Using Metadynamics to Reveal Extractant Conformational Free Energy Landscapes.
Understanding the impact of extractant functionalization on metal-binding energetics in liquid-liquid extraction is essential to guide the development of better separation processes. Traditionally, computational extractant design uses electronic structure calculations on metal-ligand clusters to determine the metal-binding energy of the lowest energy state. Although highly accurate, this approach does not account for all of the relevant physics encountered under experimental conditions. Such methodologies often neglect entropic contributions such as temperature effects and ligand flexibility, in addition to approximating solvent-extractant interactions with implicit solvent models. In this study, we use classical molecular dynamics simulations with an advanced sampling method, metadynamics, to map out extractant molecule conformational free energies in the condensed phase. We generate the complete conformational landscape in solution for a family of bidentate malonamide-based extractants with different functionalizations of the headgroup and the side chains. In particular, we show how such alkyl functionalization reshapes the free energy landscape, affecting the free energy penalty of organizing the extractant into the cis-like metal-binding conformation from the trans-like conformation of the free extractant in solution. Specifically, functionalizing alkyl tails to the center of the headgroup has a greater influence on increasing molecular rigidity and disfavoring the binding conformation than functionalizing side chains. These findings are consistent with trends in metal-binding energetics based on experimentally reported distribution ratios. We also consider a different bidentate extractant molecule, carbamoylmethylphosphine oxide, and show how the choice of solvent can further reshape the conformational energetic landscape. This study demonstrates the feasibility of using molecular dynamics simulations with advanced sampling techniques to investigate extractant conformational energetics in solution, which, more broadly, will enable extractant design that accounts for entropic effects and explicit solvation.


Physician responses to global physician expenditure budgets in Canada: a common property perspective.
Global expenditure budgets in the fee-for-service physician sector create management problems for both funders and physicians. Global expenditure cap policies must be designed, and appropriate institutional structures created, to mitigate perverse utilization incentives, manage collective utilization, and diffuse the internal professional and the funder-profession tensions created by a capped budget. Two Canadian provinces that adopted different approaches to the design of their physician expenditure cap policies experienced different outcomes in utilization growth. The outcomes, however, are the opposite to what one would predict based on an analysis of the incentive structures embodied in the two provinces' policies. An analytic framework developed for the study of common-property resources is applied to the differing physician responses to global budgets across the two provinces. The insights offered by this framework can guide policy design for global physician budgets, and they indicate the critical importance of physician acceptance of such a policy.


Role of Phosphodiesterase 1 in the Regulation of Real-Time cGMP Levels and Contractility in Adult Mouse Cardiomyocytes.
In mouse cardiomyocytes, the expression of two subfamilies of the calcium/calmodulin-regulated cyclic nucleotide phosphodiesterase 1 (PDE1)-PDE1A and PDE1C-has been reported. PDE1C was found to be the major subfamily in the human heart. It is a dual substrate PDE and can hydrolyze both 3',5'-cyclic adenosine monophosphate (cAMP) and 3',5'-cyclic guanosine monophosphate (cGMP). Previously, it has been reported that the PDE1 inhibitor ITI-214 shows positive inotropic effects in heart failure patients which were largely attributed to the cAMP-dependent protein kinase (PKA) signaling. However, the role of PDE1 in the regulation of cardiac cGMP has not been directly addressed. Here, we studied the effect of PDE1 inhibition on cGMP levels in adult mouse ventricular cardiomyocytes using a highly sensitive fluorescent biosensor based on Förster resonance energy transfer (FRET). Live-cell imaging in paced and resting cardiomyocytes showed an increase in cGMP after PDE1 inhibition with ITI-214. Furthermore, PDE1 inhibition and PDE1A knockdown amplified the cGMP-FRET responses to the nitric oxide (NO)-donor sodium nitroprusside (SNP) but not to the C-type natriuretic peptide (CNP), indicating a specific role of PDE1 in the regulation of the NO-sensitive guanylyl cyclase (NO-GC)-regulated cGMP microdomain. ITI-214, in combination with CNP or SNP, showed a positive lusitropic effect, improving the relaxation of isolated myocytes. Immunoblot analysis revealed increased phospholamban (PLN) phosphorylation at Ser-16 in cells treated with a combination of SNP and PDE1 inhibitor but not with SNP alone. Our findings reveal a previously unreported role of PDE1 in the regulation of the NO-GC/cGMP microdomain and mouse ventricular myocyte contractility. Since PDE1 serves as a cGMP degrading PDE in cardiomyocytes and has the highest hydrolytic activities, it can be expected that PDE1 inhibition might be beneficial in combination with cGMP-elevating drugs for the treatment of cardiac diseases.


Ready-to-use thermally stable mastermix pills for molecular biology applications.
Rolling circle amplification (RCA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification (LAMP), are powerful tools that can be used for gene manipulation, pathogen detection, and infectious disease diagnostics. However, these techniques require trained personnel, as the pipetting steps involved can lead to contamination and, consequently, erroneous results. Furthermore, many of the reagents used in molecular biology are thermally labile and must be kept within a cold-chain. In this article, we present a simple and cost-effective method that allows molecular biology reagents to be thermally stabilized into ready-to-use mastermixes via drying in pullulan and trehalose films. Our experimental results demonstrate that this method is capable of preserving the activity of RCA, PCR, LAMP, ligase, polynucleotide kinase, and Klenow fragment mastermixes for at least 3 months at ambient conditions. Thus, stabilizing reagents via drying in pullulan and trehalose film may allow for a drastic reduction in the number of pipetting steps and the elimination of the need for a cold chain. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2764, 2019.


[Autoimmune hemolytic anemia and posthepatitis-C liver cirrhosis].
Here we are presenting the case of a 70-years-old woman who has hepatic cirrhosis anti-HCV and insulin-dependent diabetes mellitus, without relevant epidemiologic ascendants or previous transfusions and HBV, HIV negatives. On admission to our hospital she showed signs of autoimmune hemolytic anaemia (AHA) which was confirmed by positive direct Coombs test and an improvement in blood test after corticoid treatment. Having discarded other possible causes such as drugs infectious diseases or essential mixed cryoglobulinemia (CME), we put forward the possible association between AHA and infection by HCV, where AHA was an extrahepatic immunological manifestation of HCV. This fact has never been brought to light in previous medical literature.


Electroreception in Gymnotus carapo: pre-receptor processing and the distribution of electroreceptor types.
This paper describes the peripheral mechanisms involved in signal processing of self- and conspecific-generated electric fields by the electric fish Gymnotus carapo. The distribution of the different types of tuberous electroreceptor and the occurrence of particular electric field patterns close to the body of the fish were studied. The density of tuberous electroreceptors was found to be maximal on the jaw (foveal region) and very high on the dorsal region of the snout (parafoveal region), decaying caudally. Tuberous type II electroreceptors were much more abundant than type I electroreceptors. Type I electroreceptors occurred exclusively on the head and rostral trunk regions, while type II electroreceptors were found along as much as 90 % of the fish. Electrophysiological data indicated that conspecific- and self-generated electric currents are 'funnelled' by the high conductivity and geometry of the body of the fish. These currents are concentrated at the peri-oral zone, where most electroreceptors are located. Moreover, within this region, field vector directions were collimated, constituting the most efficient stimulus for electroreceptors. It can be concluded that the passive properties of the fish tissue represent a pre-receptor device that enhances exafferent and reafferent electrical signals at the fovea-parafoveal region.


Utility and relationships of biomarkers of smoking in African-American light smokers.
Although expired carbon monoxide (CO) and plasma cotinine (COT) have been validated as biomarkers of self-reported cigarettes per day (CPD) in heavy smoking Caucasians, their utility in light smokers is unknown. Further, variability in CYP2A6, the enzyme that mediates formation of COT from nicotine and its metabolism to trans-3'-hydroxycotinine (3HC), may limit the usefulness of COT. We assessed whether CO and COT are correlated with CPD in African-American light smokers (< or =10 CPD, n = 700), a population with known reduced CYP2A6 activity and slow COT metabolism. We also examined whether gender, age, body mass index, smoking mentholated cigarettes, or rate of CYP2A6 activity, by genotype and phenotype measures (3HC/COT), influence these relationships. At baseline, many participants (42%) exhaled CO of < or =10 ppm, the traditional cutoff for smoking, whereas few (3.1%) had COT below the cutoff of < or =14 ng/mL; thus, COT seems to be a better biomarker of smoking status in this population. CPD was weakly correlated with CO and COT (r = 0.32-0.39, P < 0.001), and those reporting fewer CPD had higher CO/cigarette and COT/cigarette, although the correlations coefficients between these variables were also weak (r = -0.33 and -0.08, P < 0.05). The correlation between CPD and CO was not greatly increased when analyzed by CYP2A6 activity, smoking mentholated cigarettes, or age, although it appeared stronger in females (r = 0.38 versus 0.21, P < 0.05) and obese individuals (r = 0.38 versus 0.24, P < 0.05). Together, these results suggest that CO and COT are weakly associated with self-reported cigarette consumption in African-American light smokers, and that these relationships are not substantially improved when variables previously reported to influence these biomarkers are considered.


