Horizontal alveolar bone loss: A periodontal orphan.
Attempts to successfully regenerate lost alveolar bone have always been a clinician's dream. Angular defects, at least, have a fairer chance, but the same cannot be said about horizontal bone loss. The purpose of the present study was to evaluate the prevalence of horizontal alveolar bone loss and vertical bone defects in periodontal patients; and later, to correlate it with the treatment modalities available in the literature for horizontal and vertical bone defects. The study was conducted in two parts. Part I was the radiographic evaluation of 150 orthopantomographs (OPGs) (of patients diagnosed with chronic periodontitis and seeking periodontal care), which were digitized and read using the AutoCAD 2006 software. All the periodontitis-affected teeth were categorized as teeth with vertical defects (if the defect angle was ≤45° and defect depth was ≥3 mm) or as having horizontal bone loss. Part II of the study comprised search of the literature on treatment modalities for horizontal and vertical bone loss in four selected periodontal journals. Out of the 150 OPGs studied, 54 (36%) OPGs showed one or more vertical defects. Totally, 3,371 teeth were studied, out of which horizontal bone loss was found in 3,107 (92.2%) teeth, and vertical defects were found only in 264 (7.8%) of the teeth, which was statistically significant (P<.001). Search of the selected journals revealed 477 papers have addressed the treatment modalities for vertical and horizontal types of bone loss specifically. Out of the 477 papers, 461 (96.3%) have addressed vertical bone loss, and 18 (3.7%) have addressed treatment options for horizontal bone loss. Two papers have addressed both types of bone loss and are included in both categories. Horizontal bone loss is more prevalent than vertical bone loss but has been sidelined by researchers as very few papers have been published on the subject of regenerative treatment modalities for this type of bone loss. This study should be an impetus for greater attention to an otherwise ubiquitous periodontal challenge.


Interventions after acute stress prevent its delayed effects on the amygdala.
Stress is known to elicit contrasting patterns of plasticity in the amygdala and hippocampus. While chronic stress leads to neuronal atrophy in the rodent hippocampus, it has the opposite effect in the basolateral amygdala (BLA). Further, even a single episode of acute stress is known to elicit delayed effects in the amygdala. For example, 2 h of immobilisation stress has been shown to cause a delayed increase in dendritic spine density on BLA principal neurons 10 days later in young rats. This is paralleled by higher anxiety-like behaviour at the same delayed time point. This temporal build-up of morphological and behavioural effects 10 days later, in turn, provides a stress-free time window of intervention after exposure to acute stress. Here, we explore this possibility by specifically testing the efficacy of an anxiolytic drug in reversing the delayed effects of acute immobilisation stress. Oral gavage of diazepam 1 h after immobilisation stress prevented the increase in anxiety-like behaviour on the elevated plus-maze 10 days later. The same post-stress intervention also prevented delayed spinogenesis in the BLA 10 days after acute stress. Surprisingly, gavage of only the vehicle also had a protective effect on both the behavioural and synaptic effects of stress 10 days later. Vehicle gavage was found to trigger a significant rise in corticosterone levels that was comparable to that elicited by acute stress. This suggests that a surge in corticosterone levels, caused by the vehicle gavage 1 h after acute stress, was capable of reversing the delayed enhancing effects of stress on anxiety-like behaviour and BLA synaptic connectivity. These findings are consistent with clinical reports on the protective effects of glucocorticoids against the development of symptoms of post-traumatic stress disorder. Taken together, these results reveal strategies, targeted 1 h after stress, which can prevent the delayed effects of a brief exposure to a severe physical stressor.


A prospective study to validate the functional assessment of cancer therapy (FACT) for epidermal growth factor receptor inhibitor (EGFRI)-induced dermatologic toxicities FACT-EGFRI 18 questionnaire: SWOG S1013.
Papulopustular rash is a common class effect of epidermal growth factor receptor inhibitors (EGFRI) that can affect patients' health-related quality of life and cause disruptions to treatment. SWOG S1013 (NCT01416688) is a multi-center study designed to validate the Functional Assessment of Cancer Therapy EGFRI 18 (FACT-EGFRI 18) using 7-items from the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) version 4.0 to assess EGFRI-induced skin-related toxicities and their impact on functional status. Patients with a diagnosis of colorectal or lung cancer to receive EGFRI therapies for at least 6 weeks were enrolled. Patient self-assessments using the FACT-EGFRI 18 were completed prior to undergoing CTCAE assessment by trained clinicians at baseline, weekly × 6, and then monthly × 3. The psychometric properties of the FACT-EGFRI 14 (skin toxicity items only) and 18 (plus 2 nail and 2 hair items) were established based on criterion validity, known groups validity, internal consistency reliability, and responsiveness to change. Of the 146 registered patients, 124 were evaluable. High Cronbach's alpha (> 0.70) for both FACT-EGFRI 14 and FACT-EGFRI 18 scores across assessment times were observed. Although agreement (i.e. criterion validity) between individual and summary scales of the FACT-EGFRI 18 for assessing skin toxicity was good, agreement with the clinician-reported CTCAE was only fair. The minimal important difference was determined to be 3 points. The results also demonstrated responsiveness to symptom change. Based on the results of this multi-center validation study, the FACT-EGFRI 18 patient-reported outcome instrument provided data from the patient's perspective yielding unique information as well as complementing clinician-rated CTCAE grades, especially for the symptoms of pain, pruritus, and paronychia. Good to excellent psychometric properties for the FACT-EGFRI 18 were demonstrated, supporting further use of this patient-reported outcomes measure. Additional validation with a more diverse group of patients should be conducted.


Feeling loved and integrated or lonely and rejected in everyday life: The role of age and social motivation.
Social approach and social avoidance goals (i.e., approach of positive and avoidance of negative outcomes in social situations) are important predictors of the feeling of being socially integrated or isolated. However, little is known about the development of these goals across adulthood. In a large diary study with N = 744 young (18-39 years), middle-aged (40-59 years), and older adults (60-83 years), we tested the hypothesis that the adaptiveness of social goals changes across adulthood: Social approach goals were hypothesized to be adaptive during young adulthood when adult social relationships are to be established. In contrast, social avoidance goals were hypothesized to become more adaptive with age as people are increasingly motivated to avoid interpersonal tension. Our findings support these hypotheses: Social approach goals were positively and social avoidance goals negatively associated with younger but not with middle-aged and older adults' daily social well-being. These results were robust across different situations (positive, negative) and different types of relationships (close, peripheral). The study highlights the changing role of social approach and avoidance goals for daily social well-being across adulthood. (PsycINFO Database Record


Swelling of interpenetrating networklike particles in a soft polymer matrix.
Fast transient fluorescence technique was used to monitor swelling of hard latex particles in a soft polymer matrix. Various film samples were prepared from pyrene (P) and/or naphthalene (N)-labeled poly(methyl methacrylate) (PMMA) particles in a low-molecular-weight poly(isobutylene) (PIB) matrix. These PMMA particles contain interpenetrating PIB channels. Film samples were annealed at elevated temperatures to promote particle swelling. Fluorescence lifetimes, tau, were measured for each film sample. It was observed that tau values decrease as the PIB content in the matrix is increased. The decrease in tau was explained by the increase in quenching of excited P and N molecules by low-molecular-weight PIB penetrating into the PIB channels in the PMMA particles. A drastic decrease in tau above a certain temperature was attributed to the effect of particle size on the swelling of the latex particles in the PIB matrix. It was observed that small particles (P-labeled) swell at much lower annealing temperatures than large (N-labeled) particles.


Long-term oxytocin administration improves social behaviors in a girl with autistic disorder.
Patients with autism spectrum disorders (ASDs) exhibit core autistic symptoms including social impairments from early childhood and mostly show secondary disabilities such as irritability and aggressive behavior based on core symptoms. However, there are still no radical treatments of social impairments in these patients. Oxytocin has been reported to play important roles in multiple social behaviors dependent on social recognition, and has been expected as one of the effective treatments of social impairments of patients with ASDs. We present a case of a 16-year-old girl with autistic disorder who treated by long-term administration of oxytocin nasal spray. Her autistic symptoms were successfully treated by two month administration; the girl's social interactions and social communication began to improve without adverse effects. Her irritability and aggressive behavior also improved dramatically with marked decreases in aberrant behavior checklist scores from 69 to 7. This case is the first to illustrate long-term administration of oxytocin nasal spray in the targeted treatment of social impairments in a female with autistic disorder. This case suggests that long-term nasal oxytocin spray is promising and well-tolerated for treatment of social impairments of patients with ASDs.


An overview of psychological and social factors in Charles Bonnet syndrome.
Charles Bonnet syndrome (CBS) is a condition where cognitively normal individuals with sight impairment experience simple and/or complex visual hallucinations. The exact pathogenesis of CBS is unknown; however, deafferentation is often recognised as a causal mechanism. Studies have provided insight into the multifaceted impact of CBS on wellbeing. Onset of CBS may cause distress among those believing visual hallucinations are indicative of a neurological condition. Hallucinatory content is often congruent with the emotional response. For example, hallucinations of a macabre nature typically result in a fearful response. Visual hallucinations may be highly disruptive, causing everyday tasks to become challenging. Clinical management relies on forewarning and pre-emptive questioning. Yet, knowledge and awareness of CBS is typically low. In this review, we provide a summary of the social and psychological implications of CBS and explore recent developments aimed at raising awareness and improving patient management.


Asparagine endopeptidase is required for normal kidney physiology and homeostasis.
Although protein recapture and catabolism is known as a key function of kidney proximal tubular cells (PTCs), to date, no single protease has been shown to be required. Asparagine endopeptidase (AEP) is an unusually specific endosomal and lysosomal cysteine protease, expressed at high levels in the PTCs of the mammalian kidney. We report that mice lacking AEP accumulate a discrete set of proteins in their PTC endosomes and lysosomes, which indicates a defect in the normal catabolism of proteins captured from the filtrate. Moreover, the mice develop progressive kidney pathology, including hyperplasia of PTCs, interstitial fibrosis, development of glomerular cysts, and renal pelvis dilation. By 6 mo of age, the glomerular filtration rate in AEP-null mice dropped by almost a factor of 2, and the mice developed proteinuria. We also show that EGF receptor levels are significantly higher in AEP-null PTCs, which likely explains the hyperplasia, and we show that chemical inhibition of AEP activity suppresses down-regulation of the EGF receptor in vitro. Thus, AEP is required for normal protein catabolism by PTCs, and its loss induces proliferative and other abnormalities in the murine kidney, at least in part through defective regulation of the EGF receptor.


