts than what is required for the ultracentrifugation method. In addition, exosomes could still be integrated into recipient cells such as human myotubes or iPSC-derived motor neurons.Modified polymer-based precipitation combined with extra washing steps optimizes exosome yield from a lower number of differentiated myoblasts and less conditioned medium, avoiding senescence and allowing the execution of multiple experiments without exhausting the proliferative capacity of the myoblasts.


Conservation and losses of non-coding RNAs in avian genomes.
Here we present the results of a large-scale bioinformatics annotation of non-coding RNA loci in 48 avian genomes. Our approach uses probabilistic models of hand-curated families from the Rfam database to infer conserved RNA families within each avian genome. We supplement these annotations with predictions from the tRNA annotation tool, tRNAscan-SE and microRNAs from miRBase. We identify 34 lncRNA-associated loci that are conserved between birds and mammals and validate 12 of these in chicken. We report several intriguing cases where a reported mammalian lncRNA, but not its function, is conserved. We also demonstrate extensive conservation of classical ncRNAs (e.g., tRNAs) and more recently discovered ncRNAs (e.g., snoRNAs and miRNAs) in birds. Furthermore, we describe numerous "losses" of several RNA families, and attribute these to either genuine loss, divergence or missing data. In particular, we show that many of these losses are due to the challenges associated with assembling avian microchromosomes. These combined results illustrate the utility of applying homology-based methods for annotating novel vertebrate genomes.


Morphological characterization and notes on the life cycle of a newly discovered variant of Loma salmonae (Putz, Hoffman & Dunbar) from a natural infection of chinook salmon, Oncorhynchus tshawytscha (Walbaum).
Two variants of Loma salmonae occur in net-pen reared chinook salmon, Oncorhynchus tshawytscha. The typical variant (OA) has a host specificity for salmonids of the genus Oncorhynchus whereas the atypical variant (SV) has a host specificity for brook trout, Salvelinus fontinalis, and in this study, the ultrastructure of the two are compared. In fish at 8 weeks post-exposure xenomas of the SV variant have a very high proportion of mature spores compared with other developmental stages, while in xenomas of the OA variant there are fewer spores and many other developmental stages. Spores of the SV variant had up to 20 turns of their polar tube whereas those of the OA variant only had 17. Furthermore, the spores of the SV variant were significantly larger than those of the OA variant. The sporophorous vesicle for both variants appears to form around a sporogonial plasmodia, which results in many spores developing within the vesicle.


Impact of virologic breakthrough and HBIG regimen on hepatitis B recurrence after liver transplantation.
The availability of hepatitis B immune globulin (HBIG) and several oral antiviral therapies has reduced but not eliminated hepatitis B virus (HBV) recurrence. We aimed to determine the rate of HBV recurrence after orthotopic liver transplantation (OLT) in relation to virologic breakthrough pre-OLT and HBIG regimens post-OLT. Data from the NIH HBV-OLT database were analyzed. A total of 183 patients transplanted between 2001 and 2007 followed for a median of 42 months (range 1-81) post-OLT were studied. At transplant, 29% were hepatitis B e antigen (HBeAg) (+), 38.5% had HBV DNA > 5 log(10) copies/mL, 74% were receiving antiviral therapy. Twenty-five patients experienced virologic breakthrough before OLT. Post-OLT, 26%, 22%, 40% and 12% of patients received intravenous (IV) high-dose, IV low-dose, intramuscular low-dose and a finite duration of HBIG, respectively as maintenance prophylaxis. All but two patients also received antiviral therapy. Cumulative rates of HBV recurrence at 1 and 5 years were 3% and 9%, respectively. Multivariate analysis showed that listing HBeAg status and HBV DNA level at OLT were the only factors associated with HBV recurrence. In conclusion, low rates of HBV recurrence can be accomplished with all the HBIG regimens used when combined with antiviral therapy including patients with breakthrough pre-OLT as long as rescue therapy is administered pre- and post-OLT.


Neuropilins Controlling Cancer Therapy Responsiveness.
Neuropilins (NRPs) are cell surface glycoproteins, acting as co-receptors for secreted Semaphorins (SEMAs) and for members of the vascular endothelial growth factor (VEGF) family; they have been initially implicated in axon guidance and angiogenesis regulation, and more recently in cancer progression. In addition, NRPs have been shown to control many other fundamental signaling pathways, especially mediated by tyrosine kinase receptors (RTKs) of growth factors, such as HGF (hepatocyte growth factor), PDGF (platelet derived growth factor) and EGF (epidermal growth factor). This enables NRPs to control a range of pivotal mechanisms in the cancer context, from tumor cell proliferation and metastatic dissemination, to tumor angiogenesis and immune escape. Moreover, cancer treatment failures due to resistance to innovative oncogene-targeted drugs is typically associated with the activity of alternative RTK-dependent pathways; and neuropilins' capacity to control oncogenic signaling cascades supports the hypothesis that they could elicit such mechanisms in cancer cells, in order to escape cytotoxic stress and therapeutic attacks. Intriguingly, several studies have recently assayed the impact of NRPs inhibition in combination with diverse anti-cancer drugs. In this minireview, we will discuss the state-of-art about the relevance of NRPs as potential predictive biomarkers of drug response, and the rationale to target these proteins in combination with other anticancer therapies.


