An analysis of patients transferred to a tertiary oncological intensive care unit for defined procedures.
Up to 50 000 intensive care unit interhospital transfers occur annually in the United States. To determine the prevalence, characteristics, and outcomes of cancer patients transferred from an intensive care unit in one hospital to another intensive care unit at an oncological center and to evaluate whether interventions planned before transfer were performed. Data on transfers for planned interventions from January 2008 through December 2012 were identified retrospectively. Demographic and clinical variables, receipt of planned interventions, and outcome data were analyzed. Of 4625 admissions to an intensive care unit at the oncological center, 143 (3%) were transfers from intensive care units of other hospitals. Of these, 47 (33%) were transfers for planned interventions. Patients' mean age was 57 years, and 68% were men. At the time of intensive care unit transfer, 20 (43%) were receiving mechanical ventilation. Interventions included management of airway (n = 19) or gastrointestinal (n = 2) obstruction, treatment of tumor bleeding (n = 12), chemotherapy (n = 10), and other (n = 4). A total of 37 patients (79%) received the planned interventions within 48 hours of intensive care unit arrival; 10 (21%) did not because their signs and symptoms abated. Median intensive care unit and hospital lengths of stay at the oncological center were 4 and 13 days, respectively. Intensive care unit and hospital mortality rates were 11% and 19%, respectively. Deaths occurred only in patients who received interventions. Interhospital transfers of cancer patients to an intensive care unit at an oncological center are infrequent but are most commonly done for direct interventional care. Most patients received planned interventions soon after transfer.


Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide.
The thermal stabilities of RNA:DNA hybrids are substantially greater than those of DNA:DNA duplexes in aqueous electrolyte solutions containing high concentrations of formamide. Association rates to form DNA:DNA duplexes and DNA:RNA hybrids have been measured in these solvents. There is a temperature range in which DNA:DNA rates are negligible and RNA:DNA rates close to optimal.


Motivation in deliberate self-harm.
Fifty adult patients presenting with deliberate self-harm at the Royal Victoria Hospital were given a choice of nine reasons for their actions. Most chose more than one reason and all but two of the 24 who said that they wished to die chose at least one other motive. There were no trends with respect to sex, past history, or method of deliberate self-harm. These results illustrate the complexity of this condition and show the importance of investigating motives beyond simply the intent to die.


Spermicide used alone for contraception.
Spermicides have been used as contraceptives for thousands of years. Despite this long use, only recently have studies examined the comparative efficacy and acceptability of these vaginal medications. Spermicides contain an active ingredient (most commonly nonoxynol-9) and a formulation used to disperse the product, such as foam or vaginal suppository. This review examined all known randomized controlled trials of a spermicide used alone for contraception. In August 2013, we searched the following computerized databases for randomized controlled trials of spermicides for contraception: CENTRAL, MEDLINE, POPLINE, LILACS, EMBASE, ClinicalTrials.gov, and ICTRP. For the initial review, we examined the reference lists of trials found as well as those of review articles and textbook chapters. We included any trial of a commercial product used alone for contraception. Each included trial must have provided sufficient information to determine pregnancy rates. Two authors independently extracted information from the trials identified. We did not conduct a meta-analysis, since most trials had large losses to follow up. We entered the data into tables and presented the results descriptively. We located reports from 14 trials for the initial review, but have not identified any new trials since then. In the largest trial to date, the gel (Advantage S) containing the lowest dose of nonoxynol-9 (52.5 mg) was significantly less effective in preventing pregnancy than were gels with higher doses of the same agent (100 mg and 150 mg). Probabilities of pregnancy by six months were 22% for the 52.5 mg gel, 16% for the 100 mg dose, and 14% for the 150 mg dose. In the same trial, the three different vehicles with 100 mg of nonoxynol-9 had similar efficacy. Interpretation of these figures is limited, since 39% of participants discontinued the method or were lost from the trial. Few important differences in efficacy emerged in other trials. The probability of pregnancy varied widely in reported trials. A gel containing nonoxynol-9 52.5 mg was inferior to two other products tested in the largest trial. Aside from this finding, personal characteristics and behavior of users may be more important than characteristics of the spermicide products in determining the probability of pregnancy. Gel was liked more than the film or vaginal suppository in the largest trial. Spermicide trials have the dual challenges of difficult recruitment and high discontinuation rates; the latter threatens trial validity.


Scar size and characteristics assessed by CMR predict ventricular arrhythmias in ischaemic cardiomyopathy: comparison of previously validated models.
Sudden cardiac death is a major cause of mortality in patients with ischaemic cardiomyopathy. Risk stratification remains challenging. Currently, there is growing interest in scar characteristic assessment as a predictor of sudden cardiac death using cardiac magnetic resonance imaging (CMR). Standard analysis methods are lacking. The present study evaluated previously validated methods of scar assessment by CMR with late gadolinium enhancement (LGE) in their ability to predict ventricular tachyarrhythmias. Patients with ischaemic cardiomyopathy who received an implantable cardioverter defibrillator for primary prevention and in whom a LGE-CMR was performed, were included. Scar core size, peri-infarct zone and total scar size, which is defined as the sum of the core size and peri-infarct zone, were assessed using three previously validated models, and their ability to predict ventricular tachyarrhythmias was evaluated. Fifty-five patients were included (mean age 64.6 ± 10.8 years, 43 men). During a median follow-up of 2.0 years (IQR 1.0-3.0 years) 26% of patients reached the endpoint of ventricular tachyarrhythmia. All scar characteristics (ie, total scar size, scar core size and peri-infarct zone) of the three methods were predictors of the endpoint (p < 0.01). Total scar size was comparable, whereas scar core size and peri-infarct zone varied significantly between the tested models. Receiver operating characteristic curves of the different scar characteristics showed comparable areas under the curve varying from 0.721 to 0.812. LGE-CMR-derived scar tissue characteristics are of predictive value for the occurrence of ventricular tachyarrhythmias in patients with ischaemic cardiomyopathy. Additional estimation of scar core size and/or peri-infarct zone does not appear to increase the diagnostic accuracy over total scar size alone.


Quantitative analyses and modelling to support achievement of the 2020 goals for nine neglected tropical diseases.
Quantitative analysis and mathematical models are useful tools in informing strategies to control or eliminate disease. Currently, there is an urgent need to develop these tools to inform policy to achieve the 2020 goals for neglected tropical diseases (NTDs). In this paper we give an overview of a collection of novel model-based analyses which aim to address key questions on the dynamics of transmission and control of nine NTDs: Chagas disease, visceral leishmaniasis, human African trypanosomiasis, leprosy, soil-transmitted helminths, schistosomiasis, lymphatic filariasis, onchocerciasis and trachoma. Several common themes resonate throughout these analyses, including: the importance of epidemiological setting on the success of interventions; targeting groups who are at highest risk of infection or re-infection; and reaching populations who are not accessing interventions and may act as a reservoir for infection,. The results also highlight the challenge of maintaining elimination 'as a public health problem' when true elimination is not reached. The models elucidate the factors that may be contributing most to persistence of disease and discuss the requirements for eventually achieving true elimination, if that is possible. Overall this collection presents new analyses to inform current control initiatives. These papers form a base from which further development of the models and more rigorous validation against a variety of datasets can help to give more detailed advice. At the moment, the models' predictions are being considered as the world prepares for a final push towards control or elimination of neglected tropical diseases by 2020.


Stress relaxation of the respiratory system in developing piglets.
To characterize the effect of postnatal development on the viscoelastic behavior of the respiratory system, we quantified the amplitude and time course of stress relaxation in the lungs and chest wall of seven newborn and eight 8-wk-old anesthetized piglets. Stress relaxation was distinguished from other dissipative pressure losses by performing airway occlusions at various constant inspiratory flows and fitting the pressure decays that ensue during the occlusions to a double-exponential function. We found that the amplitude of stress relaxation related linearly to the increase in elastic recoil (and, by extension, in the volume) of the lungs, chest wall, and respiratory system during the inflations preceding the occlusions. On the average, the slope of this relationship was 38-44% lower in the 8-wk-old than in the newborn piglets for the lungs and was not different for the chest wall. The time course of stress relaxation, expressed as a time constant, was not influenced by age. Our results indicate that respiratory system viscoelasticity is sensitive to the geometric and structural changes experienced by the lungs during the period of rapid somatic growth that follow birth in most mammals.


Albumin Exchange in Alzheimer's Disease: Might CSF Be an Alternative Route to Plasma?
Amyloid β (Aβ) in brain parenchyma is thought to play a central role in the pathogenesis of Alzheimer's disease (AD). Aβ is transported from the brain to the plasma via complex transport mechanisms at the blood-brain barrier (BBB). About 90-95% of plasma Aβ may be bound to albumin. Replacement of serum albumin in plasma has been proposed as a promising therapy for AD. However, the efficacy of this approach may be compromised by altered BBB Aβ receptors in AD, as well as multiple pools of Aβ from other organs in exchange with plasma Aβ, competing for albumin binding sites. The flow of interstitial fluid (ISF) into cerebrospinal fluid (CSF) is another major route of Aβ clearance. Though the concentration of albumin in CSF is much lower than in plasma, the mixing of CSF with ISF is not impeded by a highly selective barrier and, hence, Aβ in the two pools is in more direct exchange. Furthermore, unlike in plasma, Aβ in CSF is not in direct exchange with multiple organ sources of Aβ. Here we consider albumin replacement in CSF as an alternative method for therapeutic brain Aβ removal and describe the possible advantages and rationale supporting this hypothesis.


Seeing a Mycobacterium-Infected Cell in Nanoscale 3D: Correlative Imaging by Light Microscopy and FIB/SEM Tomography.
Mycobacteria pose a threat to the world health today, with pathogenic and opportunistic bacteria causing tuberculosis and non-tuberculous disease in large parts of the population. Much is still unknown about the interplay between bacteria and host during infection and disease, and more research is needed to meet the challenge of drug resistance and inefficient vaccines. This work establishes a reliable and reproducible method for performing correlative imaging of human macrophages infected with mycobacteria at an ultra-high resolution and in 3D. Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) tomography is applied, together with confocal fluorescence microscopy for localization of appropriately infected cells. The method is based on an Aclar poly(chloro-tri-fluoro)ethylene substrate, micropatterned into an advantageous geometry by a simple thermomoulding process. The platform increases the throughput and quality of FIB/SEM tomography analyses, and was successfully applied to detail the intracellular environment of a whole mycobacterium-infected macrophage in 3D.