Monocyte Subsets: Phenotypes and Function in Tuberculosis Infection.
Monocytes are critical defense components that play an important role in the primary innate immune response. The heterogeneous nature of monocytes and their ability to differentiate into either monocyte-derived macrophages or monocyte-derived dendritic cells allows them to serve as a bridge between the innate and adaptive immune responses. Current studies of monocytes based on immunofluorescence, single-cell RNA sequencing and whole mass spectrometry finger printing reveals different classification systems for monocyte subsets. In humans, three circulating monocyte subsets are classified based on relative expression levels of CD14 and CD16 surface proteins, namely classical, intermediate and non-classical subsets. Transcriptomic analyses of these subsets help to define their distinct functional properties. Tuberculosis (TB) is a disease instigated by the deadly pathogen Mycobacterium tuberculosis. Current research on monocytes in TB has indicated that there are alterations in the frequency of intermediate and non-classical subsets suggesting their impact in bacterial persistence. In this review, we will focus on these monocyte subsets, including their classification, frequency distribution, cytokine profiles, role as a biomarker and will comment on future directions for understanding the salient phenotypic and functional properties relevant to TB pathogenesis.


Persistent hyperreactivity and reactive airway dysfunction in firefighters at the World Trade Center.
New York City Fire Department rescue workers experienced massive exposure to airborne particulates at the World Trade Center site. Aims of this longitudinal study were to (1) determine if bronchial hyperreactivity was present, persistent, and independently associated with exposure intensity, (2) identify objective measures shortly after the collapse that would predict persistent hyperreactivity and a diagnosis of reactive airways dysfunction 6 months post-collapse. A representative sample of 179 rescue workers stratified by exposure intensity (high, moderate, and control) without current smoking or prior respiratory disease was enrolled. Highly exposed workers arrived within 2 hours of collapse, moderately exposed workers arrived later on Days 1-2; control subjects were not exposed. Hyperreactivity at 1, 3, and 6 months post-collapse was associated with exposure intensity, independent of ex-smoking and airflow obstruction. Six months post-collapse, highly exposed workers were 6.8 times more likely than moderately exposed workers and control subjects to be hyperreactive (95% confidence interval, 1.8-25.2; p = 0.004), and hyperreactivity persisted in 55% of those hyperreactive at 1 and/or 3 months. In highly exposed subjects, hyperreactivity 1 or 3 months post-collapse was the sole predictor for reactive airways dysfunction (p = 0.021). In conclusion, development and persistence of hyperreactivity and reactive airways dysfunction were strongly and independently associated with exposure intensity. Hyperreactivity shortly post-collapse predicted reactive airways dysfunction at 6 months in highly exposed workers; this has important implications for disaster management.


Email for clinical communication between patients/caregivers and healthcare professionals.
Email is a popular and commonly-used method of communication, but its use in health care is not routine. Where email communication has been demonstrated in health care this has included its use for communication between patients/caregivers and healthcare professionals for clinical purposes, but the effects of using email in this way is not known.This review addresses the use of email for two-way clinical communication between patients/caregivers and healthcare professionals. To assess the effects of healthcare professionals and patients using email to communicate with each other, on patient outcomes, health service performance, service efficiency and acceptability. We searched: the Cochrane Consumers and Communication Review Group Specialised Register, Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 1 2010), MEDLINE (OvidSP) (1950 to January 2010), EMBASE (OvidSP) (1980 to January 2010), PsycINFO (OvidSP) (1967 to January 2010), CINAHL (EbscoHOST) (1982 to February 2010) and ERIC (CSA) (1965 to January 2010). We searched grey literature: theses/dissertation repositories, trials registers and Google Scholar (searched July 2010). We used additional search methods: examining reference lists, contacting authors. Randomised controlled trials, quasi-randomised trials, controlled before and after studies and interrupted time series studies examining interventions using email to allow patients to communicate clinical concerns to a healthcare professional and receive a reply, and taking the form of 1) unsecured email 2) secure email or 3) web messaging. All healthcare professionals, patients and caregivers in all settings were considered. Two authors independently assessed the risk of bias of included studies and extracted data. We contacted study authors for additional information. We assessed risk of bias according to the Cochrane Handbook for Systematic Reviews of Interventions. For continuous measures, we report effect sizes as mean differences (MD). For dichotomous outcome measures, we report effect sizes as odds ratios and rate ratios. Where it was not possible to calculate an effect estimate we report mean values for both intervention and control groups and the total number of participants in each group. Where data are available only as median values it is presented as such. It was not possible to carry out any meta-analysis of the data. We included nine trials enrolling 1733 patients; all trials were judged to be at risk of bias. Seven were randomised controlled trials; two were cluster-randomised controlled designs. Eight examined email as compared to standard methods of communication. One compared email with telephone for the delivery of counselling. When email was compared to standard methods, for the majority of patient/caregiver outcomes it was not possible to adequately assess whether email had any effect. For health service use outcomes it was not possible to adequately assess whether email has any effect on resource use, but some results indicated that an email intervention leads to an increased number of emails and telephone calls being received by healthcare professionals. Three studies reported some type of adverse event but it was not clear if the adverse event had any impact on the health of the patient or the quality of health care. When email counselling was compared to telephone counselling only patient outcomes were measured, and for the majority of measures there was no difference between groups. Where there were differences these showed that telephone counselling leads to greater change in lifestyle modification factors than email counselling. There was one outcome relating to harm, which showed no difference between the email and the telephone counselling groups. There were no primary outcomes relating to healthcare professionals for either comparison. The evidence base was found to be limited with variable results and missing data, and therefore it was not possible to adequately assess the effect of email for clinical communication between patients/caregivers and healthcare professionals. Recommendations for clinical practice could not be made. Future research should ideally address the issue of missing data and methodological concerns by adhering to published reporting standards. The rapidly changing nature of technology should be taken into account when designing and conducting future studies and barriers to trial development and implementation should also be tackled. Potential outcomes of interest for future research include cost-effectiveness and health service resource use.


Comparison between Flow Cytometry, Microscopy, and Lactate Dehydrogenase-Based Enzyme-Linked Immunosorbent Assay for Plasmodium falciparum Drug Susceptibility Testing under Field Conditions.
Flow cytometry is an objective method for conducting in vitro antimalarial sensitivity assays with increasing potential for application in field sites. We examined in vitro susceptibility to seven anti-malarial drugs for 40 fresh P. falciparum field isolates via a flow cytometry method (FCM), a colorimetric LDH-based ELISA : DELI), and standard microscopic slide analysis of growth. For FCM, 184/280 (66%) assays met analytical acceptance criteria, compared to 166/280 (59%) for DELI. There was good agreement between FCM and microscopy, while DELI tended to produce higher half-maximal inhibition constants (IC50s) than FCM, with an overall bias of 2.2-fold (Bland-Altman comparison). Values for artesunate and dihydroartemisinin were most affected. Paradoxical increases in signal at very high concentrations of mefloquine and related compounds were more marked with the DELI assay, suggesting that off-target effects on LDH production may be responsible. Loss of FCM signal due to reinvasion or slow growth was observed in a small number of samples. These results extend previous work on use of flow cytometry to determine antimalarial susceptibility in terms of the number of samples, range of drugs, and comparison with other methods.


Assessing a new approach to verbal autopsy interpretation in a rural Ethiopian community: the InterVA model.
Verbal autopsy (VA) -- the interviewing of family members or caregivers about the circumstances of a death after the event -- is an established tool in areas where routine death registration is non-existent or inadequate. We assessed the performance of a probabilistic model (InterVA) for interpreting community-based VA interviews, in order to investigate patterns of cause-specific mortality in a rural Ethiopian community. We compared results with those obtained after review of the VA by local physicians, with a view to validating the model as a community-based tool. Two-hundred and eighty-nine VA interviews were successfully completed; these included most deaths occurring in a defined community over a 1-year period. The VA interviews were interpreted by physicians and by the model, and cause-specific mortality fractions were derived for the whole community and for particular age groups using both approaches. The results of the two approaches to interpretation correlated well in this example from Ethiopia. Four major cause groups accounted for over 60% of all mortality, and patterns within specific age groups were consistent with expectations for an underdeveloped high-mortality community in sub-Saharan Africa. Compared with interpretation by physicians, the InterVA model is much less labour intensive and offers 100% consistency. It is a valuable new tool for characterizing patterns of cause-specific mortality in communities without death registration and for comparing patterns of mortality in different populations.


The effects of hyperglycaemic hypoxia on rectification in rat dorsal root axons.
1. Electrotonic responses to 150 ms current pulses were recorded from isolated rat dorsal roots incubated for at least 3 h with either normal (5 mM) or high (25 mM) D-glucose solutions, and with either normal (25 mM) or low (5 mM) bicarbonate concentrations. 2. On replacement of O2 by N2 for 50 min, all the roots depolarized, but the changes in electrotonus differed systematically. With normal glucose, the depolarization was accompanied by an increase in input conductance. In contrast, for the hyperglycaemic roots the depolarization was slower and accompanied by a fall in input conductance which was exacerbated in low bicarbonate concentrations. 3. The changes induced by hyperglycaemic hypoxia in low bicarbonate could be mimicked by exposure of the roots either to 100% CO2 or to a combination of 3 mM tetraethylammonium chloride and 3 mM 4-aminopyridine, to block both fast and slow potassium channels. 4. These results indicate that the primary mechanism of hypoxic depolarization of these sensory axons is altered by hyperglycaemia. In normoglycaemia, the changes in electrotonus are consistent with an increase in axonal potassium conductance. The block of potassium channels seen in hyperglycaemic hypoxia is attributed to intra-axonal acidification by anaerobic glycolysis and may contribute to the pathogenesis of diabetic neuropathy.