Structural basis for telomeric G-quadruplex targeting by naphthalene diimide ligands.
The folding of the single-stranded 3' end of the human telomere into G-quadruplex arrangements inhibits the overhang from hybridizing with the RNA template of telomerase and halts telomere maintenance in cancer cells. The ability to thermally stabilize human telomeric DNA as a four-stranded G-quadruplex structure by developing selective small molecule compounds is a therapeutic path to regulating telomerase activity and thereby selectively inhibit cancer cell growth. The development of compounds with the necessary selectivity and affinity to target parallel-stranded G-quadruplex structures has proved particularly challenging to date, relying heavily upon limited structural data. We report here on a structure-based approach to the design of quadruplex-binding ligands to enhance affinity and selectivity for human telomeric DNA. Crystal structures have been determined of complexes between a 22-mer intramolecular human telomeric quadruplex and two potent tetra-substituted naphthalene diimide compounds, functionalized with positively charged N-methyl-piperazine side-chains. These compounds promote parallel-stranded quadruplex topology, binding exclusively to the 3' surface of each quadruplex. There are significant differences between the complexes in terms of ligand mobility and in the interactions with quadruplex grooves. One of the two ligands is markedly less mobile in the crystal complex and is more quadruplex-stabilizing, forming multiple electrostatic/hydrogen bond contacts with quadruplex phosphate groups. The data presented here provides a structural rationale for the biophysical (effects on quadruplex thermal stabilization) and biological data (inhibition of proliferation in cancer cell lines and evidence of in vivo antitumor activity) on compounds in this series and, thus, for the concept of telomere targeting with DNA quadruplex-binding small molecules.


Infantile colic, prolonged crying and maternal postnatal depression.
To study if infant crying is associated with maternal postnatal depression. Data from 1015 mothers and their children participating in a prospective European multicentre study were analysed. Infantile colic and prolonged crying were defined as excessive crying as reported by the mothers 2 and 6 months after delivery, and at the same time the mothers completed the Edinburgh Postnatal Depression Scale (EPDS). In cross-sectional analyses, infant crying was associated with high EPDS scores both 2 (OR: 4.4; 95% CI: 2.4-8.2) and 6 months postpartum (OR: 10.8; 95% CI: 4.3-26.9). More than one-third of the others of infants with prolonged crying had high EPDS scores 6 months postpartum. Longitudinal analyses showed that mothers of infants with colic had increased odds of having high EPDS scores 6 months after delivery even if crying had resolved (OR: 3.7; 95% CI: 1.4-10.1). Both infantile colic and prolonged crying were associated with high maternal depression scores. Most noteworthy, infantile colic at 2 months of age was associated with high maternal depression scores 4 months later.


Isoliquiritigenin Attenuates Atherogenesis in Apolipoprotein E-Deficient Mice.
Isoliquiritigenin (ISL) exhibits antioxidation and anti-inflammation activity. We sought to investigate the effects and mechanism of ISL on the development of atherosclerotic lesions in apolipoprotein E-deficient (apoE-/-) mice. Firstly, we determined that ISL reduced the mRNA levels of inflammatory factors interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), and monocyte chemotactic protein-1 (MCP-1), while it increased the expression of several lipoprotein-related genes in peritoneal macrophages treated with lipopolysaccharide (LPS). ISL also enhanced peroxisome proliferator-activated receptor gamma (PPARγ) protein levels and reversed the changes of ATP-binding cassette transporter A (ABCA1) and cluster of differentiation 36 (CD36) in macrophages treated with oxidative low-density lipoprotein (ox-LDL). Then, in an in vivo study, female apoE-/- mice were fed a Western diet with ISL (0, 20, 100 mg/kg/day) added for 12 weeks. We found that ISL decreased the plasma cholesterol levels of very low-density lipoprotein (VLDL)/LDL, promoted plasma superoxide dismutase (SOD) and paraoxonase-1 (PON1) activities, and decreased plasma IL-6, TNF-α, and MCP-1 levels. Moreover, ISL significantly reduced the atherosclerotic lesions and hepatic steatosis in apoE-/- mice. In the liver, ISL altered the expression of several key genes (such as SRBI, ABCA1, ABCG8, PPARγ, and FASN) involving cholesterol-selective uptake and excretion into bile, triglyceride (TG) biosynthesis, and inflammation. These results suggest that the atheroprotective effects of ISL are due to the improvement of lipid metabolism, antioxidation, and anti-inflammation, which involve PPARγ-dependent signaling.