The effects of magnetic field distortion on the accuracy of passive device localization frames in MR imaging.
The interventional magnetic resonance (MR) imaging environment presents many challenges for the accurate localization of interventional devices. In particular, geometric distortion of the static magnetic field may be both appreciable and unpredictable. This paper aims to quantify the sensitivity of localization error of various passive device localization frames to static magnetic field distortion in MR. Three localization frames were considered based on having distinctly different methods of encoding position and orientation in MR images. For each frame, the effects of static field distortion were modeled, allowing rotational and translational errors to be computed as functions of the level of distortion, which was modeled using a first order approximation. Validation of the model was performed by imaging the localization frames in a 3T clinical MR scanner, and simulating the effects of static field distortion by varying the scanner's center frequency and gradient shim values. Plots of the rotational and translational components of error in localization frame position and orientation estimates are provided for ranges of uniform static field distortions of 1-100 μT and static field distortion gradients of 0.01-1 mT/m in all three directions. The theoretical estimates are in good agreement with the results obtained by imaging. The error in position and orientation estimation of passive localization frames in MR can be sensitive to static magnetic field distortions. The level of sensitivity, the type of error (i.e., rotational or translational), and the direction of error are dependent on the frame's design and the method used to image it. If 2D gradient echo imaging is employed, frames with position and orientation estimate sensitivity to slice-select error (such as the z-frame) should be avoided, since this source of error is not easily correctable. Accurate frame position and orientation estimates that are insensitive to static field distortion can be achieved using 2D gradient echo imaging if: (a) the method of determining position and orientation only uses in-plane measurements of marker positions, (b) the in-plane marker positions in images are not sensitive to slice-select error, and (c) methods of correcting in-plane error in the frequency-encoded direction are employed.


Regulation of Arabidopsis tapetum development and function by DYSFUNCTIONAL TAPETUM1 (DYT1) encoding a putative bHLH transcription factor.
In flowering plants, male fertility depends on proper cell differentiation in the anther. However, relatively little is known about the genes that regulate anther cell differentiation and function. Here, we report the analysis of a new Arabidopsis male sterile mutant, dysfunctional tapetum1 (dyt1). The dyt1 mutant exhibits abnormal anther morphology beginning at anther stage 4, with tapetal cells that have excess and/or enlarged vacuoles and lack the densely stained cytoplasm typical of normal tapetal cells. The mutant meiocytes are able to complete meiosis I, but they do not have a thick callose wall; they often fail to complete meiotic cytokinesis and eventually collapse. DYT1 encodes a putative bHLH transcription factor and is strongly expressed in the tapetum from late anther stage 5 to early stage 6, and at a lower level in meiocytes. In addition, the level of DYT1 mRNA is reduced in the sporocyteless/nozzle (spl/nzz) and excess microsporocytes1/extra sporogenous cell (ems1/exs) mutants; together with the mutant phenotypes, this suggests that DYT1 acts downstream of SPL/NZZ and EMS1/EXS. RT-PCR results showed that the expression levels of many tapetum-preferential genes are reduced significantly in the dyt1 mutant, indicating that DYT1 is important for the expression of tapetum genes. Our results support the hypothesis that DYT1 is a crucial component of a genetic network that controls anther development and function.


[Improving clinical examination of workers in metallurgic enterprises].
The authors justify necessity to improve system of clinical examination of workers, within "Health" priority national project, present suggestions on realization of specific measures in metallurgic enterprises.


A model-based analysis: what potential could there be for a S. aureus vaccine in a hospital setting on top of other preventative measures?
Over the past decade, there has been sustained interest and efforts to develop a S. aureus vaccine. There is a need to better evaluate the potential public health impact of S. aureus vaccination, particularly given that preventative measures exist to reduce infection. To our knowledge, there is no previous work to assess the potential of a S. aureus vaccine to yield additional MRSA infection reduction in a hospital setting, on top of other preventative measures that already proved efficient. The main objectives were to propose a versatile simulation framework for assessing potential added benefits of a hypothetical S. Aureus vaccine in conjunction with other preventative measures, and to illustrate possibilities in a given hospital setting. To this end, we employed a recently published dynamic transmission modelling framework that we further adapted and expanded to include a hypothetical S. aureus vaccination component in order to estimate potential benefits of vaccinating patients prior to hospital admission. Model-based projections indicate that even with other hygiene prevention measures in place, vaccination of patients prior to hospital admission has the potential to provide additional reduction of MRSA infection. Vaccine coverage and vaccine efficacy are key factors that would ultimately impact the magnitude of this reduction. For example, in an average case scenario with 50% decolonization, 50% screening and 50% hygiene compliance level in place, S. aureus vaccination with 25% vaccine coverage, 75% vaccine efficacy against infection, and 0% vaccine efficacy against colonization, may lead to 12% model-projected additional reduction in MRSA infection prevalence due to vaccination, while this reduction could reach 37% for vaccination with 75% vaccine coverage and 75% vaccine efficacy against infection in the same average case scenario. S. aureus vaccination could potentially provide additional reduction of MRSA infection in a hospital setting, on top of reductions from hygiene prevention measures. The magnitude of such additional reductions can vary significantly depending on the level of hygiene prevention measures in place, as well as key vaccine factors such as coverage and efficacy. Identifying appropriate combinations of preventative measures may lead to optimal strategies to effectively reduce MRSA infection in hospitals.


Relative Effects of Minoxidil 5%, Platelet-Rich Plasma, and Microneedling in Pattern Hair Loss: A Systematic Review and Network Meta-Analysis.
Combination treatments may improve the utility of approved agents for the treatment of pattern hair loss (PHL); however, head-to-head comparisons are lacking. The aim of the study was to compare the efficacy of 5% minoxidil, platelet-rich plasma (PRP), and microneedling across adults with PHL insofar as change in total hair density at 24 weeks. We conducted a literature search in July 2022. Through our Bayesian network meta-analysis, we estimated treatments' surface under the cumulative ranking distribution (SUCRA) values and relative effects - in terms of mean difference (MD). Data from 27 trials, totaling 1,110 patients, were extracted. Interventions were ranked based on the probability of inducing hair density improvements: 5% minoxidil plus microneedling (SUCRA = 95.8%), 5% minoxidil plus PRP (SUCRA = 64.7%), 5% minoxidil (SUCRA = 53.9%), PRP (SUCRA = 34.9%), microneedling (SUCRA = 27.8%), and PRP with microneedling (SUCRA = 22.9%). The efficacy of 5% minoxidil plus microneedling in improving total hair density was significantly greater (p < 0.05) than 5% minoxidil monotherapy (MD = 13 hairs/cm2), PRP monotherapy (MD = 16 hairs/cm2), and microneedling monotherapy (MD = 17 hairs/cm2). Five percent minoxidil plus microneedling is an effective treatment option for improving hair density at 6 months in adult PHL patients.


Egg corticosterone: a noninvasive measure of stress in egg-laying birds.
Quantitative measures of corticosteroids in biological samples that can be obtained noninvasively, such as saliva, feces, and body hair, have important potential as contributing elements in assessing the quality of captive environments and the severity of experimental procedures. Egg-laying chickens may be of particular interest because the corticosterone contents of the egg may have potential as a convenient measure of preceding adrenocortical activity. To develop methods to reliably quantify corticosterone content in the egg white and yolk, corticosterone content in eggs from 15 egg-laying chickens housed in single production cages were compared with that of eggs from 15 sister chickens, group housed in 1450 cm2 cages equipped with bedding, straw nests, sand baths, and perches. Approximately 80% of the total amount of corticosterone in the eggs was found in the yolk, and there was a positive correlation between yolk corticosterone concentration and total egg corticosterone (r = 0.90, n = 30, P < .001). The egg white contained approximately 20% of the total amount of corticosterone, but there was no correlation between concentrations in the white and the total corticosterone content of the eggs (r = 0.003). There was no difference in the white and yolk corticosterone concentrations or total egg corticosterone between singly housed and group-housed egg-laying hens. Quantitative analyses of corticosterone concentration in eggs may assist when analyzing the stressfulness of experimental procedures and major changes to the birds' environment that affect the activity of the hypothalamus-pituitary-adrenal axis.


Evaluation of a cognitive affective model of physical activity behavior.
Background: To empirically evaluate a cognitive affective model of physical activity. This bidirectional, cyclical model hypotheses that executive control processes directly influence habitual engagement in exercise and also directly subserve the exercise-induced affective response to acute exercise associated with future physical activity. Methods: The present study employed a one-week prospective, multi-site design. Participant recruitment and data collection occurred at two separate University sites (one in the United States and the other in Canada). Participants completed a bout of treadmill exercise, with affect and arousal assessed before, during and after the bout of exercise. Subjective and objective measures of executive function were assessed during this visit. Following this laboratory visit, seven days of accelerometry were employed to measure habitual engagement in physical activity. Results: Within our inactive, young adult sample, we observed some evidence of 1) aspects of executive function were associated with more light-intensity physical activity in the future (1-week later) (r = 0.36, 95% CI = -0.03 to 0.66, P = 0.07), 2) aspects of executive function were associated with post-exercise affect (r = -0.39, 95% CI = -0.67 to -0.03, P = 0.03) and forecasted affect (r =0.47, 95% CI = 0.11 to 0.72, P = 0.01), and 3) aspects of acute exercise arousal and affect were associated with current mild-intensity physical activity behavior (r = 0.41, 95% CI = 0.04 to 0.68, P = 0.03). Conclusion : We demonstrate partial support of a cognitive-affective model of physical activity.


Insights into herpesvirus tegument organization from structural analyses of the 970 central residues of HSV-1 UL36 protein.
The tegument of all herpesviruses contains a capsid-bound large protein that is essential for multiple viral processes, including capsid transport, decapsidation at the nuclear pore complex, particle assembly, and secondary envelopment, through mechanisms that are still incompletely understood. We report here a structural characterization of the central 970 residues of this protein for herpes simplex virus type 1 (HSV-1 UL36, 3164 residues). This large fragment is essentially a 34-nm-long monomeric fiber. The crystal structure of its C terminus shows an elongated domain-swapped dimer. Modeling and molecular dynamics simulations give a likely molecular organization for the monomeric form and extend our findings to alphaherpesvirinae. Hence, we propose that an essential feature of UL36 is the existence in its central region of a stalk capable of connecting capsid and membrane across the tegument and that the ability to switch between monomeric and dimeric forms may help UL36 fulfill its multiple functions.