Evaluating the Psychometric Properties of the 7-Item Persian Game Addiction Scale for Iranian Adolescents.
The 7-item Gaming Addiction Scale (GAS) is a brief instrument based on DSM criteria to assess gaming addiction. Although the psychometric properties of the GAS have been tested using classical test theory, its psychometric properties have never been tested using modern test theory (e.g., Rasch analysis). The present study used a large adolescent sample in Iran to test the psychometric properties of the Persian GAS through both classical test and modern test theories. Adolescents (n = 4442; mean age = 15.3 years; 50.3% males) were recruited from Qazvin, Iran. In addition to the GAS, all of them completed the following instruments: the nine-item Internet Gaming Disorder Scale-Short Form (IGDS-SF9), Depression Anxiety Stress Scale (DASS), Pittsburgh Sleep Quality Index (PSQI), and a generic quality of life instrument. Two weeks later, all participants completed the GAS again. Confirmatory factor analysis (CFA) and Rasch analysis were used to test the unidimensionality of the GAS. Pearson correlation coefficients were used to test the test-retest reliability, and a regression model was used to test the criterion-related validity of the GAS. Both CFA and Rasch analysis supported the unidimensionality of the GAS. Pearson correlations coefficients showed satisfactory test-retest reliability of the GAS (r = 0.78 to 0.86), and the regression model demonstrated the criterion-related validity of the GAS (β = 0.31 with IGDS-SF9; 0.41 with PSQI). Based on the results, the Persian GAS is a reliable and valid instrument for healthcare providers to assess the level of gaming addiction among Persian-speaking adolescents.


Role of glycerol 3-phosphate dehydrogenase in glyceride metabolism. Effect of diet on enzyme activities in chicken liver.
1. The metabolic role of hepatic NAD-linked glycerol 3-phosphate dehydrogenase (EC 18.104.22.168) was investigated vis-a-vis glyceride synthesis, glyceride degradation and the maintainence of the NAD redox state. 2. Five-week-old chickens were placed on five dietary regimes: a control group, a group on an increased-carbohydrate-lowered-fat diet, a group on a high-fat-lowered-carbohydrate diet, a starved group and a starved-refed group. In each group the specific activity (mumol/min per g wet wt. of tissue) of hepatic glycerol 3-phosphate dehydrogenase was compared with the activities of the beta-oxoacyl-(acyl-carrier protein) reductase component of fatty acid synthetase, glycerol kinase (EC 188.8.131.52) and lactate dehydrogenase (EC 18.104.22.168). 3. During starvation, the activities of glycerol 3-phosphate dehydrogenase, glycerol kinase and lactate dehydrogenase rose significantly. After re-feeding these activities returned to near normal. All three activities rose slightly on the high-fat diet. Lactate dehydrogenase activity rose slightly, whereas those of the other two enzymes fell slightly on the increased-carbohydrate-lowered-fat diet. 4. The activity of the beta-oxoacyl-(acyl-carrier protein) reductase component of fatty acid synthetase, a lipid-synthesizing enzyme, contrasted strikingly with the other three enzyme activities. Its activity was slightly elevated on the increased-carbohydrate diet and significantly diminished on the high-fat diet and during starvation. 5. The changes in activity of the chicken liver isoenzyme of glycerol 3-phosphate dehydrogenase in response to dietary stresses suggest that the enzyme has an important metabolic role other than or in addition to glyceride biosynthesis.


The LifeLines Cohort Study: Prevalence and treatment of cardiovascular disease and risk factors.
The LifeLines Cohort Study is a large three-generation prospective study and Biobank. Recruitment and data collection started in 2006 and follow-up is planned for 30years. The central aim of LifeLines is to understand healthy ageing in the 21st century. Here, the study design, methods, baseline and major cardiovascular phenotypes of the LifeLines Cohort Study are presented. Baseline cardiovascular phenotypes were defined in 9700 juvenile (8-18years) and 152,180 adult (≥18years) participants. Cardiovascular disease (CVD) was defined using ICD-10 criteria. At least one cardiovascular risk factor was present in 73% of the adult participants. The prevalence, adjusted for the Dutch population, was determined for risk factors (hypertension (33%), hypercholesterolemia (19%), diabetes (4%), overweight (56%), and current smoking (19%)) and CVD (myocardial infarction (1.8%), heart failure (1.0%), and atrial fibrillation (1.3%)). Overall CVD prevalence increased with age from 9% in participants<65years to 28% in participants≥65years. Of the participants with hypertension, hypercholesterolemia and diabetes, respectively 75%, 96% and 41% did not receive preventive pharmacotherapy. The contemporary LifeLines Cohort Study provides researchers with unique and novel opportunities to study environmental, phenotypic, and genetic risk factors for CVD and is expected to improve our knowledge on healthy ageing. In this contemporary Western cohort we identified a remarkable high percentage of untreated CVD risk factors suggesting that not all opportunities to reduce the CVD burden are utilised.


MUMPS MYOCARDITIS: A FORGOTTEN DISEASE?
Mumps is an acute viral illness that follows a self-limiting course but up to 10% of cases have a complicated course with the involvement of other organ systems. Myocarditis is reported as a complication but the incidence has greatly fallen ever since the development of the mumps vaccine. A child presented to our department with parotid swelling and fever. Persistent tachycardia with irregular pulse led to further cardiac work up which showed decreased ejection fraction and raised serum cardiac enzymes, indicating myocardial damage. With ionotropic agents and supportive care, there was complete normalization of ejection fraction and serum cardiac enzyme levels. He was discharged within a week of admission. This case highlights the importance of suspecting myocarditis in the setting of mumps, a diagnosis that precludes early suspicion in mumps patients suffering from cardiac symptoms not explained by other potential aetiologies. Early suspicion and timely supportive care are essential to ensure favourable outcomes.


[A fresh look at psychiatric disorders].
The a-theoretical approach to psychiatric disorders, introduced via dsm iii, has had a tremendous impact. It has stimulated a large body of research, facilitated by the concurrent development of new techniques in genetics, neuro-imaging and neuropsychology. However, the research results of the last twenty years or so have cast doubt on the validity of the clinical categories set out in dsm iii. To develop a new view on developmental pathways in psychopathology, clinical assessment and scientifically acceptable classification. In this article we review the state of the art with regard to underlying endophenotypes at the level of brain and neurotransmitter functioning and neuropsychology and we consider the effect of social determinants on the developments of psychopathology. Our results show that neither genotypes and endophenotypes, nor brain mechanism, nor neuropsychological deviances have a one-to-one correlation with clinical categories as defined in even the dsm 5. dsm-5 provides a range of possibilities for classifying psychiatric disorders at symptom level. But these categories seem to be less distinct than was at first assumed. Recent research has shown that there is a great deal of overlap at the genetic, epigenetic and endophenotype level. This calls for more emphasis on individual assessment and diagnostics in both clinical practice and scientific research. More attention needs to be given to the dimensions of emotion and behavior, vulnerability and resilience. This type of approach, involving genotypes, endophenotypes, epigenetics and brain functioning, could help to elucidate the interaction between these various levels and/or explain the underlying mechanisms of psychiatric disorders.


Extracellular vesicles from recombinant cell factories improve the activity and efficacy of enzymes defective in lysosomal storage disorders.
In the present study the use of extracellular vesicles (EVs) as vehicles for therapeutic enzymes in lysosomal storage disorders was explored. EVs were isolated from mammalian cells overexpressing alpha-galactosidase A (GLA) or N-sulfoglucosamine sulfohydrolase (SGSH) enzymes, defective in Fabry and Sanfilippo A diseases, respectively. Direct purification of EVs from cell supernatants was found to be a simple and efficient method to obtain highly active GLA and SGSH proteins, even after EV lyophilization. Likewise, EVs carrying GLA (EV-GLA) were rapidly uptaken and reached the lysosomes in cellular models of Fabry disease, restoring lysosomal functionality much more efficiently than the recombinant enzyme in clinical use. In vivo, EVs were well tolerated and distributed among all main organs, including the brain. DiR-labelled EVs were localized in brain parenchyma 1 h after intra-arterial (internal carotid artery) or intravenous (tail vein) administrations. Moreover, a single intravenous administration of EV-GLA was able to reduce globotriaosylceramide (Gb3) substrate levels in clinically relevant tissues, such kidneys and brain. Overall, our results demonstrate that EVs from cells overexpressing lysosomal enzymes act as natural protein delivery systems, improving the activity and the efficacy of the recombinant proteins and facilitating their access to organs neglected by conventional enzyme replacement therapies.


Listening to speech in a background of other talkers: effects of talker number and noise vocoding.
Some of the most common interfering background sounds a listener experiences are the sounds of other talkers. In Experiment 1, recognition for natural Institute of Electrical and Electronics Engineers (IEEE) sentences was measured in normal-hearing adults at two fixed signal-to-noise ratios (SNRs) in 16 backgrounds with the same long-term spectrum: unprocessed speech babble (1, 2, 4, 8, and 16 talkers), noise-vocoded versions of the babbles (12 channels), noise modulated with the wide-band envelope of the speech babbles, and unmodulated noise. All talkers were adult males. For a given number of talkers, natural speech was always the most effective masker. The greatest changes in performance occurred as the number of talkers in the maskers increased from 1 to 2 or 4, with small changes thereafter. In Experiment 2, the same targets and maskers (1, 2, and 16 talkers) were used to measure speech reception thresholds (SRTs) adaptively. Periodicity in the target was also manipulated by noise-vocoding, which led to considerably higher SRTs. The greatest masking effect always occurred for the masker type most similar to the target, while the effects of the number of talkers were generally small. Implications are drawn with reference to glimpsing, informational vs energetic masking, overall SNR, and aspects of periodicity.