Toxicity of indium-111 on the radiolabeled lymphocyte.
The radiolabeling of lymphocytes with 111In has resulted in detectable toxic changes in the cells. The mechanisms of toxicity for lymphocytes have been related to the label's radioactivity and to the chelator used to mediate the intracellular localization. These mechanisms were examined by assessing cellular function with mitogen-mediated blastogenesis after labeling lymphocytes with either the chelator (tropolone) alone, 111In complexed with tropolone, or cadmium (the decay product of 111In) complexed with tropolone. Successful lymphocyte labeling with 111In was shown to be dependent upon the concentration of the chelator (tropolone). Increasing concentrations of tropolone inhibited lymphocyte function to a variable degree. Further reduction in cellular function was detected after incorporation of a constant amount of 111In or 111In's decay product, cadmium. Lymphocyte function was decreased by these two labels in a parallel linear manner. This same toxic effect was seen after labeling with small constant amounts of tropolone and increasing quantities of 111In or cadmium. Thus, although both the required chelator and the radiobiologic exposure have a deleterious effect on the lymphocyte, significant lymphocyte toxicity appears to result from the metal-to-cell interaction as a result of the metal decay product (cadmium).


The next generation of synthetic cannabinoids: Detection, activity, and potential toxicity of pent-4en and but-3en analogues including MDMB-4en-PINACA.
A new class of synthetic cannabinoids has emerged as new psychoactive substances (NPS). Similar in structure to JWH-022, these substances contain alkene modifications to the tail region of the synthetic cannabinoid core structure, and nomenclature denotes these new analogues as pent-4en or but-3en species. Internationally, two analogues from this new series recently emerged: MDMB-4en-PINACA and MMB-4en-PICA. Previously, data regarding activity and potential toxicity were not available. In vitro assessment of cannabinoid receptor 1 (CB1) activation via the β-arrestin 2 recruitment was studied for three (3) pent-4en analogues, one (1) but-3en analogue, and one (1) principal metabolite. MDMB-4en-PINACA (2.47 nM, 239%), MDMB-4en-PICA (11.5 nM, 302%), and MDMB-3en-BINACA (14.3 nM, 286%) were highly potent and efficacious (comparison: JWH-018, 25.3 nM, 100%), while the potencies of MMB-4en-PICA and MDMB-4en-PINACA 3,3-dimethylbutanoic acid were markedly lower. Modifications to core and tail structural features (i.e., indole vs. indazole) led to relatively small differences in potency, while changes among the head region led to larger differences. Sample-mining and data-mining conducted on toxicology samples led to the identification of MDMB-4en-PINACA in 25 forensic toxicology cases, including postmortem and impaired driving investigations, with case details and limited histories described herein. Moderate geographical distribution of MDMB-4en-PINACA was noted in the United States with emergence in the Northeast, Midwest, South, and West regions. Results from toxicology testing paired with case history show the potential for MDMB-4en-PINACA to cause or contribute to impairment or death. Forensic scientists, public health and public safety officials, law enforcement, clinicians, medical examiners, and coroners should consider involvement of emergent synthetic cannabinoids in their work and that new analogues containing an alkene tail can retain similar or increased potency and toxicity.


[Regression of general anesthesia in patients with cerebral palsy: a comparative study using the bispectral index.].
It has been said that patients with cerebral palsy (CP) present a higher sensitivity to anesthetic agents, which may affect adversely the awakening from anesthesia. The objective of this study was to evaluate the recovery of patients with CP compared with patients without CNS disease. The study population was composed of children ages 5 to 15, divided in two groups: those with a diagnosis of cerebral palsy and those without CNS disease. All of them underwent general anesthesia with sevoflurane associated with nitrous oxide and 50% oxygen. Besides the parameters commonly monitored, they also underwent EEG-BIS monitoring. Regression of general anesthesia during awakening from anesthesia, tracheal extubation, and the immediate postanesthetic period were evaluated according to the criteria adopted by Saraiva. The speed of elimination of the anesthetic agents was also evaluated, correlating it with the EEG-BIS values, and comparing the two groups of patients. Statistical analysis was done through exploratory analysis of the data and statistical testing comparing the means. The statistical difference was considered significant for values of p smaller or equal to 5%. Fifty-six patients were evaluated: 31 with CP and 25 without CNS disease. Both groups were homogenous regarding age and gender. Baseline EEG-BIS values for CP patients were smaller than those for the control group, and this difference was statistically significant (p = 0.04). During the first five minutes after discontinuation of sevoflurane, the elevation of the EEG-BIS values for CP patients was slower than for the control group. The speed of elimination of the anesthetic agent was similar in both groups. At 15 minutes, 48% of the patients in the control group were in stage II of anesthetic regression versus 29% of the patients with cerebral palsy. At 30 minutes, 72% of the patients in the control group were in condition to be discharged, in stage IV of anesthetic regression, versus 41.9% of the patients with CP. Patients with cerebral palsy presented a slower return to the conscious state when compared with patients in the control group.


[Childhood amnesia and emotional trauma. Easiest to prompt the smallest children to provide erroneous details concerning abuse].
A basic understanding of children's memories for emotional trauma is of importance for physicians and other health care professionals. The aim of this paper is to describe recent research and discuss important dilemmas within this field. The ability of infants and toddlers to verbally describe autobiographical memories is for several reasons limited. Very early experiences of e.g. intense pain can leave traces in the nervous system but of a kind which is not specific enough to allow retrospective conclusions about the nature of these experiences. Research during the past decades has relatively consistently demonstrated that pre-school children are more vulnerable to suggestion including abuse-related suggestions than are older children and adults. However, many children who have been subject to sexual abuse seem to maintain a lifelong silence about their experiences. The use of autobiographical testimony from young children is more complex and risky than many professionals believed about a decade ago. Establishing the knowledge derived from recent years' research in this area as a basis for clinical practice is one of the most important tasks for the future.


Thermodynamics of writhe in DNA minicircles from molecular dynamics simulations.
DNA supercoiling plays a role in genetic control by imposing torsional stress. This can induce writhe, which changes the global shape of the DNA. We have used atomistic molecular dynamics simulations to partition the free energy changes driving the writhing and unwrithing transitions in supercoiled minicircle DNA. The calculations show that while writhing is energetically driven, the unwrithing transition occurs because the circular state has a higher configurational entropy than the plectoneme. Writhing improves the van der Waals interactions between stacked bases, but can be suppressed by electrostatic repulsion within the negatively charged backbone strands in low salt conditions where electrostatic screening is poor. The free energy difference between circular and plectonemic DNA is determined by such a delicate balance of opposing thermodynamic terms that any perturbation in the environment, such as a change in salt concentration, can be sufficient to convert between these two states. This switchable behavior provides a mechanism for supercoiled DNA to store and communicate biological information physically as well as chemically.


Insulin antibodies are associated with lipoatrophy but also with lipohypertrophy in children and adolescents with type 1 diabetes.
To evaluate clinical and immunological factors that are associated with lipodystrophy, i.e. lipoatrophy and lipohypertrophy, in diabetic children and adolescents. We investigated in a cross-sectional study 112 children and adolescents (age 1.1-19.1 yrs.) with type 1 diabetes. To grade lipodystrophy, we developed a clinical score ranging from normal (grade 0), moderate hypertrophy of subcutaneous tissue (grade 1), severe hypertrophy with increased density of tissue (grade 2) to lipoatrophy (grade 3). In all children, grade of lipodystrophy, antibodies against insulin (IA) or beta cell antigens (IA-2 and GAD) and clinical parameters were documented. The antibodies against insulin (IA) increased significantly after diabetes manifestation and initiation of insulin treatment, while beta cell specific antibodies (IA-2, GAD) did not. Lipoatrophy (grade 3) was seen in 4 children, severe lipohypertrophy (grade 2) in 18 and moderate lipohypertrophy (grade 1) in 27 children. No alteration of injection sites was found in 63 children. Amongst clinical and immunological parameters, IA levels were significantly associated with hypertrophy or atrophy of injection sites. The strong association of lipoatrophy and lipohypertrophy with insulin antibodies might suggest that autoimmune phenomena with insulin play a role in the development of both. Despite an association of IA and lipodystrophy in type 1 diabetic children, the causal link between the two remains unproven and requires further longitudinal exploration.


Predictors of outcome in civilian gunshot wounds to the head.
Civilian gunshot wounds to the head (GSWH) are often deadly, but some patients with open cranial wounds need medical and surgical management and are potentially good candidates for acceptable functional recovery. The authors analyzed predictors of favorable clinical outcome (Glasgow Outcome Scale scores of 4 and 5) after GSWH over a 24-month period. The authors posited 2 questions: First, what percentage of civilians with GSWH died in the state of Maryland in a given period of time? Second, what were the predictors of favorable outcome after GSWH? The authors examined demographic, clinical, imaging, and acute care data for 786 civilians who sustained GSWH. Univariate and logistic regression analyses were used to analyze the data. Of the 786 patients in this series, 712 (91%) died and 74 (9%) completed acute care in 9 trauma centers. Of the 69 patients admitted to one Maryland center, 46 (67%) eventually died. In 48 patients who were resuscitated, the Injury Severity Score was 26.2, Glasgow Coma Scale (GCS) score was 7.8, and an abnormal pupillary response (APR) to light was present in 41% of patients. Computed tomography indicated midline shift in 17%, obliteration of basal cisterns in 41.3%, intracranial hematomas in 34.8%, and intraventricular hemorrhage in 49% of cases. When analyzed for trajectory, 57.5% of bullet slugs crossed midcoronal, midsagittal, or both planes. Two subsets of admissions were studied: 27 patients (65%) who had poor outcome (25 patients who died and 2 who had severe disability) and 15 patients (35%) who had a favorable outcome when followed for a mean period of 40.6 months. Six patients were lost to follow-up. Univariate analysis indicated that admission GCS score (p < 0.001), missile trajectory (p < 0.001), surgery (p < 0.001), APR to light (p = 0.002), patency of basal cisterns (p = 0.01), age (p = 0.01), and intraventricular bleed (p = 0.03) had a significant relationship to outcome. Multivariable logistic regression analysis indicated that GCS score and patency of the basal cistern were significant determinants of outcome. Exclusion of GCS score from the regression models indicated missile trajectory and APR to light were significant in determining outcome. Admission GCS score, trajectory of the missile track, APR to light, and patency of basal cisterns were significant determinants of outcome in civilian GSWH.