The Scandinavian Stroke Scale is equally as good as The National Institutes of Health Stroke Scale in identifying 3-month outcome.
The National Institutes of Health Stroke Scale (NIHSS) is the first choice among stroke scales. The Scandinavian Stroke Scale (SSS) is an alternative scale that is easy to apply in the clinic. To compare the ability of the SSS with that of the NIHSS in identifying patients who are dead or dependent at 3-month follow-up. A prospective study including patients with acute stroke. NIHSS and SSS measurements were obtained during index hospital stay. The receiver operating characteristics curve was used to determine the optimal dichotomization of the NIHSS and the SSS by using a modified Rankin Scale (mRS) >2 at 3-month follow-up as the criterion standard. Positive and negative predictive values (PPV and NPV) were calculated. A total of 104 patients (mean age 79 years, 57.7% men) were included. Median (interquartile range (IQR)) NIHSS and SSS score were 6.0 (2.0-11.8) and 43.5 (30.0-51.0), respectively. The areas under the curve were 0.769 and 0.796 for NIHSS and SSS, respectively, χ2 (p = 0.303). The best cut-off point for NIHSS was 6/7 points (PPV = 76.2%, NPV = 69.0%) while for SSS it was 42/43 points (PPV = 71.4%, NPV = 73.2%). The SSS was as good as the NIHSS in identifying patients who had died or were dependent at 3-month follow-up. The measurement properties of the SSS should be investigated further.


Cyclooxygenase-2 catalysis and inhibition in lipid bilayer nanodiscs.
Cyclooxygenases (COX-1 and COX-2) oxygenate arachidonic acid (AA) to generate prostaglandins. The enzymes associate with one leaflet of the membrane bilayer. We utilized nanodisc technology to investigate the function of human (hu) COX-2 and murine (mu) COX-2 in a lipid bilayer environment. huCOX-2 and muCOX-2 were incorporated into nanodiscs composed of POPC, POPS, DOPC, or DOPS phospholipids. Size-exclusion chromatography and negative stain electron microscopy confirm that a single COX-2 homodimer is incorporated into the nanodisc scaffold. Nanodisc-reconstituted COX-2 exhibited similar kinetic profiles for the oxygenation of AA, eicosapentaenoic acid, and 1-arachidonoyl glycerol compared to those derived using detergent solubilized enzyme. Moreover, changing the phospholipid composition of the nanodisc did not alter the ability of COX-2 to oxygenate AA or to be inhibited by various nonselective NSAIDs or celecoxib. The cyclooxygenase activity of nanodisc-reconstituted COX-2 was reduced by aspirin acetylation and potentiated by the nonsubstrate fatty acid palmitic acid to the same extent as detergent solubilized enzyme, independent of phospholipid composition. The stabilization and maintenance of activity afforded by the incorporation of the enzyme into nanodiscs generates a native-like lipid bilayer environment to pursue studies of COX utilizing solution-based techniques that are otherwise not tractable in the presence of detergents.


Resistin as a potential marker of renal disease in lupus nephritis.
Systemic lupus erythematosus (SLE) and lupus nephritis (LN) have strong concomitance with cardiovascular disease that cannot be explained fully by typical risk factors. We examined the possibility that serum or urine expression of adipokines may act as biomarkers for LN, as these proteins have been associated previously with cardiovascular disease as well as SLE. Antibody arrays were performed on serum and urine from lupus patients and matched controls using a cross-sectional study design. From the initial array-based screening data of 15 adipokines, adiponectin, leptin and resistin were selected for validation by enzyme-linked immunosorbent assay (ELISA). Correlations were determined between adipokine expression levels and measures of disease activity or lupus nephritis. The expression of adiponectin and resistin was increased in both sera and urine from LN patients, while leptin was increased in LN patient sera, compared to matched controls. Serum resistin, but not urine resistin, was correlated with measures of renal dysfunction in LN. Serum resistin expression may be useful as a marker of renal dysfunction in patients with LN, although longitudinal studies are warranted. Further studies are necessary to determine if resistin has functional consequences in LN.