Physico-chemical controls on phosphorus cycling in two lowland streams. Part 2--the sediment phase.
This article investigates the temporal and spatial controls on sediment-phosphorus (P) dynamics in two contrasting sub-catchments of the River Kennet, England. Suspended sediment (collected under representative flow conditions) and size-fractionated bedload (collected weekly for one year) from the Rivers Lambourn and Enborne was analysed for a range of physico-chemical determinands. Total P concentrations were highest in the most mobile fractions of sediment: suspended sediment, fine silt and clay and organic matter (mean concentrations of 1758, 1548 and 1440 microg P g(-1) dry sediment, respectively). Correlation analysis showed significant relationships between total P and total iron (n= 110), total manganese (n= 110), organic matter (n= 110) and specific surface area (n= 28) in the Lambourn (r2 0.71, 0.68, 0.62 and 0.52, respectively) and between total P and total iron (n= 110), total manganese (n= 110) and organic matter (n= 110) in the Enborne (r2 0.74, 0.85 and 0.68, respectively). These data highlight the importance of metal oxyhydroxide adsorption of P on fine particulates and organic matter. However, high total P concentrations in the granule gravel and coarse sand size fraction during the summer period (mean concentration 228 microg P g(-1) dry sediment) also highlight the role of calcite co-precipitation on P dynamics in the Lambourn. P to cation ratios in Lambourn sediment indicated that fine silt and clay and granule gravel and coarse sand size fractions were potential sources of P release to the water column during specific periods of the summer and autumn. In the Enborne, however, only the granule gravel and coarse sand size fraction had high ratios and a slow, constant release of P was observed. In addition, scanning electron microscopy work confirmed the association of P with calcite in the Lambourn and P with iron on clay particles in the Enborne. The study highlighted the importance of the chemical and physical properties of the sediment in influencing the mechanisms controlling P storage and release within river channels.


Evaluation for safety of cultured corneal fibroblasts with cotreatment of alcohol and mitomycin C.
To investigate the effects of alcohol and mitomycin C (MMC) on cultured corneal fibroblast of the rabbit to determine the safety of this compound for clinical use. Corneal fibroblasts of New Zealand rabbits were cultured. Various concentrations (0%, 10%, 20%, 30%, 40%, and 60%) of ethanol were applied for 10, 20, 30, and 40 seconds to estimate dose- and time-dependent responses of cultured corneal fibroblasts. Cell viability was assessed using the MTT assay. Treated cells were additionally stained with Hoechst and annexin V for the identification of apoptosis. To investigate the coeffects of ethanol and MMC, dose and time dependency were evaluated after treatment with various concentrations of ethanol and MMC at different exposure times, and cell viability was established. To determine the latent effects of ethanol and MMC, cultured corneal fibroblasts were cotreated with various concentrations of ethanol and 0.02% MMC for various periods and washed out, and then one group was incubated for 24 hours and another group was not incubated. Cell viability was estimated, and Hoechst and annexin V staining were performed before and after incubation. To establish the pathway of cell death, caspase-3 activities were measured in cultured corneal fibroblasts treated with ethanol or MMC. Cell viability after ethanol treatment was dose and time dependent. After application of ethanol for 10 seconds, cell viability was significantly reduced with 20% ethanol (P = 0.001). At 20, 30, and 40 seconds of treatment with 10% ethanol, cell viability was significantly reduced (P < 0.01). Hoechst and annexin V staining revealed typical characteristics of apoptosis, such as bright fluorescent chromatin condensation, low fluorescence of nuclear fragmentation, and cell membrane shrinkage. Cell viability was more significantly reduced after cotreatment with alcohol and MMC, compared with treatment with alcohol alone. Moreover, cell viability was considerably decreased in the incubated group, compared with the nonincubated group. After 24 hours of incubation, cultured corneal fibroblasts cotreated with 10% ethanol and 0.02% MMC were stained with Hoechst and annexin V. Results were similar to data obtained with ethanol-treated cells. However, after application of 20% alcohol and MMC, a significant number of cells were not viable and were detached from the well walls. Caspase-3 activity significantly increased after treatment with 30% ethanol only and 30% ethanol in conjunction with 0.02% MMC. Alcohol and MMC reduced cell viability in cultured corneal fibroblasts in a dose- and time-dependent manner and had synergistic effects. This is related to the caspase-3 pathway, especially with concentrations of ethanol over 30%. Cotreatment with these reagents may significantly damage cultured corneal fibroblasts.


A modular analysis of the auxin signalling network.
Auxin is essential for plant development from embryogenesis onwards. Auxin acts in large part through regulation of transcription. The proteins acting in the signalling pathway regulating transcription downstream of auxin have been identified as well as the interactions between these proteins, thus identifying the topology of this network implicating 54 Auxin Response Factor (ARF) and Aux/IAA (IAA) transcriptional regulators. Here, we study the auxin signalling pathway by means of mathematical modeling at the single cell level. We proceed analytically, by considering the role played by five functional modules into which the auxin pathway can be decomposed: the sequestration of ARF by IAA, the transcriptional repression by IAA, the dimer formation amongst ARFs and IAAs, the feedback loop on IAA and the auxin induced degradation of IAA proteins. Focusing on these modules allows assessing their function within the dynamics of auxin signalling. One key outcome of this analysis is that there are both specific and overlapping functions between all the major modules of the signaling pathway. This suggests a combinatorial function of the modules in optimizing the speed and amplitude of auxin-induced transcription. Our work allows identifying potential functions for homo- and hetero-dimerization of transcriptional regulators, with ARF:IAA, IAA:IAA and ARF:ARF dimerization respectively controlling the amplitude, speed and sensitivity of the response and a synergistic effect of the interaction of IAA with transcriptional repressors on these characteristics of the signaling pathway. Finally, we also suggest experiments which might allow disentangling the structure of the auxin signaling pathway and analysing further its function in plants.


Method for the reduction of signal-induced noise in photomultiplier tubes.
A new method to reduce photomultiplier tube detector signal-induced noise (SIN) in a lidar system is successfully demonstrated. A metal ring electrode placed external to the photomultiplier tube photocathode is pulsed during the intense near-field lidar return with a potential between 15 and 500 V, resulting in a significant reduction in SIN. The effect of the metal ring voltage on the decay time constant and the magnitude of a simulated lidar signal is presented. Optimal experimental conditions for the use of this device in lidar receivers, such that the lidar decay time constant is not affected, are determined. Mechanisms for this SIN suppression system are discussed in detail, and data were recorded to show that the voltage on the metal ring functions by altering the photomultiplier electron optics.


Use of dual protection among female sex workers in Swaziland.
Female sex workers are at heightened risk of both HIV infection and unwanted pregnancy. Nonbarrier modern contraceptives are highly effective at preventing pregnancy, but offer no HIV protection. A better understanding of sex workers' use of condoms and nonbarrier methods is needed to help them meet their contraceptive and STI protection needs. A 2011 respondent-driven sampling survey collected reproductive health and contraceptive use data from 325 female sex workers in Swaziland. Multinomial logistic regression analysis was used to identify associations between selected characteristics and four outcomes of contraceptive use over the past month: consistent condom use alone; nonbarrier modern contraceptive use (either alone or with inconsistent condom use); dual method use; and inconsistent condom use, other method use or nonuse. Adjusted predicted probabilities were also calculated to determine patterns of association. After adjustments were made for background and behavioral factors, 16% of female sex workers were found to be consistent users of condoms alone; 39% used nonbarrier modern methods (without consistent condom use); 8% were dual method users; and 38% were inconsistent condom users or used other methods or none. Women who reported recent condom failure were less likely than others to be consistent condom users (6% vs. 22%). Consistent use of condoms alone was more common among women who had had no noncommercial partners in the past month than among those who reported two or more such partners (39% vs. 3%). In addition, respondents who had children were more likely than their nulliparous counterparts to report use of nonbarrier methods alone (65% vs. 14%). Inconsistent or no condom use among nonbarrier contraceptive users underscores the need to incorporate HIV prevention into family planning interventions, particularly among female sex workers who have children and noncommercial partners.


Using a logic model to evaluate the Kids Together early education inclusion program for children with disabilities and additional needs.
Despite clear evidence that learning and social opportunities for children with disabilities and special needs are more effective in inclusive not segregated settings, there are few known effective inclusion programs available to children with disabilities, their families or teachers in the early years within Australia. The Kids Together program was developed to support children with disabilities/additional needs aged 0-8 years attending mainstream early learning environments. Using a key worker transdisciplinary team model, the program aligns with the individualised package approach of the National Disability Insurance Scheme (NDIS). This paper reports on the use of a logic model to underpin the process, outcomes and impact evaluation of the Kids Together program. The research team worked across 15 Early Childhood Education and Care (ECEC) centres and in home and community settings. A realist evaluation using mixed methods was undertaken to understand what works, for whom and in what contexts. The development of a logic model provided a structured way to explore how the program was implemented and achieved short, medium and long term outcomes within a complex community setting. Kids Together was shown to be a highly effective and innovative model for supporting the inclusion of children with disabilities/additional needs in a range of environments central for early childhood learning and development. The use of a logic model provided a visual representation of the Kids Together model and its component parts and enabled a theory of change to be inferred, showing how a coordinated and collaborative approached can work across multiple environments.