Functional properties of dopaminergic neurones in the mouse olfactory bulb.
The olfactory bulb of mammals contains a large population of dopaminergic interneurones within the glomerular layer. Dopamine has been shown both in vivo and in vitro to modulate several aspects of olfactory information processing, but the functional properties of dopaminergic neurones have never been described due to the inability to recognize these cells in living preparations. To overcome this difficulty, we used a transgenic mouse strain harbouring an eGFP (enhanced green fluorescent protein) reporter construct under the promoter of tyrosine hydroxylase, the rate-limiting enzyme for cathecolamine synthesis. As a result, we were able to identify dopaminergic neurones (TH-GFP cells) in living preparations and, for the first time, we could study the functional properties of such neurones in the olfactory bulb, in both slices and dissociated cells. The most prominent feature of these cells was the autorhythmicity. In these cells we identified five main voltage-dependent conductances: the two having largest amplitude were a fast transient Na(+) current and a delayed rectifier K(+) current. In addition, we observed three smaller inward currents, sustained by Na(+) ions (persistent type) and by Ca(2)(+) ions (LVA and HVA). Using pharmacological tools and ion substitution methods we showed that the pacemaking process is supported by the interplay of the persistent Na(+) current and of a T-type Ca(2)(+) current. We carried out a complete kinetical analysis of the five conductances present in these cells, and developed a Hodgkin-Huxley model of TH-GFP cells, capable of reproducing accurately the properties of living cells, including autorhytmicity, and allowing a precise understanding of the process.


Effects of kidney transplantation on cognitive performance in a pediatric population.
The cognitive functioning of 20 children and adolescents with end-stage renal disease was assessed 1 to 3 weeks prior to the onset of hemodialysis, and at 1 month and 1 year after successful kidney transplantation, and results were compared with those of a matched control group. A battery of intelligence, achievement, problem solving, learning, memory, and attention tasks were administered. Both groups had significantly improved scores over time on most measures. The group with renal disease exhibited significantly greater improvement than the control group from initial testing to 1 month after transplantation on the performance IQ and full-scale IQ as well as in mathematics achievement. This significant difference was not maintained, however, at 1 year after transplantation. Cognitive performance was less impaired the later the onset of renal failure or the fewer the years in end-stage renal disease. BUN nitrogen, serum creatinine levels, and BP did not consistently correlate with any of the cognitive or academic achievement measures.


Dose prediction for repurposing nitazoxanide in SARS-CoV-2 treatment or chemoprophylaxis.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been declared a global pandemic by the World Health Organisation and urgent treatment and prevention strategies are needed. Many clinical trials have been initiated with existing medications, but assessments of the expected plasma and lung exposures at the selected doses have not featured in the prioritisation process. Although no antiviral data is currently available for the major phenolic circulating metabolite of nitazoxanide (known as tizoxanide), the parent ester drug has been shown to exhibit in vitro activity against SARS-CoV-2. Nitazoxanide is an anthelmintic drug and its metabolite tizoxanide has been described to have broad antiviral activity against influenza and other coronaviruses. The present study used physiologically-based pharmacokinetic (PBPK) modelling to inform optimal doses of nitazoxanide capable of maintaining plasma and lung tizoxanide exposures above the reported nitazoxanide 90% effective concentration (EC 90 ) against SARS-CoV-2. A whole-body PBPK model was constructed for oral administration of nitazoxanide and validated against available tizoxanide pharmacokinetic data for healthy individuals receiving single doses between 500 mg SARS-CoV-2 4000 mg with and without food. Additional validation against multiple-dose pharmacokinetic data when given with food was conducted. The validated model was then used to predict alternative doses expected to maintain tizoxanide plasma and lung concentrations over the reported nitazoxanide EC 90 in >90% of the simulated population. Optimal design software PopDes was used to estimate an optimal sparse sampling strategy for future clinical trials. The PBPK model was validated with AAFE values between 1.01 SARS-CoV-2 1.58 and a difference less than 2-fold between observed and simulated values for all the reported clinical doses. The model predicted optimal doses of 1200 mg QID, 1600 mg TID, 2900 mg BID in the fasted state and 700 mg QID, 900 mg TID and 1400 mg BID when given with food, to provide tizoxanide plasma and lung concentrations over the reported in vitro EC 90 of nitazoxanide against SARS-CoV-2. For BID regimens an optimal sparse sampling strategy of 0.25, 1, 3 and 12h post dose was estimated. The PBPK model predicted that it was possible to achieve plasma and lung tizoxanide concentrations, using proven safe doses of nitazoxanide, that exceed the EC 90 for SARS-CoV-2. The PBPK model describing tizoxanide plasma pharmacokinetics after oral administration of nitazoxanide was successfully validated against clinical data. This dose prediction assumes that the tizoxanide metabolite has activity against SARS-CoV-2 similar to that reported for nitazoxanide, as has been reported for other viruses. The model and the reported dosing strategies provide a rational basis for the design (optimising plasma and lung exposures) of future clinical trials of nitazoxanide in the treatment or prevention of SARS-CoV-2 infection.


Joint association of physical activity and body weight with subsequent physical and mental functioning: a follow-up study.
Physical inactivity and overweight are major threats to public health. However, it is not well understood to what extent physical activity might counteract the harmful effects of overweight on functioning. Thus, we examined the joint associations of leisure-time physical activity and body mass index (BMI) with subsequent physical and mental functioning over a follow-up of five to seven years. The data were derived from the Helsinki Health Study, which is a cohort study among employees of the City of Helsinki, Finland. The baseline postal survey data were collected among 40-60-year-old employees in 2000-02 (n = 8960, response rate 67%), and the follow-up data in 2007 among all baseline survey respondents (n = 7332, response rate 83%). We divided the participants into six groups according to their amount of physical activity (inactive, moderately active and highly active) and their relative weight (normal weight and overweight). Highly active normal-weight participants were used as a reference group in all the analyses. Poor functioning was defined as the lowest quartile of the Short Form 36 (SF-36) health survey's physical and mental component summaries, with the follow-up cut-off point also applied at baseline. We used logistic regression analysis adjusted for age, gender, baseline functioning, smoking, alcohol use, marital status, socioeconomic position and working conditions. At baseline 48% of the participants were overweight and 11% were inactive. After adjustments inactivity was associated with poor physical functioning at follow-up both among the normal-weight (OR 1.51, 95% CI 1.09-2.10) and overweight (OR 2.02, 95% CI 1.56-2.63) groups. Being overweight regardless of activity level was associated with poor physical functioning. Poor physical functioning was practically equally common among the highly active overweight group and the inactive normal-weight group. After adjustments, for mental functioning, only inactivity among the overweight was associated with poor mental functioning (OR 1.39, 95% CI 1.08-1.80). Physical activity is likely to be beneficial for physical and mental functioning among both those with overweight and normal weight. However, maintaining normal weight is also important for good physical functioning. Therefore, efforts should be made to recommend people to engage in physical activity regardless of weight.


Capsaicin activates a nonselective cation channel in cultured neonatal rat dorsal root ganglion neurons.
Capsaicin (CAP), a neurotoxin, has been reported to activate a nonselective cation current in dorsal root ganglion (DRG) neurons. In this paper, we identify and describe the properties of CAP-activated single channels in cultured neonatal rat DRG neurons. We first identified CAP-sensitive whole-cell currents that reversed near 0 mV in physiological solution. In solution containing 140 mM Na+, extracellular application of CAP to outside-out patches caused activation of an ion channel in a concentration-dependent manner (EC50 = 1.1 microM). The channel was blocked by the CAP antagonist capsazepine (10 microM). The channel was also activated by 2-10 nM resiniferatoxin, a potent analog of CAP. In symmetrical 140 mM Na+, the single-channel slope conductances were 45.3 +/- 1.0 and 80.0 +/- 4.2 pS at -60 and +60 mV, respectively, showing outward rectification (n = 9). The reversal potential did not shift significantly when Na+ was replaced by K+, Cs+, Rb+, or Li+, showing that the channel discriminated poorly among cations. The channel was also permeable to Ca2+. Although acid (pH < 6.2) was suggested to be an endogenous activator of the CAP receptor, an acid solution (pH 5.9-6.0) failed to activate the channels in outside-out patches. This is the first clear demonstration of the presence of the CAP-activated ion channel in DRG neuron. Opening of these ligand-gated, cation-selective channels gives rise to the whole-cell CAP-activated current in DRG neurons and may underlie the neurotoxic effects of CAP.


Characterization of Drosophila fruitless-gal4 transgenes reveals expression in male-specific fruitless neurons and innervation of male reproductive structures.
The fruitless (fru) gene acts in the central nervous system (CNS) of Drosophila melanogaster to establish male sexual behavior. Genetic dissection of the locus has shown that one of the fru gene's promoter, P1, controls the spatial and temporal expression of male-specific FruM proteins critical to determining stereotypical male sexual behavior. By using the Gal4-expression system, we show that a 16-kb fragment of the fru P1 promoter's 5' regulatory region drives the expression of Gal4 in a subset of FruM-expressing neurons within both the pupal and adult CNS. Colocalization of FruM and a Gal4-responsive reporter shows that the fru(P1)-gal4 fusion construct generates expression in both previously characterized FruM-expressing neurons as well as within cells of both the CNS and the peripheral nervous system that have not been demonstrated as FruM-expressing. Gal4-expressing neurons are shown to innervate abdominal organs directly relevant to fru function; specifically, the muscle of Lawrence (MOL) and the male internal reproductive organs. Innervations of the latter are shown to originate from identified FruM-serotonergic neurons. Furthermore, we show that the MOL neuromuscular junction is sexually dimorphic. Finally, we describe Gal4 expression in neurites innervating male reproductive structures that are hypothesized to be targets of fru function. Isolation of the regulatory sequences controlling the expression of fru in the CNS, therefore, provides a potent tool for the manipulation of FruM-expressing neurons and for understanding the cellular basis of Drosophila reproductive behavior.