Effects of mistletoe (Phoradendron villosum) on California oaks.
Mistletoes are a widespread group of plants often considered to be hemiparasitic, having detrimental effects on growth and survival of their hosts. We studied the effects of the Pacific mistletoe, Phoradendron villosum, a member of a largely autotrophic genus, on three species of deciduous California oaks. We found no effects of mistletoe presence on radial growth or survivorship and detected a significant positive relationship between mistletoe and acorn production. This latter result is potentially explained by the tendency of P. villosum to be present on larger trees growing in nitrogen-rich soils or, alternatively, by a preference for healthy, acorn-producing trees by birds that potentially disperse mistletoe. Our results indicate that the negative consequences of Phoradendron presence on their hosts are negligible-this species resembles an epiphyte more than a parasite-and outweighed by the important ecosystem services mistletoe provides.


Genetic risk factors predict disease progression in Crohn's disease patients of the Swiss inflammatory bowel disease cohort.
Crohn's disease (CD) may progress from an inflammatory to a stricturing or penetrating disease phenotype. The aim of our study was to identify single nucleotide polymorphisms (SNPs) that predict disease progression in patients of the Swiss IBD Cohort Study (SIBDCS). We applied a multi-state Markov model for progression behavior of CD with three behavioral states according to the Montreal classification. The model considered transition from B1 to B2/B3 or from B2 to B3 stage. Model dynamics were summarized with transition intensities by including the effect of SNPs and calculating transition intensities for each SNP. We included 1276 CD patients [669 (52.4%) B1, 248 (19.4%) B2, 359 (28.1%) B3 patients] with a median follow-up of 6.8 (interquartile range = 3.6-9.1; range 0-11.6) years. Probability for a B1 patient to develop a stenosis (B1 to B2, q = 0.033) was twice as much as compared to developing a penetrating complication (B3) during the disease course. In contrast, the probability of entering B3 stage was similar regardless of whether antecedent stricture was present (B2 to B3, q = 0.016) or not (B1 to B3, q = 0.016). We identified SNPs within the gene loci encoding ZMIZ1, LOC105373831 and KSR1 as carrying the highest risk for progression to B3, while the presence of SNPs within gene loci TNFSF15 and CEBPB-PTPN1 protected from progression to B2 or B3. We identified new genetic risk factors that can predict disease course in CD patients. A closer understanding on the functional impact of these genetic variations might improve our treatment options finally to prevent disease progression in CD patients.


Renal parathyroid hormone receptors in the chick: downregulation in secondary hyperparathyroid animal models.
We characterized the binding of 125I-[Nle8, Nle18, Tyr34]parathyroid hormone-(1-34) amide [125I-nlPTH-(1-34)] to renal plasma membranes prepared from chicks to determine the effects of secondary hyperparathyroid states on renal PTH receptors. This radioligand exhibited specific binding to membranes with high affinity (Kd, 2-3 X 10(-9) M). Agonists or competitive antagonists of PTH were effective in competing for binding sites labeled with 125I-nlPTH-(1-34), whereas an inactive fragment of PTH, salmon calcitonin, and bovine growth hormone did not compete with the radioligand for renal PTH receptors. Newly hatched chicks raised on control diet with adequate vitamin D and calcium or diets deficient in either vitamin D or calcium were used to study the regulation of renal PTH receptors in experimental models of secondary hyperparathyroidism. We found that both experimental diets resulted in marked hypocalcemia and progressive loss of renal cyclic AMP responsiveness to PTH in vitro. Associated with this refractoriness to the hormone was a marked reduction in PTH receptors in membranes from both vitamin D-deficient and calcium-deficient chick kidney. No change in the affinity of the PTH receptors was found. Vitamin D3, in a single dose of 250 micrograms, partially restored serum calcium of vitamin D-deficient birds toward normal by 72 h and also partly restored renal cyclic AMP responsiveness to PTH and the PTH receptor number toward control values. We conclude that renal refractoriness to PTH observed in experimentally hyperparathyroid animals models is due to a marked loss of plasma membrane receptor sites for PTH without an apparent change in the affinity of the receptors for the hormone.


Improving Medication Adherence Through Adaptive Digital Interventions (iMedA) in Patients With Hypertension: Protocol for an Interrupted Time Series Study.
There is a strong need to improve medication adherence (MA) for individuals with hypertension in order to reduce long-term hospitalization costs. We believe this can be achieved through an artificial intelligence agent that helps the patient in understanding key individual adherence risk factors and designing an appropriate intervention plan. The incidence of hypertension in Sweden is estimated at approximately 27%. Although blood pressure control has increased in Sweden, barely half of the treated patients achieved adequate blood pressure levels. It is a major risk factor for coronary heart disease and stroke as well as heart failure. MA is a key factor for good clinical outcomes in persons with hypertension. The overall aim of this study is to design, develop, test, and evaluate an adaptive digital intervention called iMedA, delivered via a mobile app to improve MA, self-care management, and blood pressure control for persons with hypertension. The study design is an interrupted time series. We will collect data on a daily basis, 14 days before, during 6 months of delivering digital interventions through the mobile app, and 14 days after. The effect will be analyzed using segmented regression analysis. The participants will be recruited in Region Halland, Sweden. The design of the digital interventions follows the just-in-time adaptive intervention framework. The primary (distal) outcome is MA, and the secondary outcome is blood pressure. The design of the digital intervention is developed based on a needs assessment process including a systematic review, focus group interviews, and a pilot study, before conducting the longitudinal interrupted time series study. The focus groups of persons with hypertension have been conducted to perform the needs assessment in a Swedish context. The design and development of digital interventions are in progress, and the interventions are planned to be ready in November 2020. Then, the 2-week pilot study for usability evaluation will start, and the interrupted time series study, which we plan to start in February 2021, will follow it. We hypothesize that iMedA will improve medication adherence and self-care management. This study could illustrate how self-care management tools can be an additional (digital) treatment support to a clinical one without increasing burden on health care staff. ClinicalTrials.gov NCT04413500; https://clinicaltrials.gov/ct2/show/NCT04413500. DERR1-10.2196/24494.


Proliferating cell nuclear antigen expression in papillary thyroid carcinoma.
To correlate the expression of proliferating cell nuclear antigen (PCNA) with various clinicopathological features of papillary thyroid carcinoma. Sections from 131 formalin fixed, paraffin wax embedded papillary thyroid carcinomas were stained with a monoclonal antibody (PC10) directed against PCNA using the avidin-biotin immunoperoxidase (ABC) method. PCNA immunoreactivity was based on the PCNA labelling index (LI) following evaluation of at least 1000 tumour cells, and expressed as follows: grade A (LI < 10%), grade B (10% < or = LI < 25%), and grade C (LI > or = 25%). The relation between PCNA expression in these three groups and other clinicopathological factors, such as sex, age, tumour size, nodal metastases, and histological differentiation, were examined. Based on the labelling index, 57 (43.5%) cases were graded as A, 46 (35.1%) as B, and 28 (21.4%) as C. The female-:male ratios were 6.13:1 for group A, 2.83:1 for group B, and 2.11:1 for group C. The mean (SD) ages of the patients were 39.0 (16.1) in group A, 53.5 (14.4) in group B, and 55.8 (13.3) years in group C. The correlation between age and PCNA grade was strongest in women. PCNA immunoreactivity is correlated with sex and age in patients with papillary thyroid tumours.


Structure of PINK1 and mechanisms of Parkinson's disease-associated mutations.
Mutations in the human kinase PINK1 (hPINK1) are associated with autosomal recessive early-onset Parkinson's disease (PD). hPINK1 activates Parkin E3 ligase activity, involving phosphorylation of ubiquitin and the Parkin ubiquitin-like (Ubl) domain via as yet poorly understood mechanisms. hPINK1 is unusual amongst kinases due to the presence of three loop insertions of unknown function. We report the structure of Tribolium castaneum PINK1 (TcPINK1), revealing several unique extensions to the canonical protein kinase fold. The third insertion, together with autophosphorylation at residue Ser205, contributes to formation of a bowl-shaped binding site for ubiquitin. We also define a novel structural element within the second insertion that is held together by a distal loop that is critical for TcPINK1 activity. The structure of TcPINK1 explains how PD-linked mutations that lie within the kinase domain result in hPINK1 loss-of-function and provides a platform for the exploration of small molecule modulators of hPINK1.


Serum enhancement of human endothelial cell attachment to and spreading on collagens I and IV does not require serum fibronectin or vitronectin.
The initial attachment and spreading of endothelial cells from human umbilical artery onto type I collagen, type IV collagen or gelatin substrata was shown to be enhanced by inclusion of serum in the culture medium. To test whether this serum effect was mediated by adsorption of serum fibronectin or vitronectin onto the collagen, these adhesive glycoproteins were selectively removed from the serum prior to addition to the culture medium. The stimulatory effect of serum on human endothelial cell spreading on collagens I and IV was also observed with serum from which either fibronectin or vitronectin, or both, had been selectively removed. The stimulatory effect for cell spreading on gelatin was diminished by selective removal of serum fibronectin, but unaffected by removal of vitronectin. Human endothelial cell attachment and spreading onto tissue culture plastic was abolished by removal of vitronectin from the serum in the culture medium. These results emphasize that the native structure of collagens is required for serum-enhancement of human endothelial cell attachment and spreading on native collagen types I and IV, and show that on these substrata the stimulated adhesion and spreading are not dependent upon adsorption of serum fibronectin or vitronectin onto the collagen substratum.


Irrigation with potable water versus normal saline in a contaminated musculoskeletal wound model.
Although the use of potable water for wound irrigation is attractive in an austere environment, its effectiveness has not been tested. We sought to compare the effectiveness of potable water irrigation in reducing bacterial number with that of normal saline irrigation. We used an established caprine model involving the creation of a reproducible complex musculoskeletal wound followed by inoculation with luminescent bacteria that allowed for quantitative analysis with a photon-counting camera system. Six hours after injury and inoculation, wound irrigations were performed using pulsatile lavage. Fourteen goats were randomized into two treatment groups: irrigation with 9 L potable water versus irrigation with 9 L normal saline. Images obtained after irrigation were compared with baseline images to determine the reduction in bacterial luminescence resulting from treatment. The irrigation in both groups reduced the bacterial counts by 71% from the preirrigation levels. Potable water reduced the bacterial load as effectively as normal saline in our model.