Can patients with osteoporosis, who should benefit from implementation of the national service framework for older people, be identified from general practice computer records? A pilot study that illustrates the variability of computerized medical records and problems with searching them.
Although UK general practice is highly computerized, comprehensive use of these computers is often limited to registration data and the issue of repeat prescriptions. The recording of diagnostic data is patchy. This study examines whether patients with, or at risk of, osteoporosis can be readily identified from general practice computer records. It reports the findings of a pilot study designed to show the variability of recording the diagnosis of osteoporosis and osteopenia, as well as how useful surrogate markers might be to identify these patients. The study also illustrates the difficulties that even skilled practitioners in a primary care research network experience in extracting clinical data from practice information systems. Computer searches were carried out across six practices in a general practice research network in the south-east of England. Two of these practices had previously undertaken research projects in osteoporosis and were consequently expected to have excellent data quality in osteoporosis. These two practices had a combined list size of 27,500 and the remaining practices had a combined practice population of 43,000 patients. The data were found to be variable with over 10-fold differences between practices in the recorded prevalence of osteoporosis diagnosis as well as its surrogate markers-such as fragility fractures, long-term steroid prescription, etc. There was no difference in data quality between the two practices that had conducted osteoporosis research and the rest of the group, other than in the areas of diagnostic recording and prescribing for osteoporosis and recording of fractures. Issues were raised by the practices that struggled to identify patients at risk of osteoporosis about the limitations of Read classification in this disease area. Practices need further assistance if the patients at risk are to be identified. Without urgent action, it will be difficult for practices to identify the patients who are likely to benefit from Standard 6-'Falls' of the National Service Framework for Older People. These findings also have broader implications as UK general practice moves towards the implementation of a quality-based contract.


First Report of the Occurrence of Cucurbit aphid-borne yellows virus on Oilseed Pumpkin in Serbia.
In July 2008, field-grown oilseed pumpkins (Cucurbita pepo L. 'Olinka') showing severe yellowing and thickening of older leaves were observed in the Kisač locality of Vojvodina Province, Serbia. Symptomatic plants were found only near the borders of the field. Leaf samples collected from 15 symptomatic plants were tested for the presence of four viruses causing the cucurbit yellowing disorder. Total RNAs were extracted from deep frozen plant materials with an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and reverse transcription (RT)-PCR was conducted with the OneStep RT-PCR Kit (Qiagen) following the manufacturer's instructions. RNA extracted from healthy C. pepo and molecular-grade water were included as negative controls in each PCR reaction. Species-specific primers (1,2) failed to detect the presence of three viruses causing the cucurbit yellowing disorder, Cucumber vein yellowing virus, Cucumber yellow stunting disorder virus, and Beet pseudo-yellows virus, in symptomatic samples. When two different sets of CABYV-specific primer pairs, CABYVup/CABYVdown (2) and CE9/CE10 (3), for a 484-bp and a 600-bp fragment of the CP gene of Cucurbit aphid-borne yellows virus (CABYV), respectively, were used for amplification, the former amplified fragments of the expected size from all symptomatic samples, whereas the latter successfully amplified a 600-bp fragment from only 7 of 15 samples. The 600-bp amplified product derived from isolate 145-08 was purified (QIAquick PCR Purification Kit, Qiagen), sequenced in both directions, deposited in GenBank (Accession No. HQ202745), and subjected to sequence analysis by MEGA4 software. Sequence comparisons revealed a high nucleotide identity of 99.8% (100 and 99.5% amino acid identities for the CP and the overlapping MP genes, respectively) with Czech CABYV isolates from C. pepo 'Ovifera' (HM771271-73). A neighbor-joining tree obtained on a 545-bp CP fragment of CABYV isolates available in GenBank database revealed that Serbian CABYV isolate 145-08 was clustered with isolates from Spain, Italy, France, and Tunisia in the Mediterranean subgroup denoted previously (4). In a persistent type transmission test, which was carried out using Aphis gossypii Glover, the aphids were allowed to feed on leaves of the collected sample (145-08) for an acquisition access period of 2 days and then 10 aphids were transferred to each of 20 C. pepo 'Olinka' plants for a 5 day inoculation access period. Transmission was successful in 6 of 20 plants as assessed by the development of a mild yellowing symptom 2 weeks after transmission and confirmed by RT-PCR with the CABYVup/CABYVdown primers. To our knowledge, this is the first record of the occurrence of CABYV in Serbia. The discovery of CABYV on oilseed pumpkin should prompt more detailed surveys and subsequent testing of other cucurbits cultivated in Serbia to establish the distribution and incidence of CABYV in Serbia. References: (1) K. Bananej et al. Plant Dis. 90:1113, 2006. (2) I. N. Boubourakas et al. Plant Pathol. 55:276, 2006. (3) M. Juarez et al. Plant Dis. 88:907, 2004. (4) Q. X. Shang et al. Virus Res. 145:341, 2009.


Experiences of student nurses regarding the bursary system in KwaZulu-Natal province, South Africa.
During 2010, the South African nursing education system was restructured, changing student nurses from having supernumerary status to being bursary holders. Changes with the introduction of this new bursary system included institutional factors and benefits that could be removed from the students, potentially hampering students' sense of belonging. This study aimed to describe the experiences of students receiving bursaries in KwaZulu-Natal (KZN) province and to make recommendations for improving the system to bursary providers, educational institutions and practical settings based on these students' experiences of the bursary system. The experiences of student nurses regarding the bursary system are described within a specified setting comprising two nursing campuses in KZN. A qualitative study design was used and seven focus group interviews were conducted with purposively selected participants, representing the target population of first-, second- and third-year male and female nursing students registered for the Diploma in Nursing (General, Psychiatric, Community) and Midwifery. Two main themes and eight subthemes were identified. The findings indicated that some of the bursary system's experiences were negative as opposed to students having supernumerary status. These experiences had negative socio-economic, psychological, clinical, academic and family impacts. Many concerns related to staff members' attitudes, shortages of nurses and service demands during students' clinical practice assignments. The bursary system was not viewed as being beneficial to students as they did not receive all the benefits from being bursary holders. Support in clinical and academic areas was lacking as they were considered to be employees during their clinical assignments. There is an urgent need to review the bursary system.


Experimental functional response and inter-individual variation in foraging rate of teal (Anas crecca).
The functional response, i.e. the change in per capita food intake rate per time unit with changed food availability, is a widely used tool for understanding the ecology and behaviour of animals. However, waterfowl remain poorly explored in this context. In an aviary experiment we derived a functional response curve for teal (Anas crecca) foraging on rice (Oryza sativa) seeds. We found a linear relationship between intake rate and seed density, as expected for a filter-feeder. At high seed densities we found a threshold, above which intake rate still increased linearly but with a lower slope, possibly reflecting a switch from filter-feeding to a scooping foraging mode. The present study shows that food intake rate in teal is linearly related to food availability within the range of naturally occurring seed densities, a finding with major implications for management and conservation of wetland habitats.


Treatment of antibiotics and antibiotic resistant bacteria in swine wastewater with free chlorine.
Recent recognition of the occurrence of antibiotics in the environment has highlighted concerns regarding potential threats of antibiotics to humans and wildlife. Antibiotics are commonly applied to animals to prevent diseases and promote growth, making livestock agriculture a major source of antibiotic pollution. The purpose of our study was to examine chlorination technology as a method for preventing the release of antibiotics as well as antibiotic-resistant bacteria into the environment from concentrated animal feeding operations. Wastewaters from various sites of two anaerobic lagoon systems, one aerated and the other not, on a swine facility were investigated. Each system consisted of a primary treatment lagoon and a subsequent polishing lagoon. Free chlorine (or monochloramine for comparison) was applied to oxidize antibiotics and to disinfect lagoon bacteria as well. Results indicate that aeration substantially improves lagoon functionality, thereby adding both organic and ammonia removal. Ammonia present in the wastewaters plays a critical role in antibiotics decomposition and bacterial inactivation due to its rapid competition for free chlorine to form monochloramine. Generally, a chlorine dose close to breakpoint is required to achieve complete removal of antibiotics, leading to high consumption of free chlorine in most of the wastewaters examined. However, because of a low ammonia concentration in the polishing lagoon wastewater of the aerated system, a chlorine dose of 100 mg/L can effectively achieve complete removal of both antibiotics and bacteria. On the basis of our experimental findings, a possible strategy for the treatment of swine wastewater is suggested.


Decay-accelerating factor regulates T-cell immunity in the context of inflammation by influencing costimulatory molecule expression on antigen-presenting cells.
Recent studies have indicated a role of complement in regulating T-cell immunity but the mechanism of action of complement in this process remains to be clarified. Here we studied mice deficient in decay-accelerating factor (DAF), a key membrane complement regulator whose deficiency led to increased complement-dependent T-cell immune responses in vivo. By crossing OT-II and OT-I T-cell receptor transgenic mice with DAF-knockout mice, we found that lack of DAF on T cells did not affect their responses to antigen stimulation. Similarly, lack of DAF on antigen-presenting cells (APCs) of naive mice did not alter their T-cell stimulating activity. In contrast, APCs from DAF-knockout mice treated with inflammatory stimuli were found to be more potent T-cell stimulators than cells from similarly treated wild-type mice. Acquisition of higher T-cell stimulating activity by APCs in challenged DAF-knockout mice required C3 and C5aR and was correlated with decreased surface PD-L1 and/or increased CD40 expression. These findings implied that DAF suppressed T-cell immunity as a complement regulator in the context of inflammation but did not play an intrinsic role on T cells or APCs. Collectively, our data suggest a systemic and indirect role of complement in T-cell immunity.


Strategies for bacterial expression of protein-peptide complexes: application to solubilization of papillomavirus E6.
E6 is a small oncoprotein involved in tumorigenesis induced by papillomaviruses (PVs). E6 often recognizes its cellular targets by binding to short motifs presenting the consensus LXXLL. E6 proteins have long resisted structural analysis. We found that bovine papillomavirus type 1 (BPV1) E6 binds the N-terminal LXXLL motif of the cellular protein paxillin with significantly higher affinity as compared to other E6/peptide interactions. Although recombinant BPV1 E6 was poorly soluble in the free state, provision of the paxillin LXXLL peptide during BPV1 E6 biosynthesis greatly enhanced the protein's solubility. Expression of BPV1 E6/LXXLL peptide complexes was carried out in bacteria in the form of triple fusion constructs comprising, from N- to C-terminus, the soluble carrier protein maltose binding protein (MBP), the LXXLL motif and the E6 protein. A TEV protease cleavage site was placed either between MBP and LXXLL motif or between LXXLL motif and E6. These constructs allowed us to produce highly concentrated samples of BPV1 E6, either covalently fused to the C-terminus of the LXXLL motif (intra-molecular complex) or non-covalently bound to it (inter-molecular complex). Heteronuclear NMR measurements were performed and showed that the E6 protein was folded with similar conformations in both covalent and non-covalent complexes. These data open the way to novel structural and functional studies of the BPV1 E6 in complex with its preferential target motif.