Transfusion rate using intravenous tranexamic acid in hip revision surgery.
Hip revision surgery is associated with a high probability of the necessity for a blood transfusion. Different studies support the use of tranexamic acid (TXA) to decrease the rate of transfusions in primary hip surgery. Nevertheless, the use of this drug in hip revision surgery has not yet been widely accepted. The aim of our study was to establish the rate of blood transfusion with and without TXA in hip revision surgery. We retrospectively studied 125 hip revision surgery patients operated on between 2011 and 2014. We divided our series into 2 groups: the TXA group with 61 patients (in which a 1000 mg dose of TXA was used before the skin incision and a second identical dose after skin closure) and a control group with 64 patients. We analysed the red blood cell (RBC) transfusion rates and their odds risk as well as the presence of collateral complications. Average RBC transfusion was 2.7 units/patient (range 0-6) in the control group compared to 1.6 units/patient (range 0-6) in the TXA group. A 90.11% (odds ratio [OR] 0.098; confidence interval [CI] 0.02-0.04; p<0.0029) odds risk reduction for transfusion of at least 1 unit of erythrocyte blood cell was observed in the TXA group. Complications associated with the TXA were similar in both groups. The benefits of TXA have been shown in elective hip replacement. In this study, TXA proved to be safe and efficacious in reducing the need for transfusions following revision total hip arthroplasty.


Antileukemic Effect of Tualang Honey on Acute and Chronic Leukemia Cell Lines.
Complementary medicine using natural product as antitumor is on the rise. Much research has been performed on Tualang Honey and it was shown to have therapeutic potential in wound healing, and antimicrobial activity and be antiproliferative against several cancer models such as human osteosarcoma (HOS), human breast (MCF-7 and MDA-MB-231), and cervical (HeLa) cancer cell lines. To date, there was limited study on antileukemic properties of Tualang (Koompassia excelsa) Honey. The aim of this study was to evaluate the antileukemic effect of Tualang Honey on acute and chronic leukemia cell lines. Leukemia cell lines (K562 and MV4-11) and human mononuclear cell isolated from peripheral blood were grown in RPM1 1640 culture medium. The cells were incubated with increasing concentrations of Tualang Honey. After incubation, the evaluation of viability and apoptosis was performed. The morphological changes of leukemia cells were the presence of cytoplasmic blebs followed by apoptotic bodies and round shape of cells. IC50 against K562 and MV4-11 was determined. Tualang Honey gave 53.9% and 50.6% apoptosis activity on K562 and MV4-11, respectively, while on human mononuclear cell it was 37.4%. Tualang Honey has the apoptosis-inducing ability for acute and chronic myeloid leukemia (K562 and MV4-11) cell lines.


Resveratrol-Mediated Repression and Reversion of Prostatic Myofibroblast Phenoconversion.
Resveratrol, a phytoalexin found in berries, peanuts, grapes, and red wine, inhibits oxidation, inflammation, and cell proliferation and collagen synthesis in multiple cell types and or animal models. It represses collagen deposition in the vasculature, heart, lung, kidney, liver, and esophagus in animal models and may have some utility as an anti-fibrotic. Recent studies have shown that increased collagen deposition and tissue stiffness in the peri-urethral area of the prostate are associated with lower urinary tract dysfunction (LUTD) and urinary obstructive symptoms. The aim of this study was to determine whether Resveratrol might be useful to inhibit or revert TGFβ- and/or CXCL12-mediated myofibroblast phenoconversion of prostate fibroblasts in vitro, and therefore whether the use of anti-fibrotic therapeutics might be efficacious for the treatment of LUTD. Primary prostate and lung tissues were explanted and fibroblast monolayers expanded in vitro. Primary and N1 immortalized prostate stromal fibroblasts, as well as primary fibroblasts cultured from a normal lung and one affected by idiopathic pulmonary fibrosis (IPF) for comparison, were grown in serum-free defined media supplemented with vehicle, TGFβ or CXCL12, pre- or post-treatment with Resveratrol, and were evaluated using immunofluorescence for alpha smooth muscle actin (αSMA) and collagen I (COL1) protein expression and assessed for cell proliferation, apoptosis, and COL1 and EGR1 transcript expression. This study showed that low concentrations of Resveratrol (≤50 μM) had no effect on N1 or primary prostate fibroblast cell proliferation, apoptosis, or COL1 or EGR1 gene transcription but repressed and reversed myofibroblast phenoconversion. As expected, these same effects were observed for IPF lung fibroblasts though higher levels of Resveratrol (≥100uM) were required. Taken together, these data suggest that, like lung fibroblasts, prostate fibroblast to myofibroblast phenoconversion can be both repressed and reversed by Resveratrol treatment. Thus, anti-fibrotic therapeutics might be efficacious for the treatment of LUTD.


Risk factors for third-generation cephalosporin resistant Enterobacteriaceae in gestational urine cultures: A retrospective cohort study based on centralized electronic health records.
Third-generation-cephalosporin resistant Enterobacteriaceae (3GCR-EB) carriage in pregnant women poses challenges for infection control and therapeutic decisions. The factors associated with multidrug resistant Enterobacteriaceae carriage in the gestational period are not well documented. The aim of our study was to identify risk factors associated with 3GCR-EB isolation in gestational urine cultures. The study was designed as retrospective cohort based on centralized electronic health records database. Women delivered in Clalit Health Services hospitals in Israel in 2009-2013 and provided urine culture(s) during pregnancy were included. Multivariable analysis using the Generalized Estimating Equations model was used to assess risk factors for 3GCR-EB isolation in gestational urine cultures. The study included 15,282 pregnant women with urine cultures yielding Enterobacteriaceae (EB). The proportion of 3GCR-EB in EB isolates was 3.9% (n = 603). The following risk factors were associated with 3GCR-EB isolation: multiple hospital admissions during the year before delivery (OR,1.47;95% CI,1.21-1.79), assisted fertilization procedure (OR,1.53; 95% CI,1.12-2.10), Arab ethnicity (OR,1.22; 95% CI,1.03-1.45), multiple antibiotic courses (OR,1.76; 95% CI,1.29-2.40), specifically, cephalosporins (OR,1.56; 95% CI,1.26-1.95), fluoroquinolones (OR,1.34; 95% CI,1.04-1.74), or nitrofurantoin (OR,1.29; 95% CI,1.02-1.64). The risk factors identified by this study for 3GCR-EB in gestation, can be easily generalized for pregnant women in the Israeli population. Moreover, these risk factors, other than ethnicity, are applicable to pregnant women worldwide. The information of previous antibiotic treatments, hospitalization in the last year and assisted fertilization procedure can be easily accessed and used for appropriate infection control practices and antimicrobial therapy.


Alert-QSAR. Implications for electrophilic theory of chemical carcinogenesis.
Given the modeling and predictive abilities of quantitative structure activity relationships (QSARs) for genotoxic carcinogens or mutagens that directly affect DNA, the present research investigates structural alert (SA) intermediate-predicted correlations A(SA) of electrophilic molecular structures with observed carcinogenic potencies in rats (observed activity, A = Log[1/TD(50)], i.e., [Formula: see text]). The present method includes calculation of the recently developed residual correlation of the structural alert models, i.e., [Formula: see text]. We propose a specific electrophilic ligand-receptor mechanism that combines electronegativity with chemical hardness-associated frontier principles, equality of ligand-reagent electronegativities and ligand maximum chemical hardness for highly diverse toxic molecules against specific receptors in rats. The observed carcinogenic activity is influenced by the induced SA-mutagenic intermediate effect, alongside Hansch indices such as hydrophobicity (LogP), polarizability (POL) and total energy (Etot), which account for molecular membrane diffusion, ionic deformation, and stericity, respectively. A possible QSAR mechanistic interpretation of mutagenicity as the first step in genotoxic carcinogenesis development is discussed using the structural alert chemoinformation and in full accordance with the Organization for Economic Co-operation and Development QSAR guidance principles.


Improved learning in a large-enrollment physics class.
We compared the amounts of learning achieved using two different instructional approaches under controlled conditions. We measured the learning of a specific set of topics and objectives when taught by 3 hours of traditional lecture given by an experienced highly rated instructor and 3 hours of instruction given by a trained but inexperienced instructor using instruction based on research in cognitive psychology and physics education. The comparison was made between two large sections (N = 267 and N = 271) of an introductory undergraduate physics course. We found increased student attendance, higher engagement, and more than twice the learning in the section taught using research-based instruction.


A large-scale neurocomputational model of task-oriented behavior selection and working memory in prefrontal cortex.
The prefrontal cortex (PFC) is crucially involved in the executive component of working memory, representation of task state, and behavior selection. This article presents a large-scale computational model of the PFC and associated brain regions designed to investigate the mechanisms by which working memory and task state interact to select adaptive behaviors from a behavioral repertoire. The model consists of multiple brain regions containing neuronal populations with realistic physiological and anatomical properties, including extrastriate visual cortical regions, the inferotemporal cortex, the PFC, the striatum, and midbrain dopamine (DA) neurons. The onset of a delayed match-to-sample or delayed nonmatch-to-sample task triggers tonic DA release in the PFC causing a switch into a persistent, stimulus-insensitive dynamic state that promotes the maintenance of stimulus representations within prefrontal networks. Other modeled prefrontal and striatal units select cognitive acceptance or rejection behaviors according to which task is active and whether prefrontal working memory representations match the current stimulus. Working memory task performance and memory fields of prefrontal delay units are degraded by extreme elevation or depletion of tonic DA levels. Analyses of cellular and synaptic activity suggest that hyponormal DA levels result in increased prefrontal activation, whereas hypernormal DA levels lead to decreased activation. Our simulation results suggest a range of predictions for behavioral, single-cell, and neuroimaging response data under the proposed task set and under manipulations of DA concentration.


The Role of Culture in Maintaining Post-Partum Sexual Abstinence of Swazi Women.
Eswatini is one of the countries in the African continent where post-partum sexual abstinence is practiced. Beside scarcity of research exploring sexual abstinence in Eswatini, there are only a few studies that explore post-partum abstinence across HIV-positive and negative women in sub-Saharan Africa. The study explored the practice of post-partum sexual abstinence in Swazi women and examined how cultural beliefs influence and promotes the perpetuation of the practice. The study population consisted of post-partum women who were selected, using purposive sampling. Thematic approach was used for data analysis. Despite feeling that the period for post-partum, sexual abstinence was long; the participants adhered to the practice as prescribed by their culture. Nevertheless, they felt that the practice is imposed on women only because while they are observing post-partum abstinence, their partners get to sleep with other sexual partners. They raised concerns that the practice increases the risk of acquiring HIV and sexually transmitted infections. There is an element of coercion to the practice of post-partum abstinence, the myths and misconceptions around the early resumption of sexual intercourse forces the practice on women. At the family and community level, the discussions to change the way sexual abstinence is viewed and practiced are crucial.