Trophic actions of recombinant human nerve growth factor on cultured rat embryonic CNS cells.
NGF is a neurotrophic factor for basal forebrain cholinergic neurons and may serve to counteract the cholinergic deficits that are observed in Alzheimer's disease. Prior to the introduction of clinical trials, it is essential that recombinant human NGF (rhNGF) be produced and that its actions on target cells in the CNS be demonstrated. We prepared rhNGF and examined its actions on fetal rat brain neurons in culture including, in particular, the cholinergic neurons of the basal forebrain. rhNGF was more potent in increasing choline acetyltransferase (ChAT) activity in septal cultures than NGF purified from mouse salivary glands (mNGF). ED50s of the beta-NGF dimers were 4.9 pM for rhNGF and 12.4 pM for mNGF. The maximal ChAT activity response was achieved at approximately 35 pM with both NGFs and their efficacies were not significantly different. The two NGFs were not additive in effect. Identical to the results with mNGF, rhNGF strongly enhanced the intensity of ChAT immunostaining in septal cultures. Neither rhNGF nor mNGF affected the appearance of the cultures under phase-contrast illumination. Survival of cells at very low plating density on polyornithine/laminin-coated culture dishes was not affected by rhNGF or mNGF. Protein content and the uptake of GABA were also unaffected. At concentrations of up to 10 micrograms/ml, rhNGF did not significantly increase uptake of dopamine into cultures of ventral mesencephalon. We conclude that rhNGF produces potent and selective actions on cholinergic neurons of the basal forebrain as previously shown for mNGF.


Development and Evaluation of Buccal Films Based on Chitosan for the Potential Treatment of Oral Candidiasis.
In this work, chitosan films were prepared by a casting/solvent evaporation methodology using pectin or hydroxypropylmethyl cellulose to form polymeric matrices. Miconazole nitrate, as a model drug, was loaded into such formulations. These polymeric films were characterized in terms of mechanical properties, adhesiveness, and swelling as well as drug release. Besides, the morphology of raw materials and films was investigated by scanning electron microscopy; interactions between polymers were analyzed by infrared spectroscopy and drug crystallinity studied by differential scanning calorimetry and X-ray diffraction. In addition, antifungal activity against cultures of the five most important fungal opportunistic pathogens belonging to Candida genus was investigated. Chitosan:hydroxypropylmethyl cellulose films were found to be the most appropriate formulations in terms of folding endurance, mechanical properties, and adhesiveness. Also, an improvement in the dissolution rate of miconazole nitrate from the films up to 90% compared to the non-loaded drug was observed. The in vitro antifungal activity showed a significant activity of the model drug when it is loaded into chitosan films. These findings suggest that chitosan-based films are a promising approach to deliver miconazole nitrate for the treatment of candidiasis.


The influence of nanoscale grooved substrates on osteoblast behavior and extracellular matrix deposition.
To fight bone diseases characterized by poor bone quality like osteoporosis and osteoarthritis, as well as in reconstructive surgery, there is a need for a new generation of implantable biomaterials. It is envisioned that implant surfaces can be improved by mimicking the natural extracellular matrix of bone tissue, which is highly a organized nano-composite. In this study we aimed to get a better understanding of osteoblast response to nanometric grooved substrates varying in height, width and spacing. A throughput screening biochip was created using electron beam lithography. Subsequently, uniform large-scale nanogrooved substrates were created using laser interference lithography and reactive ion etching. Results showed that osteoblasts were responsive to nanopatterns down to 75 nm in width and 33nm in depth. SEM and TEM studies showed that an osteoblast-driven calcium phosphate (CaP) mineralization was observed to follow the surface pattern dimensions. Strikingly, aligned mineralization was found on even smaller nanopatterns of 50 nm in width and 17 nm in depth. A single cell based approach for real time PCR demonstrated that osteoblast-specific gene expression was increased on nanopatterns relative to a smooth control. The results indicate that nanogrooves can be a very promising tool to direct the bone response at the interface between an implant and the bone tissue.


Effects of simple imidazoles on human peripheral blood lymphocytes stimulated by mitogen or allogeneic cells.
Five imidazole compounds were added to cultures of human lymphocytes which had been stimulated to undergo blast transformation by exposure to phytohaemagglutinin, pokeweed mitogen or allogeneic cells. Two compounds, clotrimazole and dacarbazine (DTIC) produced a dose related suppression of these responses. Nimorazole was largely inactive whereas metronidazole and tinidazole actually enhanced the response--at least in those cultures stimulated by the plant mitogens. It is suggested that experiments of this kind are helpful in identifying those imidazole compounds that could be used as immunosuppressants in vivo.


Crossing the finish line: follow-up of abnormal test results in a multisite community health center.
Inadequate follow-up of abnormal test results is a common safety problem in outpatient practice. However, it is unclear exactly where and how often failures occur in the results management process. Our goal was to determine where breakdowns occur by examining 4 high-risk abnormal test results in a group of 11 clinics of an urban community health center organization. Using a chart audit, we counted failures in the management of abnormal results of 4 tests: Pap smears, mammograms, international normalized ratio (INR), and prostate-specific antigen (PSA). We assessed documentation that the result was filed in the chart; the provider signed and responded to the result; the patient was notified of the result; the appropriate follow-up occurred, and it occurred in a timely manner or there was explicit patient refusal of the recommended follow-up. There were 344 abnormal test results (105 Pap smears, 82 mammograms, 61 INRs, and 96 PSAs). The highest rate of failures in the management process was at follow-up care; 34% of the abnormal results did not have documentation that appropriate follow-up had occurred (11% for mammography, 26% for INR, 45% for Pap smears, and 46% for PSA). All of the earlier steps were performed with far fewer failures. For patients receiving follow-up care, 49% of the time, follow-up care did not occur in a timely manner. Most breakdowns in the testing process for these 4 abnormal tests were in the final step, documenting that appropriate follow-up care occurred. Office systems for managing abnormal results reporting and patient follow-up are needed to improve the safety and quality of care.


Treatment Patterns Associated with ACR-Recommended Medications in the Management of Fibromyalgia in the United States.
Fibromyalgia (FM) affects up to 6% of U.S. adults, resulting in a significant burden on the health care system and poor quality of life for patients. Duloxetine, pregabalin, and milnacipran are approved for management of FM; however, consensus is lacking regarding optimal therapy. Patients with FM taking approved medications often do not experience meaningful symptom relief, and many experience intolerable adverse events. To assess treatment patterns associated with available and commonly used medications for the management of FM using U.S. health insurance claims. This retrospective analysis used the MarketScan claims database to identify adults with a first diagnosis of FM (ICD-9-CM code 729.1) between 2009 and 2011 with continuous health plan enrollment for 12 months pre- and post-index. Medications of interest were pregabalin, gabapentin, duloxetine, milnacipran, cyclobenzaprine, and tramadol. These are 6 of the 8 medications recommended by the American College of Rheumatology (ACR) for treating FM; the other 2 (amitriptyline and venlafaxine) were only included in some initial assessments. The Charlson Comorbidity Index (CCI) was used to assess overall comorbidity burden. Endpoints included proportion of patients treated within 1 year after first diagnosis; initial treatment pattern; adherence over the first-year follow-up period for the medications of interest; and discontinuation, switching, and combination therapy patterns among pain medications of interest at different time points. Proportion of days covered (PDC; defined as number of days in the period when the patient had drug supply divided by the number of days in the period) was used to define adherence, which was categorized as low (PDC < 50%), medium (PDC 50% to < 80%), or high (PDC ≥ 80%). The time to discontinuation (defined as the first drug supply gap ≥ 90 days) was estimated using Kaplan-Meier analysis. Overall, 240,144 patients met the inclusion criteria. Patients were predominantly women (68%), had preferred provider organization insurance coverage (68%), and had a CCI score < 1 at baseline (69%). Only 31% (n = 74,738) initiated a treatment with a prescription medication listed in the ACR guidelines, and many patients received less than the recommended dose. Most (n = 70,919) patients initially received monotherapy with one of the 8 prescription medications. Of those who started with ≥ 2 medications (n = 3,819), cyclobenzaprine plus tramadol was the most frequent combination. Adherence was suboptimal for all 6 medications of interest. Duloxetine had the highest mean PDC (59%); for all other agents, mean PDC was < 50%. With the exception of duloxetine, discontinuation rates at 6 months were > 50% for all agents. Alterations in therapy were common. Among patients who discontinued their initial treatment of duloxetine, pregabalin, or milnacipran, approximately one-third had switched treatments within 90 days after their first prescription. For those who maintained their initial treatment agent, approximately 50% of patients added a second pain medication within 1 year of treatment initiation. The evidence suggests that patients with FM often do not receive 1 of the prescription medications recommended by ACR guidelines, and those who do are commonly prescribed lower-than-recommended doses, potentially resulting in poor effectiveness and tolerability. Discontinuation, switching, and addition of new pain medications are common, which may indicate low levels of satisfaction with initial treatment. New therapies with improved effectiveness and better tolerability are urgently needed for patients with FM.