Tyrosine phosphorylation of histone H2A by CK2 regulates transcriptional elongation.
Post-translational histone modifications have a critical role in regulating transcription, the cell cycle, DNA replication and DNA damage repair. The identification of new histone modifications critical for transcriptional regulation at initiation, elongation or termination is of particular interest. Here we report a new layer of regulation in transcriptional elongation that is conserved from yeast to mammals. This regulation is based on the phosphorylation of a highly conserved tyrosine residue, Tyr 57, in histone H2A and is mediated by the unsuspected tyrosine kinase activity of casein kinase 2 (CK2). Mutation of Tyr 57 in H2A in yeast or inhibition of CK2 activity impairs transcriptional elongation in yeast as well as in mammalian cells. Genome-wide binding analysis reveals that CK2α, the catalytic subunit of CK2, binds across RNA-polymerase-II-transcribed coding genes and active enhancers. Mutation of Tyr 57 causes a loss of H2B mono-ubiquitination as well as H3K4me3 and H3K79me3, histone marks associated with active transcription. Mechanistically, both CK2 inhibition and the H2A(Y57F) mutation enhance H2B deubiquitination activity of the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex, suggesting a critical role of this phosphorylation in coordinating the activity of the SAGA complex during transcription. Together, these results identify a new component of regulation in transcriptional elongation based on CK2-dependent tyrosine phosphorylation of the globular domain of H2A.


A phase I study to determine the pharmacokinetics and urinary excretion of belinostat and metabolites in patients with advanced solid tumors.
Belinostat is an inhibitor of histone deacetylase enzymes, resulting in DNA repair inhibition and apoptosis. Present data are lacking to provide dosing recommendations in renal insufficiency. The purpose of this trial was to assess the pharmacokinetics (PK) of belinostat and belinostat metabolites in plasma and urine. This was a phase I, single-center, open-label, two-part study. In Part I, patients received single-agent belinostat 1000 mg/m2. Blood and urine samples were collected at pre-specified time points to determine PK of belinostat and metabolites and their elimination in urine. In Part II, patients were permitted to continue belinostat in 21-day cycles on Days 1 through 5 until disease progression, unacceptable toxicity, or according to patient preference. A total of nine patients with advanced solid tumors were treated. Median t max for belinostat was observed 10 min after the start of infusion. Concentrations of belinostat rapidly declined with a t 1/2 of 2.9 h. The mean fraction of belinostat excreted unchanged in urine was 0.926 %. The metabolites belinostat glucuronide and 3-ASBA represented the largest fractions of belinostat dose excreted in urine (30.5 and 4.61 %, respectively), while renal excretion appeared to be a minor route of elimination for the parent belinostat (<1 %). The most common adverse events were nausea, fatigue, and diarrhea. One Grade 3 adverse event (constipation) was thought to be treatment related. Urinary elimination of parent belinostat was minimal, although a combined 36.7 % of belinostat metabolites were excreted in urine. Since these metabolites are primarily inactive, belinostat may not require dosage adjustment in renal dysfunction.


Mechanism of loading the Escherichia coli DNA polymerase III sliding clamp: I. Two distinct activities for individual ATP sites in the gamma complex.
The Escherichia coli DNA polymerase III gamma complex loads the beta clamp onto DNA, and the clamp tethers the core polymerase to DNA to increase the processivity of synthesis. ATP binding and hydrolysis promote conformational changes within the gamma complex that modulate its affinity for the clamp and DNA, allowing it to accomplish the mechanical task of assembling clamps on DNA. This is the first of two reports (Snyder, A. K., Williams, C. R., Johnson, A., O'Donnell, M., and Bloom, L. B. (2004) J. Biol. Chem. 279, 4386-4393) addressing the question of how ATP binding and hydrolysis modulate specific interactions with DNA and beta. Pre-steady-state rates of ATP hydrolysis were slower when reactions were initiated by addition of ATP than when the gamma complex was equilibrated with ATP and were limited by the rate of an intramolecular reaction, possibly ATP-induced conformational changes. Kinetic modeling of assays in which the gamma complex was incubated with ATP for different periods of time prior to adding DNA to trigger hydrolysis suggests a mechanism in which a relatively slow conformational change step (kforward = 6.5 s(-1)) produces a species of the gamma complex that is activated for DNA (and beta) binding. In the absence of beta, 2 of the 3 molecules of ATP are hydrolyzed rapidly prior to releasing DNA, and the 3rd molecule is hydrolyzed slowly. In the presence of beta, all 3 molecules of ATP are hydrolyzed rapidly. These results suggest that hydrolysis of 2 molecules of ATP may be coupled to conformational changes that reduce interactions with DNA, whereas hydrolysis of the 3rd is coupled to changes that result in release of beta.


Laser-induced fluorescence measurement of oxygen atoms above a catalytic combustor surface.
Two-photon laser-induced fluorescence has been used to detect oxygen atoms in the gas phase above a heated, catalytically stabilized combustor. Excitation was at 226 nm and detection at 777 nm. Special care using the collection optics was necessary to avoid detector saturation from intense thermal emission background from the heated plate. The experimental configuration together with considerations of quenching collisions and photochemical production of oxygen atoms are described.


Assessment of risk factors for IgE-mediated sensitization to tetrachlorophthalic anhydride.
Two groups of workers with occupational asthma caused by tetrachlorophthalic anhydride (TCPA) have been reported on; in only one report was specific anti-TCPA antibody demonstrated. We received a request to investigate respiratory problems among workers at a plant where solenoid coils are manufactured with the use of epoxy resin and where a new TCPA hardener had been introduced in 1988. In response, we conducted a cross-sectional evaluation of 52 current workers and assayed serum samples from 49 workers for TCPA-human serum albumin-specific IgE and IgG by ELISA. Mean airborne TCPA concentrations ranged from 0.21 to 0.39 mg/m3 in 1989 and May 1990 but were considerably lower (less than 0.009 to 0.13 mg/m3) by July 1990 when ventilation was introduced. Workers were classified in four departments as molders (most directly working with TCPA), those intermittently exposed to TCPA, coil assemblers, and office workers. There was a high prevalence of reported work-related respiratory symptoms (from 27% to 39%). The mean decline in FEV1 over the shift in the molders (down 4.1%) was significantly different from the change in the office group (increased 2.5%). Intrashift change in FEV1 was inversely correlated with log personal TCPA concentration (r = -0.47). Of 49 current workers, 15 (31%) had elevated serum TCPA-human serum albumin-specific IgE antibody levels and 19 (39%) had elevated specific IgG levels. The prevalence of specific IgE was highest in the mold and intermittent groups (54%), in comparison with the coil assembly (25%) and office (0%) groups. Those workers who were IgE positive were significantly less likely to be current smokers and had worked significantly longer since 1988. Of those hired in 1988 or after, the proportion of workers who were IgE positive increased with duration of employment category (p value for trend 0.022). Since ventilation was installed and TCPA exposures reduced to less than 0.1 mg/m3, there has been a marked decrease in symptoms and no new cases of occupational asthma among newly hired workers at the plant.


Formal reasoning on qualitative models of coinfection of HIV and Tuberculosis and HAART therapy.
Several diseases, many of which nowadays pandemic, consist of multifactorial pathologies. Paradigmatic examples come from the immune response to pathogens, in which cases the effects of different infections combine together, yielding complex mutual feedback, often a positive one that boosts infection progression in a scenario that can easily become lethal. HIV is one such infection, which weakens the immune system favouring the insurgence of opportunistic infections, amongst which Tuberculosis (TB). The treatment with antiretroviral therapies has shown effective in reducing mortality. An in-depth understanding of complex systems, like the one consisting of HIV, TB and related therapies, is an open great challenge, on the boundaries of bioinformatics, computational and systems biology. We present a simplified formalisation of the highly dynamic system consisting of HIV, TB and related therapies, at the cellular level. The progression of the disease (AIDS) depends hence on interactions between viruses, cells, chemokines, the high mutation rate of viruses, the immune response of individuals and the interaction between drugs and infection dynamics. We first discuss a deterministic model of dual infection (HIV and TB) which is able to capture the long-term dynamics of CD4 T cells, viruses and Tumour Necrosis Factor (TNF). We contrast this model with a stochastic approach which captures intrinsic fluctuations of the biological processes. Furthermore, we also integrate automated reasoning techniques, i.e. probabilistic model checking, in our formal analysis. Beyond numerical simulations, model checking allows general properties (effectiveness of anti-HIV therapies) to be verified against the models by means of an automated procedure. Our work stresses the growing importance and flexibility of model checking techniques in bioinformatics. In this paper we i) describe HIV as a complex case of infectious diseases; ii) provide a number of different formal descriptions that suitably account for aspects of interests; iii) suggest that the integration of different models together with automated reasoning techniques can improve the understanding of infections and therapies through formal analysis methodologies. We argue that the described methodology suitably supports the study of viral infections in a formal, automated and expressive manner. We envisage a long-term contribution of this kind of approaches to clinical Bioinformatics and Translational Medicine.