Cardiovascular disease and osteoporosis: is there a link between lipids and bone?
Atherosclerotic heart disease and osteoporosis are both diseases of old age. Evidence is accumulating for a link between vascular and bone disease. Calcification is a common feature of atherosclerotic plaques, and osteoporosis is associated with both atherosclerosis and vascular calcification. However, the relationship of vascular calcification to the pathogenesis of atherosclerosis remains incompletely understood. Hormone replacement therapy has beneficial effects in the prevention of both atherosclerosis and osteoporosis. Bisphosphonates inhibit bone resorption and are used in the treatment of osteoporosis, whereas the statins inhibit cholesterol biosynthesis and are used for the treatment of atherosclerosis. We have reviewed recent advances in the knowledge of the actions of bisphosphonates and statins at the cellular, molecular and end-organ levels in order to examine the relationship between cardiovascular disease and osteoporosis and to explore the link between lipids and bones. These studies suggest that the mechanism of actions of these two classes of drugs at the cellular level may not be mutually exclusive. There are some early clinical data to complement these findings, suggesting that statins increase bone density and bisphosphonates may have a beneficial effect in vivo on plasma lipid levels and on the atherosclerotic process. Properly designed prospective studies that examine the effect of statins on bone density and fractures, as well as the effects of bisphosphonates on lipid profiles, atherosclerotic progression and cardiovascular morbidity and mortality are needed to define clearly the clinical effects and potential new roles for these agents.


First entry into nonmetal-centred porphycenes: synthesis of a phosphorus(v) complex of octaethylporphycene.
A phosphorus(v) complex of porphycene [P(OEPc)(OMe)2]PF6 (OEPc = 2,3,6,7,12,13,16,17-octaethylporphycenato dianion) has been synthesized and structurally characterized as the first porphycene derivative incorporating nonmetal elements in the macrocyclic cavity. An extremely low energy level of the LUMO is observed, which is rationalized by the low-lying π* orbital of the porphycene ring coupled with the insertion of the highly-inductive pentavalent phosphorus centre. The compound is luminescent in bright red with a quantum yield of 22.7% in CH2Cl2 at 298 K.


Where Do Early Career Researchers Stand on Open Science Practices? A Survey Within the Max Planck Society.
Open science (OS) is of paramount importance for the improvement of science worldwide and across research fields. Recent years have witnessed a transition toward open and transparent scientific practices, but there is still a long way to go. Early career researchers (ECRs) are of crucial relevance in the process of steering toward the standardization of OS practices, as they will become the future decision makers of the institutional change that necessarily accompanies this transition. Thus, it is imperative to gain insight into where ECRs stand on OS practices. Under this premise, the Open Science group of the Max Planck PhDnet designed and conducted an online survey to assess the stance toward OS practices of doctoral candidates from the Max Planck Society. As one of the leading scientific institutions for basic research worldwide, the Max Planck Society provides a considerable population of researchers from multiple scientific fields, englobed into three sections: biomedical sciences, chemistry, physics and technology, and human and social sciences. From an approximate total population of 5,100 doctoral candidates affiliated with the Max Planck Society, the survey collected responses from 568 doctoral candidates. The survey assessed self-reported knowledge, attitudes, and implementation of different OS practices, namely, open access publications, open data, preregistrations, registered reports, and replication studies. ECRs seemed to hold a generally positive view toward these different practices and to be interested in learning more about them. Furthermore, we found that ECRs' knowledge and positive attitudes predicted the extent to which they implemented these OS practices, although levels of implementation were rather low in the past. We observed differences and similarities between scientific sections. We discuss these differences in terms of need and feasibility to apply these OS practices in specific scientific fields, but additionally in relation to the incentive systems that shape scientific communities. Lastly, we discuss the implications that these results can have for the training and career advancement of ECRs, and ultimately, for the consolidation of OS practices.


First Report of Impatiens necrotic spot virus on Begonia in Bosnia and Herzegovina.
Impatiens necrotic spot virus (INSV) and Tomato spotted wilt virus (TSWV) are the most serious viral pathogens in the production of ornamental plants in Europe and North America (1). During a survey for the presence of tospoviruses in July 2012, potted begonia hybrids (Begonia × tuberhybrida Voss) exhibiting foliar chlorotic rings and zonal spots accompanied by leaf necrosis and distortion, were observed in a greenhouse in the vicinity of Banja Luka (Bosnia and Herzegovina). Leaf samples collected from 12 symptomatic plants were analyzed for the presence of INSV and TSWV by commercial double-antibody sandwich (DAS)-ELISA kits (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls and extracts from healthy begonia leaves were included in each ELISA. INSV was detected serologically in all 12 begonia samples and all tested samples were negative for TSWV. Five healthy plants of each Petunia × hybrida and Nicotiana benthamiana were mechanically inoculated with sap from an ELISA-positive sample (157-12) using chilled 0.01 M phosphate buffer (pH 7) containing 0.1% sodium sulphite. Local necrotic lesions on P. × hybrida and systemic chlorotic mottling on N. benthamiana were observed on all inoculated plants 4 and 10 days post-inoculation, respectively. For further confirmation of INSV infection, total RNAs were extracted from all ELISA-positive begonia plants as well as mechanically inoculated N. benthamiana plants with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and used as template in reverse transcription (RT)-PCR. RT-PCR was performed with the OneStep RT-PCR Kit (Qiagen) using primer pair INSV-589 and TOS-R15 (3), specific to the partial INSV nucleocapsid (N) gene. Total RNA obtained from Serbian INSV isolate from a begonia (GenBank Accession No. HQ724289) and RNA extracts from healthy begonia plants were used as positive and negative controls, respectively. All naturally and mechanically infected plants as well as the positive control yielded an amplicon of the expected size (589 bp), while no amplification products were obtained from the healthy controls. The RT-PCR product derived from the isolate 157-12 was sequenced directly after purification with QIAquick PCR Purification Kit (Qiagen) and submitted to GenBank (KC494869). Pairwise comparison of the 157-12 isolate N sequence with other homologous sequences available in GenBank, conducted using MEGA5 software (2), revealed that begonia isolate from Bosnia and Herzegovina showed the highest nucleotide identity of 99.7% (100% amino acid identity) with the Chinese INSV isolate (FN400772) originating from Oncidium sp. To our knowledge, this is the first report of INSV on begonia in Bosnia and Herzegovina. Begonias are very popular and widely grown ornamentals in Bosnia and Herzegovina and the presence of a new and devastating pathogen could represent a serious threat for its production. Since begonia is commonly grown together with numerous ornamental plants susceptible to INSV, further investigations are needed in order to prevent spread of this potentially harmful pathogen to new hosts in Bosnia and Herzegovina. References: (1) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (2) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011. (3) H. Uga and S. Tsuda. Phytopathology 95:166, 2005.


Macroscopic Rotator Cuff Tendinopathy and Histopathology Do Not Predict Repair Outcomes of Rotator Cuff Tears.
Numerous studies have identified factors that may affect the chances of rotator cuff healing after surgery. Intraoperative tendon quality may be used to predict healing and to determine type of repair and/or consideration of augmentation. There are no data that correlate how gross tendon morphology and degree of tendinopathy affect patient outcome or postoperative tendon healing. Purpose/Hypothesis: The purposes of this study were to (1) compare the gross appearance of the tendon edge during arthroscopic rotator cuff repair with its histological degree of tendinopathy and (2) determine if gross appearance correlated with postoperative repair integrity. The hypothesis was that gross (macroscopic) tendon with normal thickness, no delamination, and elastic tissue before repair would have a correlation with low Bonar scores, higher postoperative American Shoulder and Elbow Surgeons (ASES) scores, and increased rates of postoperative tendon healing on ultrasound. Cross-sectional study; Level of evidence, 3. A total of 105 patients undergoing repair of medium-size (1-3 cm) full-thickness rotator cuff tears were enrolled in the study. Intraoperatively, the supraspinatus tendon was rated on thickness, fraying, and stiffness. Tendon tissue was recovered for histological analysis based on the Bonar scoring system. Postoperative ASES and ultrasound assessment of healing were obtained 1 year after repair. Correlation between gross appearance of the tendon and rotator cuff histology was determined. Of the 105 patients, 85 were followed the study to completion. The mean age of the patients was 61.6 years; Bonar score, 7.5; preoperative ASES score, 49; and postoperative ASES score, 86. Ninety-one percent of repairs were intact on ultrasound. Gross appearance of torn rotator cuff tendon tissue did not correlate with histological appearance. Neither histological (Bonar) score nor gross appearance correlated with multivariate analysis of ASES score, postoperative repair status, or demographic data. The degree of tendinopathy did not correlate with morphological appearance of the tendon. Neither of these parameters correlated with healing or patient outcome. This study suggests that the degree of tendinopathy, unlike muscle atrophy, may not be predictive of outcomes and that, on appearance, poor quality tendon has adequate healing capacity. Therefore, abnormal gross tendon appearance should not affect the repair effort or technique.


Association Between Executive Function and Problematic Adolescent Driving.
Motor vehicle collisions (MVCs) are one of the leading causes of injury and death for adolescents. Driving is a complex activity that is highly reliant on executive function (EF) to safely navigate through the environment. Little research has examined the efficacy of using self-reported EF measures for assessing adolescent driving risk. This study examined the Behavior Rating Inventory of Executive Function (BRIEF) questionnaire and performance-based EF tasks as potential predictors of problematic driving outcomes in adolescents. Forty-six adolescent drivers completed the (1) BRIEF, (2) Trail Making Test, (3) Backwards Digit Span, and (4) self-report on 3 problematic driving outcomes: the number of times of having been pulled over by a police officer, the number of tickets issued, and the number of MVCs. Greater self-reported difficulty with planning and organization was associated with greater odds of having a MVC, whereas inhibition difficulties were associated with greater odds of receiving a ticket. Greater self-reported difficulty across multiple BRIEF subscales was associated with greater odds of being pulled over. Overall findings indicated that the BRIEF, an ecological measure of EF, showed significant association with self-reported problematic driving outcomes in adolescents. No relationship was found between performance-based EF measures and self-reported driving outcomes. The BRIEF could offer unique and quick insight into problematic driving behavior and potentially be an indicator of driving risk in adolescent drivers during clinical evaluations.