Remdesivir Administration in COVID-19 Patients With Renal Impairment: A Systematic Review.
Remdesivir (RDV) is the main antiviral for the treatment of moderate to severe forms of Coronavirus disease 2019 (COVID-19). Several studies revealed a shortening time to clinical improvement of COVID-19 and mortality benefits in patients receiving RDV. The patients with renal disease were excluded from large clinical trials of RDV, and the probable nephrotoxicity of the drug, its metabolites, and the vehicle (sulfobutylether-β-cyclodextrin) have led to the recommendation against using RDV in patients with an estimated glomerular filtration rate of <30 mL/min. This systematic review aimed to collect data about the necessity and safety administration of RDV in the setting of renal impairment. Search through databases including MEDLINE, ScienceDirect, Cochrane Library, and PubMed was performed. The studies were carried out in adults and enrolled patients with different types of renal impairment (ie, acute kidney injury, chronic kidney disease, kidney transplant, and renal replacement therapy) were included. Eligible studies were assessed, and required data were extracted. Twenty-two cross-sectional studies, cohorts, case reports, and case series were included in this review. The mortality rate was between 7.3% and 50%, and various severity of COVID-19 was included in the studies. None of them reported an increase in adverse effects attributed to RDV administration. A decrease in inflammatory mediators and other benefits were obvious. Although the manufacturer's labeling does not recommend RDV administration in patients with severe renal impairment, it seems that nephrotoxicity is less concerning in the population of these patients. Moreover, RDV may be helpful in acute kidney injury induced by the viral invasion of COVID-19. To the best of our knowledge, this is the first systematic review of the use of RDV in kidney failure. Larger, well-designed, and pharmacokinetic studies are required to have a safe and logical recommendation about the use of RDV in patients with renal disorders.


Exploring a scalable real-time simulation for interprofessional education in pharmacy and medicine.
This article was migrated. The article was marked as recommended. The organisational logistics and cost of interprofessional education (IPE) remain barriers to widespread embedding of IPE activities in health care training. The ability of students to undertake IPE activities in the form of treating a virtual patient independently of an instructor and facilities would be of potential benefit. In this study we explore the feasibility and potential IPE benefits of SimPHARM, a cloud based simulation platform that allows students to treat virtual patients in a real-time simulation. We enrolled three pairs of medical and pharmacy students and after familiarisation with the platform assigned them a 2 day virtual patient. At the end of the case they completed a questionnaire, an interview and we reviewed their SimPHARM log-files. The results supported the logistical feasibility of this approach. We found that most students interacted asynchronously on the platform, leaving notes for each other, in order to arrive at their clinical decisions. The participants found their interactions meaningful in helping them to make clinical decisions and learn from each other.


New insights on the potential anti-epileptic effect of metformin: Mechanistic pathway.
Epilepsy is a chronic neurological disease characterized by recurrent seizures. Epilepsy is observed as a well-controlled disease by anti-epileptic agents (AEAs) in about 69%. However, 30%-40% of epileptic patients fail to respond to conventional AEAs leading to an increase in the risk of brain structural injury and mortality. Therefore, adding some FDA-approved drugs that have an anti-seizure activity to the anti-epileptic regimen is logical. The anti-diabetic agent metformin has anti-seizure activity. Nevertheless, the underlying mechanism of the anti-seizure activity of metformin was not entirely clarified. Henceforward, the objective of this review was to exemplify the mechanistic role of metformin in epilepsy. Metformin has anti-seizure activity by triggering adenosine monophosphate-activated protein kinase (AMPK) signalling and inhibiting the mechanistic target of rapamycin (mTOR) pathways which are dysregulated in epilepsy. In addition, metformin improves the expression of brain-derived neurotrophic factor (BDNF) which has a neuroprotective effect. Hence, metformin via induction of BDNF can reduce seizure progression and severity. Consequently, increasing neuronal progranulin by metformin may explain the anti-seizure mechanism of metformin. Also, metformin reduces α-synuclein and increases protein phosphatase 2A (PPA2) with modulation of neuroinflammation. In conclusion, metformin might be an adjuvant with AEAs in the management of refractory epilepsy. Preclinical and clinical studies are warranted in this regard.


Capitation: developing partnerships for health care.
As medical-surgical supply distributors begin to establish partnerships with hospitals, thus sharing financial risk, materiel executives must understand all potential contractual ramifications. This article will discuss the types and features of supply capitated contracts and the subsequent need to correctly assess new categories of contractual risk.


Functional Insights of Salinity Stress-Related Pathways in Metagenome-Resolved Methanothrix Genomes.
Recently, methanogenic archaea belonging to the genus Methanothrix were reported to have a fundamental role in maintaining stable ecosystem functioning in anaerobic bioreactors under different configurations/conditions. In this study, we reconstructed three Methanothrix metagenome-assembled genomes (MAGs) from granular sludge collected from saline upflow anaerobic sludge blanket (UASB) reactors, where Methanothrix harundinacea was previously implicated with the formation of compact and stable granules under elevated salinity levels (up to 20 g/L Na+). Genome annotation and pathway analysis of the Methanothrix MAGs revealed a genetic repertoire supporting their growth under high salinity. Specifically, the most dominant Methanothrix (MAG_279), classified as a subspecies of Methanothrix_A harundinacea_D, had the potential to augment its salinity resistance through the production of different glycoconjugates via the N-glycosylation process, and via the production of compatible solutes as Nε-acetyl-β-lysine and ectoine. The stabilization and reinforcement of the cell membrane via the production of isoprenoids was identified as an additional stress-related pathway in this microorganism. The improved understanding of the salinity stress-related mechanisms of M. harundinacea highlights its ecological niche in extreme conditions, opening new perspectives for high-efficiency methanisation of organic waste at high salinities, as well as the possible persistence of this methanogen in highly-saline natural anaerobic environments. IMPORTANCE Using genome-centric metagenomics, we discovered a new Methanothrix harundinacea subspecies that appears to be a halotolerant acetoclastic methanogen with the flexibility for adaptation in the anaerobic digestion process both at low (5 g/L Na+) and high salinity conditions (20 g/L Na+). Annotation of the recovered M. harundinacea genome revealed salinity stress-related functions, including the modification of EPS glycoconjugates and the production of compatible solutes. This is the first study reporting these genomic features within a Methanothrix sp., a milestone further supporting previous studies that identified M. harundinacea as a key-driver in anaerobic granulation under high salinity stress.


Association between Sleep-Disordered Breathing and Neuropsychological Performance in Older Adults with Mild Cognitive Impairment.
Sleep-disordered breathing in middle-age and older adults has been shown to be linked to a range of neuropsychological deficits, but the extent to which these relationships are evident in older people 'at risk' of developing dementia in unknown. In this study, we aimed to determine whether changes in sleep-disordered breathing and sleep fragmentation during nocturnal sleep were related to neuropsychological dysfunction in patients with mild cognitive impairment. Forty-six patients with MCI (mean age = 66.1 y, sd = 8.4) and 40 age-matched healthy controls (mean age = 63.5 y, sd = 8.9) underwent psychiatric, medical, and neuropsychological assessment, in addition to overnight polysomnography and self-report questionnaires. Measures of hypoxemia, sleep fragmentation, and sleep quality were derived including the apnoea-hypopnea index, oxygen desaturation index, percentage of total sleep time spent below 90% oxygen saturation, arousal index, sleep efficiency, and wake after sleep onset. Patients with MCI did not differ from healthy aging on any measure of sleep-disordered breathing or sleep fragmentation. In MCI, processing speed was negatively correlated with greater sleep time spent below 90% oxygen saturation and a higher apnoea-hypopnea index. In contrast, in the healthy aging, processing speed was negatively correlated with an increased oxygen desaturation index and the arousal index. Sleep-disordered breathing is evident in both healthy aging and MCI with associated decrements in processing speed. Future research is needed to determine the unique and synergistic effects of these differential associations, their potential to inform disease trajectory, and possible therapeutic interventions.


Predictors of impaired heart rate recovery: a myocardial perfusion SPECT study.
Heart rate recovery (HRR) is an established prognostic predictor. However, a number of methodological issues have not been fully explored, including differences in HRR 1 versus 2 min after exercise termination, absolute versus relative HRR, and the impact of beta-blockers. Cross-sectional study. Predictors of impaired absolute and relative HRR 1 (HRR-1, HRR-1%) and 2 min after exercise termination (HRR-2, HRR-2%), defined as their lowest quartiles, were assessed in 1667 patients undergoing cycle exercise myocardial perfusion single photon emission computed tomography, and measures of HRR were compared between patients undergoing myocardial perfusion single photon emission computed tomography with continued, discontinued, and without beta-blockers. Higher resting heart rate was an independent predictor of all measures of impaired HRR (P<0.001 for all). Lower peak heart rate was independently associated with impaired HRR-1, HRR-2, and HRR-2% (P<0.001 for all) but not HRR-1%. Higher summed rest score as a marker of scar and in part left ventricular dysfunction was an independent predictor of impaired HRR-1 (P = 0.010) and HRR-1% (P = 0.025) but not HRR-2 and HRR-2%, whereas lower stroke volume index was an independent predictor of slow HRR-2 (P = 0.004) and HRR-2% (P = 0.02) but not HRR-1 and HRR-1%. HRR-1 (P = 0.98) and HRR-2 (P = 0.86) were similar in patients with continued, discontinued, and without beta-blocker therapy. In contrast, HRR-1% (P = 0.01) and HRR-2% (P = 0.001) were faster in patients on beta-blockers than in the other groups. HRR-1 and HRR-2 as well as HRR-1% and HRR-2% reflect different pathophysiological processes. Relative but not absolute measures of HRR seem to be enhanced under beta-blockers.


[Effect of space flight factors on skeletal muscle ultrastructure].
By electron microscopy ultrastructures of myofibers of the soleus and gastrocnemius muscles of rats flown in zero-g and 1 g were examined. It was found that 4.5--9 hours postflight the soleus of weightless rats showed local destruction of the contractile and mitochondrial systems accompanied by metabolic changes in muscle fibers. Exposure to artificial gravity partially prevented changes in muscle fibers. Examinations of muscle fibers 25 days postflight demonstrated that the changes were reversible. No changes were seen in the mixed -- gastrocnemius -- muscle of both animal groups.