Hybrid lecture-online format increases student grades in an undergraduate exercise physiology course at a large urban university.
Hybrid courses allow students additional exposure to course content that is not possible in a traditional classroom environment. This exposure may lead to an improvement in academic performance. In this report, I describe the transition of a large undergraduate exercise physiology course from a traditional lecture format to a hybrid lecture-online format. A total of 658 final grades (traditional = 346, hybrid = 312) was used to evaluate the effect of course format on academic performance. The hybrid online portion was delivered using WebCT Vista, enhanced with various instructional technologies. The hybrid lecture portion was enhanced with an in-class response system. PowerPoint files were used to distribute in-class lectures in both formats of the course. Final student grades were 9.9% higher (83% of the increase due to an increase in the exam grade) when the course was administered in a hybrid format (P = 0.01), which translated to a one letter grade increase on a standard grading scale. Transition from a traditional lecture format to a hybrid format significantly enhanced student learning; presumably, this increase is due to the fact that students were able to increase their exposure to course content via access to material on WebCT.


Cesarean section in the setting of severe pulmonary hypertension requiring extracorporeal life support.
We describe the use of veno-arterial extracorporeal membrane oxygenation (ECMO) in a 35-year-old female with severe fixed pulmonary hypertension who went into cardiogenic shock during a Cesarean section. Pregnancy in the presence of severe pulmonary hypertension is typically contraindicated due to high maternal mortality rates. This patient visited our hospital at 37 weeks of gestation after experiencing dyspnea and chest pain. Clinical evaluation revealed severe fixed pulmonary hypertension. At the time of the planned delivery, femoral lines were placed; in case of emergency, ECMO became necessary during the delivery. During delivery, the patient developed sudden hemodynamic collapse necessitating rapid cannulation and initiation of ECMO. She was stabilized pharmacologically and separated from ECMO after 2 days. The baby was delivered uneventfully, and the mother and child were discharged 1 month after delivery.


Medicare in a consumer-choice environment: competitor or residual program?
The Medicare provisions of the reconciliation bill and President Clinton's proposals would both allow many new health plans to compete for Medicare enrollees and structure a competitive, consumer-choice system. Medicare will enter this new environment with serious shortcomings. The logic of market competition will call for upgrading many aspects of Medicare if it is to be successful. Among Medicare's features that need to be reconsidered are benefits, consumer focus, premium financing, payment policies, provider arrangements, performance accountability, management systems, risk adjustment and market strategies, social mission responsibilities, and research and development.


Transcription of a cloned Xenopus laevis H4 histone gene in the homologous frog oocyte system depends on an evolutionary conserved sequence motif in the -50 region.
A cloned Xenopus laevis H4 histone gene has been expressed in the X.laevis oocyte nucleus. The homologous histone H4 gene can be correctly and efficiently expressed in the frog oocyte even in presence of bacterial vector DNA. As revealed by both analytical gel electrophoresis and S1 mapping, two H4 mRNAs are specified with different transcriptional efficiencies from the tandemly repeated promoter. Results from deletion mapping of the sequences essential for promoting H4 transcription show that drastic reduction of transcription is obtained when the sequences lying between -64 and -35 bp from the mRNA cap site are removed. We demonstrate by DNA sequence comparison using a novel computer program that this important area of the H4 promoter contains two highly conserved DNA motifs near positions -51 to -46 upstream from the cap site in all H4 gene promoters analysed.


Cerebral regulatory T cells restrain microglia/macrophage-mediated inflammatory responses via IL-10.
Forkhead box P3 (Foxp3)(+) regulatory T (Treg) cells maintain the immune tolerance and prevent inflammatory responses in the periphery. However, the presence of Treg cells in the CNS under steady state has not been studied. Here, for the first time, we show a substantial TCRαβ (+) CD4(+) Foxp3(+) T-cell population (cerebral Treg cells) in the rat cerebrum, constituting more than 15% of the cerebral CD4(+) T-cell compartment. Cerebral Treg cells showed an activated/memory phenotype and expressed many Treg-cell signature genes at higher levels than peripheral Treg cells. Consistent with their activated/memory phenotype, cerebral Treg cells robustly restrained the LPS-induced inflammatory responses of brain microglia/macrophages, suggesting a role in maintaining the cerebral homeostasis by inhibiting the neuroinflammation. In addition, brain astrocytes were the helper cells that sustained Foxp3 expression in Treg cells through IL-2/STAT5 signaling, showing that the interaction between astrocytes and Treg cells contributes to the maintenance of Treg-cell identity in the brain. Taken together, our work represents the first study to characterize the phenotypic and functional features of Treg cells in the rat cerebrum. Our data have provided a novel insight for the contribution of Treg cells to the immunosurveillance and immunomodulation in the cerebrum under steady state.