Acetylcholine receptor autoantibody secretion by thymocytes: relationship to myasthenia gravis.
We tested thymus cells from 119 patients with acquired myasthenia gravis (MG) for in vitro production of acetylcholine receptor (AChR) binding antibodies (Ab); 109 were seropositive, of which 82 (75%) secreted AChRAb in vitro. As noted in earlier studies, thymus cell secretion of AChRAb paralleled serum Ab levels (rho s = 0.503; p < 0.0001; n = 119). Striational Ab secretion also correlated with the patients' serologic status. AChRAb secretion in vitro predicted the secretory activity of thymus cells implanted in severe combined immune deficiency (SCID) mice. Thymocytes from patients treated with corticosteroids made significantly less AChRAb than thymocytes from untreated patients (p < 0.005). Of particular note: (1) AChRAb was secreted by thymocytes from two of five nonimmunosuppressed patients who had generalized MG but who were seronegative for AChR binding autoantibodies; and (2) AChRAb secretion from both thymic and nonthymic sources was documented in a patient with noninvasive thymoma whose first signs of MG appeared 7 months after thymectomy. We conclude that the thymus in a majority of patients with acquired MG is the principal but not the sole reservoir of immunocytes that specifically react with muscle antigens.


The contribution of de novo coding mutations to autism spectrum disorder.
Whole exome sequencing has proven to be a powerful tool for understanding the genetic architecture of human disease. Here we apply it to more than 2,500 simplex families, each having a child with an autistic spectrum disorder. By comparing affected to unaffected siblings, we show that 13% of de novo missense mutations and 43% of de novo likely gene-disrupting (LGD) mutations contribute to 12% and 9% of diagnoses, respectively. Including copy number variants, coding de novo mutations contribute to about 30% of all simplex and 45% of female diagnoses. Almost all LGD mutations occur opposite wild-type alleles. LGD targets in affected females significantly overlap the targets in males of lower intelligence quotient (IQ), but neither overlaps significantly with targets in males of higher IQ. We estimate that LGD mutation in about 400 genes can contribute to the joint class of affected females and males of lower IQ, with an overlapping and similar number of genes vulnerable to contributory missense mutation. LGD targets in the joint class overlap with published targets for intellectual disability and schizophrenia, and are enriched for chromatin modifiers, FMRP-associated genes and embryonically expressed genes. Most of the significance for the latter comes from affected females.


Forgoing treatment at the end of life in 6 European countries.
Modern medicine provides unprecedented opportunities in diagnostics and treatment. However, in some situations at the end of a patient's life, many physicians refrain from using all possible measures to prolong life. We studied the incidence of different types of treatment withheld or withdrawn in 6 European countries and analyzed the main background characteristics. Between June 2001 and February 2002, samples were obtained from deaths reported to registries in Belgium, Denmark, Italy, the Netherlands, Sweden, and Switzerland. The reporting physician was then sent a questionnaire about the medical decision-making process that preceded the patient's death. The incidence of nontreatment decisions, whether or not combined with other end-of-life decisions, varied widely from 6% of all deaths studied in Italy to 41% in Switzerland. Most frequently forgone in every country were hydration or nutrition and medication, together representing between 62% (Belgium) and 71% (Italy) of all treatments withheld or withdrawn. Forgoing treatment estimated to prolong life for more than 1 month was more common in the Netherlands (10%), Belgium (9%), and Switzerland (8%) than in Denmark (5%), Italy (3%), and Sweden (2%). Relevant determinants of treatment being withheld rather than withdrawn were older age (odds ratio [OR], 1.53; 95% confidence interval [CI], 1.31-1.79), death outside the hospital (death in hospital: OR, 0.80; 95% CI, 0.68-0.93), and greater life-shortening effect (OR, 1.75; 95% CI, 1.27-2.39). In all of the participating countries, life-prolonging treatment is withheld or withdrawn at the end of life. Frequencies vary greatly among countries. Low-technology interventions, such as medication or hydration or nutrition, are most frequently forgone. In older patients and outside the hospital, physicians prefer not to initiate life-prolonging treatment at all rather than stop it later.


Assessment of the perceived stress and burden of family caregivers of the head-and-neck cancer patients at a tertiary care cancer center: A cross-sectional study.
Cancer is a major life-threatening disease and has an impact on both patients and their family members. Caring for cancer patients may lead to several levels of stress which may affect their own health as well as their quality of life. To assess the perceived stress and burden of family caregivers of head and neck cancer patients (HNC) attending cancer care centre at a tertiary care centre, Tamil Nadu. To assess the perceived stress and the burden among caregivers of patients with head and neck cancer using the Perceived Stress Scale (PSS) and Caregiver Strain Index (CSI) respectively. A Cross-sectional study was carried out for a period of three months among the caregivers of head and neck cancer patients at a cancer care centre, Madurai. A total of 200 caregivers were selected by Convenience sampling method. Data was collected using a pretested, self-structured, closed-ended questionnaire by face to face interview method. The study population consisted of Caregivers aged 21-60 years, mostly females (80%), spouses (54%), employed (57%) and uneducated (66%). Most of the caregivers were from lower socioeconomic status (66%) and those who are providing care for 1 to 6 months were more in number. In this study, 82% of caregivers reported high caregiver burden (CSI ≥7) and 67% of caregivers reported high stress (PSS ≥ 26 - 40). Caregivers are experiencing significant burden, particularly with respect to their physical and psychological well-being, economic circumstances, social and personal relationships.


Chitinase production by Bacillus subtilis ATCC 11774 and its effect on biocontrol of Rhizoctonia diseases of potato.
Stem canker and black scurf of potato, caused by Rhizoctonia solani, can be serious diseases causing an economically significant damage. Biocontrol activity of Bacillus subtilis ATCC 11774 against the Rhizoctonia diseases of potato was investigated in this study. Chitinase enzyme was optimally produced by B. subtilis under batch fermentation conditions similar to those of the potato-growing soil. The maximum chitinase was obtained at initial pH 8 and 30 °C. In vitro, the lytic action of the B. subtilis chitinase was detected releasing 355 μg GlcNAc ml⁻¹ from the cell wall extract of R. solani and suggesting the presence of various chitinase enzymes in the bacterial filtrate. In dual culture test, the antagonistic behavior of B. subtilis resulted in the inhibition of the radial growth of R. solani by 48.1% after 4 days. Moreover, the extracted B. subtilis chitinase reduced the growth of R. solani by 42.3% when incorporated with the PDA plates. Under greenhouse conditions, application of a bacterial suspension of B. subtilis at 109 cell mL⁻¹ significantly reduced the disease incidence of stem canker and black scurf to 22.3 and 30%, respectively. In addition, it significantly improved some biochemical parameters, growth and tubers yield. Our findings indicate two points; firstly, B. subtilis possesses a good biocontrol activity against Rhizoctonia diseases of potato, secondly, the harmonization and suitability of the soil conditions to the growth and activity of B. subtilis guaranteed a high controlling capacity against the target pathogen.


Factor VIII alloantibody inhibitors: cost analysis of immune tolerance induction vs. prophylaxis and on-demand with bypass treatment.
Development of inhibitors (alloantibodies to exogenous factor VIII) is the most significant treatment complication in patients with haemophilia A. The only proven way to eradicate inhibitors is through immune tolerance induction (ITI), while bypassing agents are typically employed to treat or prevent bleeds in patients with high titre inhibitors. Costs of these approaches have not been well studied. The aim of this study was to compare lifetime costs of treating patients with severe haemophilia A with inhibitors using on-demand or prophylaxis treatment with bypassing agents and ITI. A decision-analytic model was developed to compare the treatment costs and outcomes. Quantitation of the reduction in bleeding events for patients on prophylaxis and after eradication of inhibitors when on ITI and relapse of inhibitors was derived from published studies. Costs were obtained from standard US costing sources and are reported in 2014 US dollars. Costs and outcomes were discounted 3% per annum. Lifetime costs of treating patients with inhibitors are lower for ITI vs. on-demand or prophylaxis. Patients are also projected to live longer, have greater quality-adjusted life-years, and have fewer bleeding events than patients treated on-demand. Treating patients via ITI to eradicate inhibitors may result in lower lifetime costs and greater life-years and quality-adjusted life-years than treating with bypassing agents.


Epithelial-mesenchymal transition can suppress major attributes of human epithelial tumor-initiating cells.
Malignant progression in cancer requires populations of tumor-initiating cells (TICs) endowed with unlimited self renewal, survival under stress, and establishment of distant metastases. Additionally, the acquisition of invasive properties driven by epithelial-mesenchymal transition (EMT) is critical for the evolution of neoplastic cells into fully metastatic populations. Here, we characterize 2 human cellular models derived from prostate and bladder cancer cell lines to better understand the relationship between TIC and EMT programs in local invasiveness and distant metastasis. The model tumor subpopulations that expressed a strong epithelial gene program were enriched in highly metastatic TICs, while a second subpopulation with stable mesenchymal traits was impoverished in TICs. Constitutive overexpression of the transcription factor Snai1 in the epithelial/TIC-enriched populations engaged a mesenchymal gene program and suppressed their self renewal and metastatic phenotypes. Conversely, knockdown of EMT factors in the mesenchymal-like prostate cancer cell subpopulation caused a gain in epithelial features and properties of TICs. Both tumor cell subpopulations cooperated so that the nonmetastatic mesenchymal-like prostate cancer subpopulation enhanced the in vitro invasiveness of the metastatic epithelial subpopulation and, in vivo, promoted the escape of the latter from primary implantation sites and accelerated their metastatic colonization. Our models provide new insights into how dynamic interactions among epithelial, self-renewal, and mesenchymal gene programs determine the plasticity of epithelial TICs.


Coupling of organic and inorganic aerosol systems and the effect on gas-particle partitioning in the southeastern US.
Several models were used to describe the partitioning of ammonia, water, and organic compounds between the gas and particle phases for conditions in the southeastern US during summer 2013. Existing equilibrium models and frameworks were found to be sufficient, although additional improvements in terms of estimating pure-species vapor pressures are needed. Thermodynamic model predictions were consistent, to first order, with a molar ratio of ammonium to sulfate of approximately 1.6 to 1.8 (ratio of ammonium to 2× sulfate, RN/2S ≈ 0.8 to 0.9) with approximately 70% of total ammonia and ammonium (NH x ) in the particle. Southeastern Aerosol Research and Characterization Network (SEARCH) gas and aerosol and Southern Oxidant and Aerosol Study (SOAS) Monitor for AeRosols and Gases in Ambient air (MARGA) aerosol measurements were consistent with these conditions. CMAQv5.2 regional chemical transport model predictions did not reflect these conditions due to a factor of 3 overestimate of the nonvolatile cations. In addition, gas-phase ammonia was overestimated in the CMAQ model leading to an even lower fraction of total ammonia in the particle. Chemical Speciation Network (CSN) and aerosol mass spectrometer (AMS) measurements indicated less ammonium per sulfate than SEARCH and MARGA measurements and were inconsistent with thermodynamic model predictions. Organic compounds were predicted to be present to some extent in the same phase as inorganic constituents, modifying their activity and resulting in a decrease in [H+]air (H+ in μgm-3 air), increase in ammonia partitioning to the gas phase, and increase in pH compared to complete organic vs. inorganic liquid-liquid phase separation. In addition, accounting for nonideal mixing modified the pH such that a fully interactive inorganic-organic system had a pH roughly 0.7 units higher than predicted using traditional methods (pH = 1.5 vs. 0.7). Particle-phase interactions of organic and inorganic compounds were found to increase partitioning towards the particle phase (vs. gas phase) for highly oxygenated (O : C≥0.6) compounds including several isoprene-derived tracers as well as levoglu-cosan but decrease particle-phase partitioning for low O: C, monoterpene-derived species.