GaAs photovoltaics and optoelectronics using releasable multilayer epitaxial assemblies.
Compound semiconductors like gallium arsenide (GaAs) provide advantages over silicon for many applications, owing to their direct bandgaps and high electron mobilities. Examples range from efficient photovoltaic devices to radio-frequency electronics and most forms of optoelectronics. However, growing large, high quality wafers of these materials, and intimately integrating them on silicon or amorphous substrates (such as glass or plastic) is expensive, which restricts their use. Here we describe materials and fabrication concepts that address many of these challenges, through the use of films of GaAs or AlGaAs grown in thick, multilayer epitaxial assemblies, then separated from each other and distributed on foreign substrates by printing. This method yields large quantities of high quality semiconductor material capable of device integration in large area formats, in a manner that also allows the wafer to be reused for additional growths. We demonstrate some capabilities of this approach with three different applications: GaAs-based metal semiconductor field effect transistors and logic gates on plates of glass, near-infrared imaging devices on wafers of silicon, and photovoltaic modules on sheets of plastic. These results illustrate the implementation of compound semiconductors such as GaAs in applications whose cost structures, formats, area coverages or modes of use are incompatible with conventional growth or integration strategies.


Structure and expression of germline epsilon transcripts in human B cells induced by interleukin 4 to switch to IgE production.
Interleukin 4 (IL-4)-induced IgE production coincides with the appearance of the 2.2-kb productive epsilon-mRNA, but is preceded by synthesis of a 1.7-kb epsilon-RNA. Analysis of cDNA copies of the 5' end of this RNA indicated that the 1.7-kb epsilon-RNA is a germline epsilon immunoglobulin heavy chain transcript with an exon mapping 5' to the switch region. Transcription through switch regions has been implicated in the control of class switching. However, IL-4 or cloned CD4+ T cells were able to induce germline epsilon transcripts without inducing IgE synthesis, for which both signals were required. These results indicate that induction of human germline epsilon-RNA does not necessarily result in IgE synthesis, and that additional regulatory mechanisms are involved in class switching.


Suspension-Induced Stem Cell Transition: A Non-Transgenic Method to Generate Adult Stem Cells from Mouse and Human Somatic Cells.
Adult stem cells (ASCs) can be cultured with difficulty from most tissues, often requiring chemical or transgenic modification to achieve adequate quantities. We show here that mouse primary fibroblasts, grown in suspension, change from the elongated and flattened morphology observed under standard adherent culture conditions of generating rounded cells with large nuclei and scant cytoplasm and expressing the mesenchymal stem cell (MSC) marker (Sca1; Ly6A) within 24 h. Based on this initial observation, we describe here a suspension culture method that, irrespective of the lineage used, mouse fibroblast or primary human somatic cells (fibroblasts, hepatocytes and keratinocytes), is capable of generating a high yield of cells in spheroid form which display the expression of ASC surface markers, circumventing the anoikis which often occurs at this stage. Moreover, mouse fibroblast-derived spheroids can be differentiated into adipogenic and osteogenic lineages. An analysis of single-cell RNA sequence data in mouse fibroblasts identified eight distinct cell clusters with one in particular comprising approximately 10% of the cells showing high levels of proliferative capacity expressing high levels of genes related to MSCs and self-renewal as well as the extracellular matrix (ECM). We believe the rapid, high-yield generation of proliferative, multi-potent ASC-like cells via the process we term suspension-induced stem cell transition (SIST) could have significant implications for regenerative medicine.


The MX-Helix of Muscle nAChR Subunits Regulates Receptor Assembly and Surface Trafficking.
Nicotinic acetylcholine receptors (AChRs) are pentameric channels that mediate fast transmission at the neuromuscular junction (NMJ) and defects in receptor expression underlie neuromuscular disorders such as myasthenia gravis and congenital myasthenic syndrome (CMS). Nicotinic receptor expression at the NMJ is tightly regulated and we previously identified novel Golgi-retention signals in the β and δ subunit cytoplasmic loops that regulate trafficking of the receptor to the cell surface. Here, we show that the Golgi retention motifs are localized in the MX-helix, a juxta-membrane alpha-helix present in the proximal cytoplasmic loop of receptor subunits, which was defined in recent crystal structures of cys-loop receptor family members. First, mutational analysis of CD4-MX-helix chimeric proteins showed that the Golgi retention signal was dependent on both the amphipathic nature of the MX-helix and on specific lysine residues (βK353 and δK351). Moreover, retention was associated with ubiquitination of the lysines, and βK353R and δK351R mutations reduced ubiquitination and increased surface expression of CD4-β and δ MX-helix chimeric proteins. Second, mutation of these lysines in intact β and δ subunits perturbed Golgi-based glycosylation and surface trafficking of the AChR. Notably, combined βK353R and δK351R mutations increased the amount of surface AChR with immature forms of glycosylation, consistent with decreased Golgi retention and processing. Third, we found that previously identified CMS mutations in the ε subunit MX-helix decreased receptor assembly and surface levels, as did an analogous mutation introduced into the β subunit MX-helix. Together, these findings indicate that the subunit MX-helix contributes to receptor assembly and is required for normal expression of the AChR and function of the NMJ. In addition, specific determinants in the β and δ subunit MX-helix contribute to quality control of AChR expression by intracellular retention and ubiquitination of unassembled subunits, and by facilitating the appropriate glycosylation of assembled surface AChR.


Percutaneous Hemodynamic Support (Impella) in Patients with Advanced Heart Failure and/or Cardiogenic Shock Not Eligible to PROTECT II Trial.
PROTECT I and II trials have tested the efficacy of Impella in patents with high-risk percutaneous coronary intervention (PCI). However, patients with severe hemodynamic instability such as cardiac arrest, ST-segment elevated myocardial infarction (STEMI), or cardiogenic shock were excluded. The objective was to investigate the efficacy of Impella in sicker patient population who were not included in PROTECT trials. These patients merit high-risk PCI who had cardiogenic shock and unstable or decompensated heart failure (HF). From December 2010 to March 2012, 10 consecutive patients with extremely high surgical risk and hemodynamic instability underwent urgent PCI with Impella 2.5 support (Abiomed Inc., Danvers, MA). These patients were presented with advance HF and/or cardiogenic shock. Among the 10 included patients, 3 patients were with cardiac arrest and 1 patient was with acute myocardial infarction. All patients had successful Impella implantation and remained hemodynamically stable during high-risk PCI. Among the 10 patients 2 patients (20%) died within 1 month and 1 patient developed limb ischemia. In high-risk population nonrandomizable to PROTECT trials with advance HF/cardiogenic shock, Impella could be an important tool for hemodynamic support to PCI or could be a bridge to left ventricle assist device to achieve good recovery. Larger studies need to be conducted on this high-risk population.


FLIP-ping out: death receptor signaling in the prostate.
Prostate cancer is a leading cause of cancer related death. The growth of normal prostate epithelial cells is under the tight control of various growth factors, most notably androgens, such that castration leads to apoptosis of this cell population. Androgen-depletion has a similar effect on prostate cancers; however, following initial regression tumors often return in an androgen-depletion independent form that is frequently lethal. Thus, castration induced prostate regression in rodents has been a valuable model for identifying cell signaling pathways that control the proliferation and apoptosis of both normal and neoplastic prostate epithelial cells. For example, studies of normal prostate regression demonstrated the critical role of paracrine (stromally produced) transforming growth factor-beta. This review examines the role of the TNF-family death receptors and caspases-8 and -10 in prostate epithelial cell death. There is significant evidence that expression of the caspase-8 inhibitor FLIP (FLICE-like inhibitory protein) is androgen regulated and that this protein is one of the key regulators of androgen withdrawal induced cell death. However, it is not yet known which of the death receptor pathways is required for prostate apoptosis in vivo, and this remains an active topic of research.


Gating Pore Currents in Sodium Channels.
Voltage-gated sodium channels belong to the superfamily of voltage-gated cation channels. Their structure is based on domains comprising a voltage sensor domain (S1-S4 segments) and a pore domain (S5-S6 segments). Mutations in positively charged residues of the S4 segments may allow protons or cations to pass directly through the gating pore constriction of the voltage sensor domain; these anomalous currents are referred to as gating pore or omega (ω) currents. In the skeletal muscle disorder hypokalemic periodic paralysis, and in arrhythmic dilated cardiomyopathy, inherited mutations of S4 arginine residues promote omega currents that have been shown to be a contributing factor in the pathogenesis of these sodium channel disorders. Characterization of gating pore currents in these channelopathies and with artificial mutations has been possible by measuring the voltage-dependence and selectivity of these leak currents. The basis of gating pore currents and the structural basis of S4 movement through the gating pore has also been studied extensively with molecular dynamics. These simulations have provided valuable insight into the nature of S4 translocation and the physical basis for the effects of mutations that promote permeation of protons or cations through the gating pore.


Adhesion-dependent cell mechanosensitivity.
The conversion of physical signals, such as contractile forces or external mechanical perturbations, into chemical signaling events is a fundamental cellular process that occurs at cell-extracellular matrix contacts, known as focal adhesions. At these sites, transmembrane integrin receptors are associated via their cytoplasmic domains with the actin cytoskeleton. This interaction with actin is mediated by a submembrane plaque, consisting of numerous cytoskeletal and signaling molecules. Application of intrinsic or external forces to these structures dramatically affects their assembly and triggers adhesion-mediated signaling. In this review, we discuss the structure-function relationships of focal adhesions and the possible mode of action of the putative mechanosensor associated with them. We also discuss the general phenomenon of mechanosensitivity, and the approaches used to measure local forces at adhesion sites, the cytoskeleton-mediated regulation of local contractility, and the nature of the signaling networks that both affect contractility and are affected by it.