Perfluorochemicals and human semen quality: the LIFE study.
The relation between persistent environmental chemicals and semen quality is evolving, although limited data exist for men recruited from general populations. We examined the relation between perfluorinated chemicals (PFCs) and semen quality among 501 male partners of couples planning pregnancy. Using population-based sampling strategies, we recruited 501 couples discontinuing contraception from two U.S. geographic regions from 2005 through 2009. Baseline interviews and anthropometric assessments were conducted, followed by blood collection for the quantification of seven serum PFCs (perfluorosulfonates, perfluorocarboxylates, and perfluorosulfonamides) using tandem mass spectrometry. Men collected a baseline semen sample and another approximately 1 month later. Semen samples were shipped with freezer packs, and analyses were performed on the day after collection. We used linear regression to estimate the difference in each semen parameter associated with a one unit increase in the natural log-transformed PFC concentration after adjusting for confounders and modeling repeated semen samples. Sensitivity analyses included optimal Box-Cox transformation of semen quality end points. Six PFCs [2-(N-methyl-perfluorooctane sulfonamido) acetate (Me-PFOSA-AcOH), perfluorodecanoate (PFDeA), perfluorononanoate (PFNA), perfluorooctane sulfonamide (PFOSA), perfluorooctane sulfonate (PFOS), and perfluorooctanoic acid (PFOA)] were associated with 17 semen quality end points before Box-Cox transformation. PFOSA was associated with smaller sperm head area and perimeter, a lower percentage of DNA stainability, and a higher percentage of bicephalic and immature sperm. PFDeA, PFNA, PFOA, and PFOS were associated with a lower percentage of sperm with coiled tails. Select PFCs were associated with certain semen end points, with the most significant associations observed for PFOSA but with results in varying directions.


A multi-measure examination of interoception in people with recent nonsuicidal self-injury.
Self-injurious behaviors (SIB) are highly dangerous, yet prediction remains weak. Novel SIB correlates must be identified, such as impaired interoception. This study examined whether two forms of interoceptive processing (accuracy and sensibility) for multiple sensations (general, cardiac, and pain) differed between people with and without recent nonsuicidal self-injury (NSSI). Participants were adults with recent (n = 48) NSSI and with no history of SIBs (n = 55). Interoceptive sensibility was assessed with self-reports. Interoceptive accuracy for cardiac sensations was assessed using the heartbeat tracking task. Interoceptive accuracy for pain was assessed with a novel metric that mirrored the heartbeat tracking test. Participants with recent NSSI reported significantly lower interoceptive sensibility for general sensations relative to people without SIBs. Groups did not differ on interoceptive sensibility for cardiac sensations or pain. Groups also did not differ on interoceptive accuracy for cardiac sensations. The NSSI group exhibited significantly lower interoceptive accuracy for pain compared with the No SIB group. Interoceptive impairment in people with NSSI may vary by interoceptive domain and sensation type. Diminished interoceptive accuracy for sensations relevant to the pathophysiology of self-injury may be a novel SIB correlate.


A comparative study of different Leishmania tropica isolates from Iran: correlation between infectivity and cytochemical properties.
Five isolates of Leishmania tropica from southwest Iran were studied to identify correlates among human disease, animal infectivity, and surface biochemistry. Clinical patterns of the disease in humans differed. One striking strain, LT-249, produced a small dry lesion which did not heal during four years of observation. Infectivity of these L. tropica for mice was correlated with lectin agglutination patterns and interaction with macrophages. There was also a significant difference among the five isolates regarding infectivity for BALB/c mice; isolate LT-249 was not infective whereas all the others were. All isolates agglutinated with Concanavalin A (Con A), Ricinus communis and soybean agglutinin but not with four other lectins listed. However, Leishmania isolate LT-249 showed much poorer agglutination with all lectins than did the other four isolates. Two isolates were selected for detailed study of attachment to macrophages, one, LT-249, which was not infective and one, LT-252, which was infective for BALB/c mice. The number of promastigotes which attached to macrophages in vitro was the same, but the mechanism of attachment differed since only the LT-252 bound predominantly by Con A-mannose receptor interaction. These results indicate a correlation among animal infectivity, lectin agglutination, and promastigote-macrophage attachment. In particular one isolate of L. tropica which caused prolonged infection in humans was not infective in BALB/c mice, showed poor agglutination with lectins, and bound to macrophages by a different mechanism than did other isolates from the same region.


Genetic susceptibility to non-insulin dependent diabetes mellitus and glucose intolerance are located in HLA region.
To test the hypothesis that the genetic susceptibility to non-insulin dependent diabetes mellitus is the same as that to insulin dependent disease and to see whether glucose intolerance is associated with specific HLA haplotypes. Population based study of men in 1989 first tested for glucose tolerance in 1984. HLA haplotypes, including HLA-A, C, B, DR, and DQ, were defined serologically. HLA haplotype data from a population based Finnish study of childhood diabetes were used for predicting non-insulin dependent diabetes and impaired glucose tolerance. Two communities in Finland. Representative cohort of Finnish men aged 70-89, comprising 98 men with non-insulin dependent diabetes mellitus and a randomly selected group of 74 men, who served as controls, who were tested for glucose tolerance twice within five years. Non-insulin dependent diabetes, impaired glucose tolerance, blood glucose concentration. Diabetes associated HLA haplotypes were present in 94% (85/90) of diabetic subjects, 79% (27/34) of subjects with impaired glucose tolerance, and only 13% (3/23) of non-diabetic subjects. In this group of elderly men sensitivity of the diabetes associated HLA haplotypes for non-insulin dependent diabetes and impaired glucose tolerance was 90%, specificity 87%, and predictive power 97%. Mean fasting blood glucose concentration was only just significantly higher in men with diabetes associated haplotypes than in men with no such haplotypes, but there was a substantial difference in blood glucose values two hours after glucose loading (10.4 and 6.4 mmol/l in men with diabetes associated HLA haplotypes and men with no such haplotypes, respectively (p < 0.0001)). These findings support the hypothesis that specific HLA haplotypes exhibit a common genetic determinant for insulin dependent and non-insulin dependent diabetes. Furthermore, HLA is a major genetic determinant of glucose intolerance in elderly Finnish men. The belief that the HLA predisposition to diabetes is specific for insulin dependent diabetes mellitus is largely incorrect.


Plasmonics Enhanced Smartphone Fluorescence Microscopy.
Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.


Circulating MicroRNAs Highly Correlate to Expression of Cartilage Genes Potentially Reflecting OA Susceptibility-Towards Identification of Applicable Early OA Biomarkers.
To identify and validate circulating micro RNAs (miRNAs) that mark gene expression changes in articular cartilage early in osteoarthritis (OA) pathophysiology process. Within the ongoing RAAK study, human preserved OA cartilage and plasma (N = 22 paired samples) was collected for RNA sequencing (respectively mRNA and miRNA). Spearman correlation was determined for 114 cartilage genes consistently and significantly differentially expressed early in osteoarthritis and 384 plasma miRNAs. Subsequently, the minimal number of circulating miRNAs serving to discriminate between progressors and non-progressors was assessed by regression analysis and area under receiver operating curves (AUC) was calculated with progression data and plasma miRNA sequencing from the GARP study (N = 71). We identified strong correlations (ρ ≥ |0.7|) among expression levels of 34 unique plasma miRNAs and 21 genes, including 4 genes that correlated with multiple miRNAs. The strongest correlation was between let-7d-5p and EGFLAM (ρ = -0.75, P = 6.9 × 10-5). Regression analysis of the 34 miRNAs resulted in a set of 7 miRNAs that, when applied to the GARP study, demonstrated clinically relevant predictive value with AUC > 0.8 for OA progression over 2 years and near-clinical value for progression over 5 years- (AUC = 0.8). We show that plasma miRNAs levels reflect gene expression levels in cartilage and can be exploited to represent ongoing pathophysiological processes in articular cartilage. We advocate that identified signature of 7 plasma miRNAs can contribute to direct further studies toward early biomarkers predictive for progression of osteoarthritis over 2 and 5 years.


Programmable mechanical metamaterials: the role of geometry.
We experimentally and numerically study the role of geometry for the mechanics of biholar metamaterials, which are quasi-2D slabs of rubber patterned by circular holes of two alternating sizes. We recently showed how the response to uniaxial compression of these metamaterials can be programmed by lateral confinement. In particular, there is a range of confining strains εx for which the resistance to compression becomes non-trivial-non-monotonic or hysteretic-in a range of compressive strains εy. Here we show how the dimensionless geometrical parameters t and χ, which characterize the wall thickness and size ratio of the holes that pattern these metamaterials, can significantly tune these ranges over a wide range. We study the behavior for the limiting cases where the wall thickness t and the size ratio χ become large, and discuss the new physics that arises there. Away from these extreme limits, the variation of the strain ranges of interest is smooth with porosity, but the variation with size ratio evidences a cross-over at low χ from biholar to monoholar (equal sized holes) behavior, related to the elastic instabilities in purely monoholar metamaterials. Our study provides precise guidelines for the rational design of programmable biholar metamaterials, tailored to specific applications, and indicates that the widest range of programmability arises for moderate values of both t and χ.


Systemic ropivacaine diminishes pain sensitization processes: a randomized, double-blinded, placebo-controlled, crossover study in healthy volunteers.
Ropivacaine is a local anesthetic widely used for regional anesthesia. One of its advantages is low toxicity at plasma concentrations reached systemically during continuous peripheral or central nervous block. The objective of this study was to test the effect of systemic ropivacaine on pain, hyperalgesia, dynamic allodynia, and flare response. This randomized, double-blinded, placebo-controlled, crossover study was carried out in at the Clinical Trials Centre, University of Zurich, Switzerland. Twenty healthy male volunteers were included in the study. Exclusion criteria were contraindications or hypersensitivity to local anesthetics, vulnerable subjects (intellectually or mental impaired), drug, alcohol or nicotine abuse, known peripheral neuropathies, diabetes mellitus and/or congestive heart disease. Ropivacaine and saline were infused intravenously during a subcutaneous electrical stimulation. The stimulation software adjusted the stimulus strength according to the rating on a numeric rating scale (NRS; 0-10) maintaining a NRS of 5. Areas of punctate hyperalgesia, dynamic allodynia, and flare response were measured before and after the infusion. The area of hyperalgesia increased significantly with saline (303 ± 380%, P < 0.05) and ropivacaine (186 ± 137%, P < 0.05). The area of allodynia (253 ± 299%, P < 0.05) and flare response (112 ± 24%, P < 0.05) increased only during the placebo infusion. The results of this study imply that systemic ropivacaine may diminish pain sensitization processes.