Repertoire of porcine microRNAs in adult ovary and testis by deep sequencing.
MicroRNAs (miRNAs), a large family of short endogenous RNAs known to post-transcriptionally repress gene expression, participate in the regulation of almost every cellular process. Changes in miRNA expression are associated with many pathologies. Ovarian folliculogenesis and testicular spermatogenesis are complex and coordinated biological processes, in which tightly regulated expression and interaction of a multitude of genes could be regulated by these miRNAs. Identification and preliminary characterization of gonad-specific miRNAs would be a prerequisite for a thorough understanding of the role that miRNA-mediated posttranscriptional gene regulation plays in mammalian reproduction. Here, we present the identification of a repertoire of porcine miRNAs in adult ovary and testis using deep sequencing technology. A bioinformatics pipeline was developed to distinguish authentic mature miRNA sequences from other classes of small RNAs represented in the sequencing data. Using this approach, we detected 582 precursor hairpins (pre-miRNAs) encoding for 732 mature miRNAs, of which 673 are unique. Statistically, 224 unique miRNAs (out of 673, 33.28%) were identified which had significant differential expression (DE) between ovary and testis libraries (P < 0.001). Most of DE miRNAs located on the X chromosome (X-linked miRNAs) (24 out of 34, 70.59%) significantly up-regulated in ovary versus testis (P < 0.001). Predictably, X-linked miRNAs are expressed in a testis-preferential or testis-specific pattern. To explore the potential for co-expression among genomic location clusters of X-linked miRNAs, we surveyed the relationship between the distance separating miRNA loci and the coordinate expression patterns of 32 high confidence X-linked miRNAs in seven normal pig tissues using the real-time quantitative PCR (q-PCR) approach. Our results show that proximal pairs of miRNAs are generally co-expressed implying that miRNAs within 50 kb of genomic bases are typically derived from a common transcript. The present study characterizes the miRNA transcriptome of adult porcine gonads, with an emphasis on the co-expression patterns of X-linked miRNAs. Our report should facilitate studies of the organ-specific reproductive roles of miRNAs.


Parents bereaved by offspring suicide: a population-based longitudinal case-control study.
CONTEXT Suicide bereavement remains understudied and poorly understood. OBJECTIVES To examine outcomes of parents bereaved by the suicide death of their offspring and to compare these with both nonbereaved parent controls and parents who had offspring die in a motor vehicle crash (MVC). DESIGN Population-based case-control study. Suicide-bereaved parents were compared with nonbereaved matched control parents in the general population (n = 1415) and with MVC-bereaved parents (n = 1132) on the rates of physician-diagnosed mental and physical disorders, social factors, and treatment use in the 2 years after death of the offspring. Adjusted relative rates (ARRs) were generated by generalized estimating equation models and adjusted for confounding factors. SETTING Manitoba, Canada. PARTICIPANTS All identifiable parents who had an offspring die by suicide between 1996 and 2007 (n = 1415). MAIN OUTCOME MEASURES Mental and physical disorders, social factors, and treatment use. RESULTS Suicide bereavement was associated with an increased rate of depression (ARR, 2.14; 95% CI, 1.88-2.43), anxiety disorders (ARR, 1.41; 95% CI, 1.24-1.60), and marital breakup (ARR, 1.18; 95% CI, 1.13-1.23) in the 2 years after the suicide of an offspring, as compared with the 2 years prior to the death. Suicide-bereaved and MVC-bereaved parents had very few differences on predeath to postdeath outcomes. Depression rate increases were greater for MVC-bereaved parents (19.9%) compared with suicide-bereaved parents (15.9%; P = .005), whereas suicide-bereaved parents had higher rate increases of hospitalization for mental illness (P = .049). Suicide-bereaved parents were more likely than their MVC-bereaved counterparts to have depression (ARR, 1.30; 95% CI, 1.06-1.61), physical disorders (ARR, 1.32; 95% CI, 1.19-1.45), and low income (ARR, 1.34; 95% CI, 1.18-1.51) before their offspring's death. CONCLUSIONS Suicide bereavement is associated with adverse mental health and social outcomes. These consequences appear similar to those associated with MVC bereavement. Parents who lose offspring to suicide appear to be a vulnerable group even prior to their offspring's death.


Q fever endocarditis: is it always subacute or chronic?
Q fever is a zoonotic disease caused by Coxiella burnetii, an obligate intracellular bacterium, which cannot be grown using routine blood culture methods. Although C. burnetii is reported to be the causative agent in approximately 50% of blood culture-negative infective endocarditis cases in developed countries, the incidence in Turkey is yet to be defined. The clinical course of Q fever endocarditis is generally subacute and chronic; the disease may be present for years with only subtle symptoms and no vegetation visible on echocardiography while the bacteria gradually destroy the heart valves. This is the case of the successful treatment of a young man with Q fever endocarditis that had an acute clinical course. In 1 month, he developed New York Heart Association class IV heart failure and a large, 3-cm vegetation was observed on an echocardiogram.


Natural history of celiac disease autoimmunity in a USA cohort followed since 1974.
The natural history and the possible changes of celiac disease (CD) prevalence over time are still unclear. 1) To establish whether loss of tolerance to gluten may occur at any age; 2) to investigate possible changes of CD prevalence over time; and 3) to investigate CD-related co-morbidities. We analyzed 3,511 subjects with matched samples from 1974 (CLUE I) and 1989 (CLUE II). To avoid a selection bias regarding survival, we also screened 840 CLUE I participants who deceased after the 1974 survey. Outcome measure. CD autoimmunity (positivity to auto-antibodies) over time. CD autoimmunity was detected in seven subjects in 1974 (prevalence 1:501) and in an additional nine subjects in 1989 (prevalence 1:219). Two cases of CD autoimmunity were found among the 840 subjects deceased after CLUE I. Compared to controls, untreated CD subjects showed increased incidence of osteoporosis and associated autoimmune disorders, but they did not reach statistical significance. During a 15-year period CD prevalence increased 2-fold in the CLUE cohort and 5-fold overall in the US since 1974. The CLUE study demonstrated that this increase was due to an increasing number of subjects that lost the immunological tolerance to gluten in their adulthood.


Regional fat changes induced by localized muscle endurance resistance training.
The purpose of this study was to examine the effects of a localized muscle endurance resistance training program on total body and regional tissue composition. Seven men and 4 women (aged 23 ± 1 years) were trained with their nondominant leg during 12 weeks, 3 sessions per week. Each session consisted of 1 set of 960-1,200 repetitions (leg press exercise), at 10-30% 1 repetition maximum. Before and after training, body mass, bone mass, bone mineral density (BMD), lean mass, fat mass, and fat percentage were determined by dual-emission x-ray absorptiometry. Energy intakes were registered using a food recall questionnaire. At the whole-body level, body mass, bone mass, BMD, lean mass, or body fat percentage were not significantly changed. However, body fat mass significantly decreased by 5.1% (preexercise: 13.5 ± 6.3 kg; postexercise: 12.8 ± 5.4 kg, p < 0.05). No significant changes in bone mass, lean mass, fat mass, or fat percentage were observed in both the control and trained leg. A significant (p < 0.05) decrease in fat mass was observed in the upper extremities and trunk (10.2 and 6.9%, respectively, p < 0.05). The reduction of fat mass in the upper extremities and trunk was significantly greater (p < 0.05) than the fat mass change observed in the trained leg but not in the control leg. No significant changes were observed in energy intake pre- and postexercise intervention (2,646 ± 444 kcal·d-1 and 2,677 ± 617 kcal·d-1, respectively). In conclusion, the training program was effective in reducing fat mass, but this reduction was not achieved in the trained body segment. The present results expand the limited knowledge available about the plastic heterogeneity of regional body tissues when a localized resistance training program is applied.


Cars, Guns, Aging, and "Giving Up the Keys".
Age-associated changes can impair abilities for safe driving and the use of firearms. We sought to examine multiple perspectives on reducing access to firearms, including similarities and differences compared to reducing driving. Online focus groups and 1-on-1 interviews were conducted (November 2020 to May 2021) in the United States with: older adults who drove and owned firearms; family members of older adult firearm owners/drivers; professionals in aging-related agencies; and firearm retailers/instructors. Recorded sessions were transcribed, coded, and analyzed following a mixed inductive-deductive thematic analysis process. Among 104 participants (81 in focus groups, 23 in interviews), 50 (48%) were female, and 92 (88%) White. Key similarities: decisions are emotional and challenging; needs change over time; safety concerns are heightened by new impairments; prior experiences prompt future planning; tension between autonomy and reliance on trusted others; and strategies like reframing may ease transitions and avoid confrontations. Key differences: "retirement" was not an acceptable term for firearms; reducing driving may affect daily independence more, but there are few alternatives for the psychological safety conferred by firearms; and there are specific firearm-related legal concerns but more driving-related regulations, policies, and resources. The similarities and differences in the processes and preferences related to reducing driving or firearm access have implications for the development of resources to support planning and action. Such resources for the public and providers might empower older adults and their families to make voluntary, shared decisions, and reduce injuries and deaths.