Advances in the field of π-conjugated 2,2':6',2"-terpyridines.
This critical review summarizes the research progress made in the field of π-conjugated terpyridines within the last decade. Supramolecular materials based on metal ion complexes of 2,2':6',2"-terpyridine derivatives have found manifold potential applications-from opto-electronic devices to life science. In this contribution, synthetic strategies towards π-conjugated terpyridines and their incorporation into advanced supramolecular architectures are evaluated. Applications as photoactive species in, e.g., photovoltaic devices, polymer light-emitting diodes (PLEDSs) and nanotechnology are discussed comprehensively (523 references).


Drug induced liver injury is associated with high mortality-A study from a tertiary care hospital in Pakistan.
In light of few established drug induced liver injury (DILI) registries, this study aims to evaluate the clinical spectrum and predictors of mortality and morbidity of hospitalized patients with suspected DILI. DILI cases were identified and categorized on basis of COIMS/RUCAM score and the exclusion of other liver diseases. Clinical and laboratory parameters were analyzed to identify the predictors of morbidity (prolonged hospital stay > 5 days) and mortality. Out of 462 patients, there were 264 (57.6%) males and the mean age of the cohort was 50.83 years (range: 20-94 years). DILI was classified as definite or highly probable in 31.1%, probable in 62.5%, and possible in 7.4% of cases. Pattern of liver injury was hepatocellular in 25.1%, cholestatic in 56.17%, and mixed in 18.72% of patients. Anti-tuberculosis drugs (ATDs) were found to be the most common category of drugs causing DILI, in 295 (63.9%) patients. Clinically, encephalopathy was present in 21.6% patients; other presenting symptoms included abdominal pain (57.1%), vomiting (57.1%), jaundice (54.1%) and pruritus (42.3%). In-hospital mortality was 26.5% and prolonged hospital stay (> 5 days) was observed in 35.93% of patients. Mortality was significantly greater in patients with encephalopathy, male gender, hepatocellular pattern of DILI, increased INR and use of ventilator support. In our study, the most frequent cause of DILI in hospitalized patients was ATDs. More than a quarter of patients died during hospital stay. A close control of clinical and biochemical parameters are required to prevent and monitor DILI, especially in patients taking ATDs in our region.


Facet joints infiltration: a viable alternative treatment to physiotherapy in patients with low back pain due to facet joint arthropathy.
There is a paucity of information on treatment options and outcome of treatment of patients that have MRI findings of FJA presenting with low back pain in the developing country. This prospective cohort study conducted mainly in the University of Abuja Teaching Hospital Gwagwalada, Abuja Nigeria is to compare the short-term clinical outcome between two groups, one having undergone facet joints infiltration (FJI) and the other physiotherapy for facet joint arthropathy (FJA). All patients with clinical lower back pain (LBP) and MRI features of FJA were recruited into this prospective study randomly. All the FJI patients had depot methylprednisolone and the physiotherapy patients had McKenzie regimen. Outcome questionnaires comprising Oswentry disability index scores (ODI), visual analogue scores (VAS), and patient satisfaction scores (PSS) were completed at all clinic visits. Follow-up was for 6 months for both groups. The results were analyzed using SPSS 17.0. There were 10 FJI patients and 8 physiotherapy patients (1.25: 1). The outcome scores comprised the following ODI, VAS, and PSS. The FJI group had a better score compared to the physiotherapy group at short-term evaluation and this difference was statistically significant. The female patients in both groups fare better compared to the male counterparts. FJI offered added benefit over physiotherapy in LBPs diagnosed with FJA. Patients with FJI have a significant reduction in pain symptom than the patients in the physiotherapy group. Though majority of the patients reported satisfactory outcome, the FJI patients group had a better outcome.


Polypathology and dementia after brain trauma: Does brain injury trigger distinct neurodegenerative diseases, or should they be classified together as traumatic encephalopathy?
Neuropathological studies of human traumatic brain injury (TBI) cases have described amyloid plaques acutely after a single severe TBI, and tau pathology after repeat mild TBI (mTBI). This has helped drive the hypothesis that a single moderate to severe TBI increases the risk of developing late-onset Alzheimer's disease (AD), while repeat mTBI increases the risk of developing chronic traumatic encephalopathy (CTE). In this review we critically assess this position-examining epidemiological and case control human studies, neuropathological evidence, and preclinical data. Epidemiological studies emphasize that TBI is associated with the increased risk of developing multiple types of dementia, not just AD-type dementia, and that TBI can also trigger other neurodegenerative conditions such as Parkinson's disease. Further, human post-mortem studies on both single TBI and repeat mTBI can show combinations of amyloid, tau, TDP-43, and Lewy body pathology indicating that the neuropathology of TBI is best described as a 'polypathology'. Preclinical studies confirm that multiple proteins associated with the development of neurodegenerative disease accumulate in the brain after TBI. The chronic sequelae of both single TBI and repeat mTBI share common neuropathological features and clinical symptoms of classically defined neurodegenerative disorders. However, while the spectrum of chronic cognitive and neurobehavioral disorders that occur following repeat mTBI is viewed as the symptoms of CTE, the spectrum of chronic cognitive and neurobehavioral symptoms that occur after a single TBI is considered to represent distinct neurodegenerative diseases such as AD. These data support the suggestion that the multiple manifestations of TBI-induced neurodegenerative disorders be classified together as traumatic encephalopathy or trauma-induced neurodegeneration, regardless of the nature or frequency of the precipitating TBI.


Analysis of the N-terminal region of human MLKL, as well as two distinct MLKL isoforms, reveals new insights into necroptotic cell death.
The pseudokinase mixed lineage kinase domain-like (MLKL) is an essential effector of necroptotic cell death. Two distinct human MLKL isoforms have previously been reported, but their capacities to trigger cell death have not been compared directly. Herein, we examine these two MLKL isoforms, and further probe the features of the human MLKL N-terminal domain that are required for cell death. Expression in HEK293T cells of the N-terminal 201 amino acids (aa) of human MLKL is sufficient to cause cell death, whereas expression of the first 154 aa is not. Given that aa 1-125 are able to initiate necroptosis, our findings indicate that the helix that follows this region restrains necroptotic activity, which is again restored in longer constructs. Furthermore, MLKL isoform 2 (MLKL2), which lacks much of the regulatory pseudokinase domain, is a much more potent inducer of cell death than MLKL isoform 1 (MLKL1) in ectopic expression studies in HEK293T cells. Modelling predicts that a C-terminal helix constrains the activity of MLKL1, but not MLKL2. Although both isoforms are expressed by human monocyte-derived macrophages at the mRNA level, MLKL2 is expressed at much lower levels. We propose that it may have a regulatory role in controlling macrophage survival, either in the steady state or in response to specific stimuli.


A comparison between different survival and threshold models with an application to piglet preweaning survival in a dry-cured ham-producing crossbred line.
Different approaches for predicting genetic merit of piglet preweaning survival were compared using proportional hazard, threshold (TM), and sequential threshold (STM) models. Data were from 13,924 crossbred piglets (1,347 litters), born from 2000 to 2006, and originated by mating 189 Large White C21 Gorzagri boars to 328 Large White-derived crossbred sows. A frailty proportional hazard model was fitted assuming 2 different baseline hazard functions (Cox and Weibull time-dependent model) and including sire and nursed litter as random effects. The TM and STM included the same effects as considered in the proportional hazard model. Model fitting was evaluated in terms of goodness of fit and predictive ability. The goodness-of-fit was evaluated using the local weighted regression and the mean squared error, whereas the predictive ability was assessed by using a cross-validation procedure. Estimated sire variances for piglet preweaning mortality were low, and heritability ranged from 0.04 to 0.06. All 4 models led to similar ranking of sires. Results suggest that STM may be preferred to the other models for genetic evaluation of piglet preweaning survival, both for its better predictive ability and its easier interpretation. Further, STM is computationally less demanding than survival models and allows for estimating different variance components from birth up to weaning.


Computer generated hologram with characteristics of reflection: reflectance distributions and reflected images.
This paper proposes a new computer generated hologram (CGH) method that considers the reflectance distribution on object surfaces and reflected images. The reflectance distributions are generated from phase differences determined by the shape of the object surface, which is constructed by using the Blinn and Torrance-Sparrow reflection models. Moreover, the reflected images are adapted when they are mapped onto metallic objects such as mirrors. Incorporating these two characteristics of reflection means that CGHs can express metallic objects realistically. Computer simulations and computational and optical reconstructed experiments were carried out. These results show the potential of the proposed method for showing metallic objects.


Impacts of drug reimbursement reductions on utilization and expenditures of oral antidiabetic medications in Taiwan: an interrupted time series study.
To control increasing pharmaceutical expenditures, Taiwan's National Health Insurance has implemented a series of drug reimbursement price reductions since 2000. This study examined changes in use and expenditures of oral antidiabetic medications following the price regulation in November 2006. We obtained claims data between January 2006 and August 2007 from Taiwan's National Health Insurance Research Database. We categorized oral antidiabetic products as affected by the reimbursement reduction ("targeted") or not ("non-targeted"), by level of relative price reduction, and by manufacturer type (international vs. local manufacturers). We used an interrupted time series design and segmented regression models to estimate changes in monthly per capita prescribing rate, volume, and insurance reimbursement expenditures following the policy. The majority (129/178; 72.5%) of oral antidiabetic products were targeted by this round of price reductions. There was a relative reduction of 9.5% [95%CI: -12.68, -6.32] in total expenditures at ten months post-policy compared to expected rates. For targeted products, there were 2.04% [95%CI: -4.15, 0.07] and 13.26% [95%CI: -16.64, -9.87] relative reductions in prescribing rate and expenditures, respectively, at ten months post-policy. Non-targeted products increased significantly (22% [95%CI: 10.49, 33.51] and 22.85% [95%CI: 11.69, 34.01] relative increases in prescribing rate and expenditures respectively). Larger reimbursement cuts led to greater reductions in prescribing rate, volume, and insurance reimbursement expenditures of targeted products. Prescribing rates of both targeted and non-targeted products by international manufacturers declined after the policy while rates of prescribing non-targeted products by local manufacturers increased. While total government expenditures for oral antidiabetic medications were contained by the policy, our results indicate that prescribing shifted at the margin from targeted to non-targeted products and from international to local products. Further research is warranted to understand how changes in medication use due to price regulation policies affect medication adherence and patient health outcomes.