Lost and found: recovery of the holotype of the ocellated angelshark, Squatina tergocellatoides Chen, 1963 (Squatinidae), with comments on western Pacific squatinids.
The ocellated angelshark, Squatina tergocellatoides, Chen, 1963 is redescribed from the holotype, which was thought to be lost. Its recent recovery has allowed for a revised description, including new data, and comparison to other Western Pacific squatinids. Squatina tergocellatoides can be distinguished from its congeners by three pairs of prominent large black spots, each with a diameter greater than eye length; two on each pectoral fin at anterior and posterior angles and one on each side near the tail base; another three pairs of lesser defined spots, one large spot on base of each dorsal fin and one located laterally on each side of tail located below first dorsal fin. Ventral surface is uniformly white to cream coloured, and margins of pectoral fins and tail similar in colour to dorsal side. Pectoral fins with angular lateral apices and rounded posterior lobe, pelvic fin tips not reaching origin of first dorsal fin, strongly fringed nasal barbels, small inter-orbital space, head and mouth lengths, broad internarial width and pelvic fin base, a very small pelvic girdle width, and a caudal fin with triangular ventral lobe greater in length than dorsal lobe. Comments on additional specimens are provided, as well as observations on biogeography. A review of western Pacific squatinids is also provided.


Mechanism of loading the Escherichia coli DNA polymerase III sliding clamp: II. Uncoupling the beta and DNA binding activities of the gamma complex.
Sliding clamps tether DNA polymerases to DNA to increase the processivity of synthesis. The Escherichia coli gamma complex loads the beta sliding clamp onto DNA in an ATP-dependent reaction in which ATP binding and hydrolysis modulate the affinity of the gamma complex for beta and DNA. This is the second of two reports (Williams, C. R., Snyder, A. K., Kuzmic, P., O'Donnell, M., and Bloom, L. B. (2004) J. Biol. Chem. 279, 4376-4385) addressing the question of how ATP binding and hydrolysis regulate specific interactions with DNA and beta. Mutations were made to an Arg residue in a conserved SRC motif in the delta' and gamma subunits that interacts with the ATP site of the neighboring gamma subunit. Mutation of the delta' subunit reduced the ATP-dependent beta binding activity, whereas mutation of the gamma subunits reduced the DNA binding activity of the gamma complex. The gamma complex containing the delta' mutation gave a pre-steady-state burst of ATP hydrolysis, but at a reduced rate and amplitude relative to the wild-type gamma complex. A pre-steady-state burst of ATP hydrolysis was not observed for the complex containing the gamma mutations, consistent with the reduced DNA binding activity of this complex. The differential effects of these mutations suggest that ATP binding at the gamma1 site may be coupled to conformational changes that largely modulate interactions with beta, whereas ATP binding at the gamma2 and/or gamma3 site may be coupled to conformational changes that have a major role in interactions with DNA. Additionally, these results show that the "arginine fingers" play a structural role in facilitating the formation of a conformation that has high affinity for beta and DNA.


Maternal Haploid, a Metalloprotease Enriched at the Largest Satellite Repeat and Essential for Genome Integrity in Drosophila Embryos.
The incorporation of the paternal genome into the zygote during fertilization requires chromatin remodeling. The maternal haploid (mh) mutation in Drosophila affects this process and leads to the formation of haploid embryos without the paternal genome. mh encodes the Drosophila homolog of SPRTN, a conserved protease essential for resolving DNA-protein cross-linked products. Here we characterize the role of MH in genome maintenance. It is not understood how MH protects the paternal genome during fertilization, particularly in light of our finding that MH is present in both parental pronuclei during zygote formation. We showed that maternal chromosomes in mh mutant embryos experience instabilities in the absence of the paternal genome, which suggests that MH is generally required for chromosome stability during embryogenesis. This is consistent with our finding that MH is abundantly present on chromatin throughout the cell cycle. Remarkably, MH is prominently enriched at the 359-bp satellite repeats during interphase, which becomes unstable without MH. This dynamic localization and specific enrichment of MH at the 359 repeats resemble that of Topoisomerase 2 (Top2), suggesting that MH regulates Top2, possibly as a protease for the resolution of Top2-DNA intermediates. We propose that maternal MH removes proteins specifically enriched on sperm chromatin. In the absence of that function, paternal chromosomes are precipitously lost. This mode of paternal chromatin remodeling is likely conserved and the unique phenotype of the Drosophila mh mutants represents a rare opportunity to gain insights into the process that has been difficult to study.


four-jointed is required for intermediate growth in the proximal-distal axis in Drosophila.
Genes capable of translating positional information into regulated growth lie at the heart of morphogenesis, yet few genes with this function have been identified. Mutants in the Drosophila four-jointed (fj) gene show reduced growth and altered differentiation only within restricted sectors of the proximal-distal (PD) axis in the leg and wing, thus fj is a candidate for a gene with this coordination function. Consistent with a position-sensitive role, we show that fj is expressed in a regional pattern in the developing leg, wing, eye and optic lobe. The fj gene encodes a novel type II membrane glycoprotein. When the cDNA is translated in an in vitro translation system in the presence of exogenous microsomal membranes, the intralumenal portion of some of the molecules is cleaved, yielding a secreted C-terminal fragment. We propose that fj encodes a secreted signal that functions as a positive regulator of regional growth and differentiation along the PD axis of the imaginal discs.


Rapid complex mode decomposition of vector beams by common path interferometry.
We have used common path interferometry for rapid determination of the electric field and complex modal content of vector beams, which have spatially-varying polarization. We combine a reference beam with a signal beam prior to a polarization beam splitter for stable interferograms that preserve intermodal phase shifts even in noisy environments. Interferometric decomposition into optical modes (IDIOM) provides a direct, sensitive measure of the complete electric field, enabling rapid modal decomposition and is ideally suited to single-frequency laser sources. We apply the technique to beams exiting optical fibers that support up to 10 modes. We also use the technique to characterize the fibers by determining a scattering matrix that transforms an input superposition of modes into an output superposition. Furthermore, because interferograms are linear in the field, this technique is very sensitive and can accurately reconstruct beams with signal-to-noise << 1.


Na-K-ATPase regulates intercellular communication in the vascular wall via cSrc kinase-dependent connexin43 phosphorylation.
Communication between vascular smooth muscle cells (VSMCs) is dependent on gap junctions and is regulated by the Na-K-ATPase. The Na-K-ATPase is therefore important for synchronized VSMC oscillatory activity, i.e., vasomotion. The signaling between the Na-K-ATPase and gap junctions is unknown. We tested here the hypothesis that this signaling involves cSrc kinase. Intercellular communication was assessed by membrane capacitance measurements of electrically coupled VSMCs. Vasomotion in isometric myograph, input resistance, and synchronized [Ca2+]i transients were used as readout for intercellular coupling in rat mesenteric small arteries in vitro. Phosphorylation of cSrc kinase and connexin43 (Cx43) were semiquantified by Western blotting. Micromole concentration of ouabain reduced the amplitude of norepinephrine-induced vasomotion and desynchronized Ca2+ transients in VSMC in the arterial wall. Ouabain also increased input resistance in the arterial wall. These effects of ouabain were antagonized by inhibition of tyrosine phosphorylation with genistein, PP2, and by an inhibitor of the Na-K-ATPase-dependent cSrc activation, pNaKtide. Moreover, inhibition of cSrc phosphorylation increased vasomotion amplitude and decreased the resistance between cells in the vascular wall. Ouabain inhibited the electrical coupling between A7r5 cells, but pNaKtide restored the electrical coupling. Ouabain increased cSrc autophosphorylation of tyrosine 418 (Y418) required for full catalytic activity whereas pNaKtide antagonized it. This cSrc activation was associated with Cx43 phosphorylation of tyrosine 265 (Y265). Our findings demonstrate that Na-K-ATPase regulates intercellular communication in the vascular wall via cSrc-dependent Cx43 tyrosine phosphorylation.


Insight into subtilisin E-S7 cleavage pattern based on crystal structure and hydrolysates peptide analysis.
The X-ray crystallographic structure of the mature form of subtilisin E-S7 (SES7) at 1.90 Å resolution is reported here. Structural comparisons between the previously reported propeptide-subtilisin E complex (1SCJ) and our mature form subtilisin E-S7 (6O44) provide insight into active site adjustments involved in catalysis and specificity. To further investigate the protease substrate selectivity mechanism, we used SES7 to hydrolyze skim milk and analyzed the hydrolysates by LC-MS for peptide identification. The cleavage pattern suggests a high preference for proline at substrate P2 position. The results based on the peptide analysis are consistent with our structural observations, which is instrumental in future protein engineering by rational design. Furthermore, the ACE-inhibitor and NLN-inhibitor activity of the hydrolysates were determined to assess the utility of SES7 for further industrial applications; IC50-ACE = 67 ± 0.92 μg/mL and IC50-NLN = 263 ± 13 μg/mL.


A meaningful MESS (Medical Education Scholarship Support).
Graduate medical education faculty bear the responsibility of demonstrating active research and scholarship; however, faculty who choose education-focused careers may face unique obstacles related to the lack of promotion tracks, funding, career options, and research opportunities. Our objective was to address education research and scholarship barriers by providing a collaborative peer-mentoring environment and improve the production of research and scholarly outputs. We describe a Medical Education Scholarship Support (MESS) group created in 2013. MESS is an interprofessional, multidisciplinary peer-mentoring education research community that now spans multiple institutions. This group meets monthly to address education research and scholarship challenges. Through this process, we develop new knowledge, research, and scholarly products, in addition to meaningful collaborations. MESS originated with eight founding members, all of whom still actively participate. MESS has proven to be a sustainable unfunded local community of practice, encouraging faculty to pursue health professions education (HPE) careers and fostering scholarship. We have met our original objectives that involved maintaining 100% participant retention; developing increased knowledge in at least seven content areas; and contributing to the development of 13 peer-reviewed publications, eight professional presentations, one Masters of Education project, and one educational curriculum. The number of individuals engaged in HPE research continues to rise. The MESS model could be adapted for use at other institutions, thereby reducing barriers HPE researchers face, providing an effective framework for trainees interested in education-focused careers, and having a broader impact on the education research landscape.