Effects of home-based diet and exercise on functional outcomes among older, overweight long-term cancer survivors: RENEW: a randomized controlled trial.
Five-year survival rates for early stage colorectal, breast, and prostate cancer currently exceed 90% and are increasing. Cancer survivors are at greater risk for second malignancies, other comorbidities, and accelerated functional decline. Lifestyle interventions may provide benefit, but it is unknown whether long-term cancer survivors can modify their lifestyle behaviors sufficiently to improve functional status. To determine whether a telephone counseling and mailed print material-based diet and exercise intervention is effective in reorienting functional decline in older, overweight cancer survivors. Randomized controlled trial of 641 overweight (body mass index > or = 25 and < 40), long-term (> or = 5 years) survivors (aged 65-91 years) of colorectal, breast, and prostate cancer, who were randomly assigned to an intervention group (n = 319) or delayed intervention (control) group (n = 322) in Canada, the United Kingdom, and 21 US states. Individuals were recruited for the Reach out to Enhance Wellness (RENEW) trial from July 1, 2005, through May 17, 2007. A 12-month, home-based tailored program of telephone counseling and mailed materials promoting exercise, improved diet quality, and modest weight loss. The control group was wait-listed for 12 months. Change in self-reported physical function on the Short-Form 36 physical function subscale (score range, 0-100; a high score indicates better functioning) from baseline to 12 months was the primary end point. Secondary outcomes included changes in function on the basic and advanced lower extremity function subscales of the Late Life Function and Disability Index (score range, 0-100), physical activity, body mass index, and overall health-related quality of life. The mean baseline Short-Form 36 physical function score was 75.7. At the 12-month follow-up, the mean function scores declined less rapidly in the intervention group (-2.15; 95% confidence interval [CI], -0.36 to -3.93) compared with the control group (-4.84; 95% CI, -3.04 to -6.63) (P = .03). The mean baseline basic lower extremity function score was 78.2. The mean changes in basic lower extremity function were 0.34 (95% CI, -0.84 to 1.52) in the intervention group compared with -1.89 (95% CI, -0.70 to -3.09) in the control group (P = .005). Physical activity, dietary behaviors, and overall quality of life increased significantly in the intervention group compared with the control group, and weight loss also was greater (2.06 kg [95% CI, 1.69 to 2.43 kg] vs 0.92 kg [95% CI, 0.51 to 1.33 kg], respectively; P < .001). Among older, long-term survivors of colorectal, breast, and prostate cancer, a diet and exercise intervention reduced the rate of self-reported functional decline compared with no intervention. clinicaltrials.gov Identifier: NCT00303875.


HU-331 and Oxidized Cannabidiol Act as Inhibitors of Human Topoisomerase IIα and β.
Topoisomerase II is a critical enzyme in replication, transcription, and the regulation of chromatin topology. Several anticancer agents target topoisomerases in order to disrupt cell growth. Cannabidiol is a major non-euphoriant, pharmacologically active component of cannabis. Previously, we examined the cannabidiol derivative HU-331 in order to characterize the mechanism of the compound against topoisomerase IIα. In this current work, we explore whether cannabidiol (CBD) impacts topoisomerase II activity, and we additionally examine the activity of these compounds against topoisomerase IIβ. CBD does not appear to strongly inhibit DNA relaxation and is not a poison of topoisomerase II DNA cleavage. However, oxidation of CBD allows this compound to inhibit DNA relaxation by topoisomerase IIα and β without poisoning DNA cleavage. Additionally, we found that oxidized CBD, similar to HU-331, inhibits ATP hydrolysis and can result in inactivation of topoisomerase IIα and β. We also determined that oxidized CBD and HU-331 are both able to stabilize the N-terminal clamp of topoisomerase II. Taken together, we conclude that while CBD does not have significant activity against topoisomerase II, both oxidized CBD and HU-331 are active against both isoforms of topoisomerase II. We hypothesize that oxidized CBD and HU-331 act against the enzyme through interaction with the N-terminal ATPase domain. According to the model we propose, topoisomerase II inactivation may result from a decrease in the ability of the enzyme to bind to DNA when the compound is bound to the N-terminus.


microRNA target predictions across seven Drosophila species and comparison to mammalian targets.
microRNAs are small noncoding genes that regulate the protein production of genes by binding to partially complementary sites in the mRNAs of targeted genes. Here, using our algorithm PicTar, we exploit cross-species comparisons to predict, on average, 54 targeted genes per microRNA above noise in Drosophila melanogaster. Analysis of the functional annotation of target genes furthermore suggests specific biological functions for many microRNAs. We also predict combinatorial targets for clustered microRNAs and find that some clustered microRNAs are likely to coordinately regulate target genes. Furthermore, we compare microRNA regulation between insects and vertebrates. We find that the widespread extent of gene regulation by microRNAs is comparable between flies and mammals but that certain microRNAs may function in clade-specific modes of gene regulation. One of these microRNAs (miR-210) is predicted to contribute to the regulation of fly oogenesis. We also list specific regulatory relationships that appear to be conserved between flies and mammals. Our findings provide the most extensive microRNA target predictions in Drosophila to date, suggest specific functional roles for most microRNAs, indicate the existence of coordinate gene regulation executed by clustered microRNAs, and shed light on the evolution of microRNA function across large evolutionary distances. All predictions are freely accessible at our searchable Web site http://pictar.bio.nyu.edu.


Individual quality and personality: bolder males are less fecund in the hermit crab Pagurus bernhardus.
One explanation for animal personality is that different behavioural types derive from different life-history strategies. Highly productive individuals, with high growth rates and high fecundity, are assumed to live life at a fast pace showing high levels of boldness and risk taking, compared with less productive individuals. Here, we investigate among-individual differences in mean boldness (the inverse of the latency to recover from a startling stimulus) and in the consistency of boldness, in male hermit crabs in relation to two aspects of life-history investment. We assessed aerobic scope by measuring the concentration of the respiratory pigment haemocyanin, and we assessed fecundity by measuring spermatophore size. First, we found that individuals investing in large spermatophores also had high concentrations of haemocyanin. Using doubly hierarchical-generalized linear models to analyse longitudinal data on startle responses, we show that hermit crabs vary both in their mean response durations and in the consistency of their behaviour. Individual consistency was unrelated to haemocyanin concentration or spermatophore size, but mean startle response duration increased with spermatophore size. Thus, counter to expectations, it was the most risk-averse individuals, rather than the boldest and most risk prone, that were the most productive. We suggest that similar patterns should be present in other species, if the most productive individuals avoid risky behaviour.


New species and new records of Mydidae from the Afrotropical and Oriental regions (Insecta, Diptera, Asiloidea).
New Mydidae species are described from the Afrotropical and Oriental regions including the first records of this family from several countries in eastern Africa (Kenya, Tanzania, and Uganda) and Mauritania in western Africa as well as Nepal and Thailand in Asia. The new species are, Leptomydinae: Leptomydas notossp. n. (south-western India), Leptomydas raptisp. n. (south-central Nepal), Leptomydas tigrissp. n. (north-central Thailand); Syllegomydinae: Mydaselpidini: Mydaselpis ngurumanisp. n. (south-eastern Kenya, north-eastern Tanzania), Vespiodes phaiossp. n. (south-eastern Kenya); Syllegomydinae: Syllegomydini: Syllegomydas (Notobates) astrictussp. n. (Kenya), Syllegomydas (Notobates) heothinossp. n. (Kenya and Uganda), Syllegomydas (Syllegomydas) elachyssp. n. (northern Zimbabwe). Syllegomydas (Syllegomydas) proximus Séguy, 1928 is recorded from western Mauritania and re-described. Syllegomydas (Notobates) dispar (Loew, 1852), which was previously listed as incertae sedis in the Afrotropical Diptera catalogue, is re-described and illustrated based on examination of the type specimens and several additional specimens from Mozambique. Cephalocera annulata Brunetti, 1912 and Syllegomydas bucciferus Séguy, 1928, described from north-eastern India and previously unplaced in the Oriental Diptera catalogue, are newly combined with Leptomydas Gerstaecker, 1868 and together with Leptomydas indianus Brunetti, 1912, also from north-eastern India, placed in Leptomydinae. Comments on the possible synonymy of the genera of Mydaselpidini are made. Illustrations and photographs are provided to support the descriptions and future identification. A provisional dichotomous key to Mydidae genera occurring in eastern Africa (Kenya, Malawi, Mozambique, Somalia, Tanzania, Uganda) and the Oriental Region is provided. Distribution, occurrence in biodiversity hotspots and high-biodiversity wilderness areas, and seasonal incidence are discussed for all species.


A targeted DNA-PKcs-null mutation reveals DNA-PK-independent functions for KU in V(D)J recombination.
The DNA-dependent protein kinase (DNA-PK) consists of Ku70, Ku80, and a large catalytic subunit, DNA-PKcs. Targeted inactivation of the Ku70 or Ku80 genes results in elevated ionizing radiation (IR) sensitivity and inability to perform both V(D)J coding-end and signal (RS)-end joining in cells, with severe growth retardation plus immunodeficiency in mice. In contrast, we now demonstrate that DNA-PKcs-null mice generated by gene-targeted mutation, while also severely immunodeficient, exhibit no growth retardation. Furthermore, DNA-PKcs-null cells are blocked for V(D)J coding-end joining, but retain normal RS-end joining. Finally, while DNA-PK-null fibroblasts exhibited increased IR sensitivity, DNA-PKcs-deficient ES cells did not. We conclude that Ku70 and Ku80 may have functions in V(D)J recombination and DNA repair that are independent of DNA-PKcs.


Contextual and social variables modulate aesthetic appreciation of bodily and abstract art stimuli.
Despite the increasing interest in the plasticity of aesthetic appreciation, we know comparatively little about the role of individuals' cultural (e.g. the appreciators' expertise) and of social emotional-cognitive (e.g. the social influence of people perceived as warm or competent) variables in modulating the appreciation process. In two experiments we investigated 1) whether people with different art-expertise are influenced differently by contextual (i.e. stimuli primed as art) and social (i.e. stimuli rated as beautiful by art-critics) information and 2) whether acknowledging the judgment of a person perceived as warm or as competent has a different influence on individuals' aesthetic appreciation of art works. Warmth and competence are two social dimensions of fundamental importance for categorizing others as in-group or out-group (Fiske, Cuddy, Glick, & Xu, 2002). We found that insinuating that the observed works were pieces of art, highly appreciated by art critics, lead expert participants to judge the stimuli as more beautiful in comparison to when the very same stimuli were not preceded by any manipulation. Moreover, we found that both art-experts and non-experts rated the stimuli as more beautiful when they believed it to be highly appreciated by people perceived as warm vs people perceived as competent. These results provide novel information on the plasticity of aesthetics and pave the way to understanding how tastes and preferences in the domain of aesthetics can be influenced.