Identifying problem and compulsive gamblers.
To present a meta-analysis of current research on the prevalence, identification, and treatment of problem and compulsive gamblers. Problem and compulsive gambling was not a socio-scientific concern until the last two decades. Hence research on this topic is limited. The summary and analysis for this paper relied on computer searches of journal and news abstracts in addition to direct contact with organizations addressing the identification and treatment of compulsive gamblers. An estimated 5% of those who gamble run into problems. About 1% of those who gamble are predicted to experience serious problems. Successful treatment of problem and compulsive gambling continues to be a challenge. Although cognitive therapy has been the favoured approach, a combination of several therapeutic approaches is advocated. Problem and compulsive gambling can present a real health threat. As with other addictions, treatment strategies continue to be a baffling social problem. Aware and informed physicians can have a pivotal role in the difficult process of identifying, acknowledging, and remediating problem and compulsive gambling.


Genetic Interaction Network as an Important Determinant of Gene Order in Genome Evolution.
Although it is generally accepted that eukaryotic gene order is not random, the basic principles of gene arrangement on a chromosome remain poorly understood. Here, we extended existing population genetics theories that were based on two-locus models and proposed a hypothesis that genetic interaction networks drive the evolution of eukaryotic gene order. We predicted that genes with positive epistasis would move toward each other in evolution, during which a negative correlation between epistasis and gene distance formed. We tested and confirmed our prediction with computational simulations and empirical data analyses. Importantly, we demonstrated that gene order in the budding yeast could be successfully predicted from the genetic interaction network. Taken together, our study reveals the role of the genetic interaction network in the evolution of gene order, extends our understanding of the encoding principles in genomes, and potentially offers new strategies to improve synthetic biology.


Current indications and outcomes for thoracoscopic segmentectomy for early stage lung cancer.
Management of early stage non-small cell lung cancer (NSCLC) is evolving as the epidemiology of NSCLC has changed due to decreased rates of smoking in the general population, the development of screening programs and advancements in imaging modalities. Surgical technique is also evolving as a result of changes in instrumentation. Segmentectomy for early stage NSCLC has the advantage of preservation of lung function. Video-assisted thoracic surgery (VATS) segmentectomy is a safe option for surgeons already experienced in VATS lobectomy and offers comparable oncological benefit with better quality of life in selected patients. In this article, we will examine the role of segmentectomy and VATS for early stage NSCLC.


[Stability of the structure and antigenic determinants of adenovirus type 1 native hexon to proteases].
Hexon capsomers of human adenovirus type 1 (h1) labeled by iodine 125 were digested in a native state (trimers) by trypsin, chymotrypsin or papain, and the resulting hydrolysates were analyzed by SDS-PAGE. In each case, a discrete and temporally stable pattern of relatively large fragments was revealed. The degree of hexon polypeptide hydrolysis was maximal for papain, intermediate for chymotrypsin and minimal for trypsin, the largest fragments in the digest being 32, 40 and 80 kD, respectively. At room temperature, all the electrophoretically discernible hexon proteolytical fragments were held together in structures resembling intact hexon trimers and could be regarded as "hexon cores", of which papain hexon cores were the most stable during SDS-PAGE. Radioimmunoprecipitation analysis revealed a complete absence of native hexon antigenicity in thermodenaturated fragments of hexon protease digests, while native trypsin, chymotrypsin and papain hexon cores could be precipitated by hexon-specific antibodies. The immunoprecipitated material contained all of the hexon fragments found in appropriate hexon cores and retained the structure of the original cores. Trypsin, chymotrypsin and papain hexon cores were shown to possess at least part of native Ad h1 hexon antigenic determinants of each of the following specificities: species-specific (epsilon), cross-reactive with hexon of human adenoviruses (h3 and h6), simian adenovirus (sim 16), bovine adenoviruses (bos 3 and bos 7) and avian adenovirus (Aviadenovirus gal 1 or CELO). Thus, the full spectrum of known hexon antigenic determinants (species-specific to intergenus-crossreactive) is at least portly stable against protease attack of native hexon capsomers.


Obesity diagnosis and mortality risk based on a body shape index (ABSI) and other indices and anthropometric parameters in university students
Obesity is a global epidemic and belongs to major risk factors for the most prevalent diseases. Anthropometric measures are simple, inexpensive, non-invasive tools to diagnosis obesity and to assess the risk of morbidity and mortality. The most widely used are body mass index (BMI), waist circumference (WC), waist-to-hip (WHR) and waist-to-height ratios, visceral fat area (VFA), body fat (BFP) and a new body shape index (ABSI). The aim of this study was to examine the usefulness of the ABSI in obesity diagnosis compared with other anthropometric parameters like WC, WHR, BMI, VFA, and BFP. We also compared the predictability between ABSI and above mentioned common anthropometric indices. The study group was composed of 236 university students. Body height, weight, WC was measured and BMI, WHR, ABSI and ABSI z-score were calculated. The anthropometric measurements were made by using InBody 720 (Biospace Co. Ltd., Seoul, Republic of Korea). Body composition, especially VFA, BFP, FFM was diagnosed by multifrequency bioelectrical impedance analysis. We evaluated the collected data statistically and graphically in Microsoft Office Excel 2010 (Los Angeles, CA, USA). Statistical analyses were performed using the program STATISTICA Cz version 10. The diagnosis of obesity among participants according to anthropometric measures and indices showed considerable differences. We found that obesity was diagnosed according to waist circumference in 31% of participants. According to BMI 20.3% of subjects were overweight and 5.1% obese. With increasing BMI values, the values of WC, WHR and VFA also increased linearly. According to visceral fat area 11.4% of participants were in the risk obese group and by ABSI mortality risk there were 22% of subjects with high risk (4.8% and 28.3% for men and women, respectively) and 19.1% with very high risk (11.1% and 22% for men and women, respectively). VFA and BFP values increased with increasing risk of mortality, and in men also waist circumference values. When evaluating the ABSI in relation to BMI, the U-shaped curve was confirmed and in the case of WC the J-shaped curve. The FFM evaluation showed that the very low ABSI mortality risk group reached the highest values of this parameter and the lowest values showed the average mortality risk group, not only in the study group but also in male and female groups. Our findings suggest the relevance of ABSI to screen at-risk population.


Diagnostic inadequacy of dilatation and curettage.
To assess the diagnostic inadequacy of dilatation and curettage (D&C) by comparing histologic findings with this technique with those obtained after hysterectomy. Retrospective clinical study. University-affiliated hospital. Three hundred ninety-seven patients with abnormal uterine bleeding who underwent D&C and, within 2 months, hysterectomy because of histologic findings or persistence of symptoms. Comparison of histologic findings on D&C with those obtained after hysterectomy. In 248 of 397 patients (62.5%), D&C failed to detect intrauterine disorders subsequently found at hysterectomy; the sensitivity was 46%, the specificity was 100.0%, the positive predictive value was 100.0%, and the negative predictive value was 7.1%. Dilatation and curettage is an inadequate diagnostic and therapeutic tool for all uterine disorders; this technique missed 62.5% of major intrauterine disorders, and all endometrial disorders were still present in the removed uterus.


Antifungal treatment of wild amphibian populations caused a transient reduction in the prevalence of the fungal pathogen, Batrachochytrium dendrobatidis.
Emerging infectious diseases can drive host populations to extinction and are a major driver of biodiversity loss. Controlling diseases and mitigating their impacts is therefore a priority for conservation science and practice. Chytridiomycosis is a devastating disease of amphibians that is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), and for which there is an urgent need to develop mitigation methods. We treated tadpoles of the common midwife toad (Alytes obstetricans) with antifungal agents using a capture-treat-release approach in the field. Antifungal treatment during the spring reduced the prevalence of Bd in the cohort of tadpoles that had overwintered and reduced transmission of Bd from this cohort to the uninfected young-of-the-year cohort. Unfortunately, the mitigation was only transient, and the antifungal treatment was unable to prevent the rapid spread of Bd through the young-of-the year cohort. During the winter, Bd prevalence reached 100% in both the control and treated ponds. In the following spring, no effects of treatment were detectable anymore. We conclude that the sporadic application of antifungal agents in the present study was not sufficient for the long-term and large-scale control of Bd in this amphibian system.


Controlling parental feeding practices and child body composition in ethnically and economically diverse preschool children.
Controlling parental feeding practices may be associated with childhood overweight, because coercive or intrusive feeding practices may negatively impact children's development of self-regulation of eating. This study examined pressuring or forcing a child (healthy or unhealthy foods) and restricting child from unhealthy or snack foods as two types of controlling feeding practices that explain unique variances in measures of child body composition (BMI, percent body fat, and parental perception of child weight). In an ethnically and economically diverse sample of 243 children aged 4-6years old and their biological parents (89% biological mothers, 8% biological fathers, and 3% step or grand-parent), descriptive statistics indicate ethnic and family income differences in measures of feeding practices and child body composition. Additionally, the two "objective" indices of body composition (BMI and percent body fat) were related to low pressure to eat, whereas the "subjective" index (perceived child weight) was related to restriction. Regression analyses accounting for ethnic and family income influences indicate that pressure to eat and restriction both explained unique variances in the two "objective" indices of body composition, whereas only restriction explained variance in perceived child weight. Findings have implications for helping parents learn about feeding practices that promote children's self-regulation of eating that simultaneously serves as an obesity prevention strategy.


Midbrain muscarinic receptor mechanisms underlying regulation of mesoaccumbens and nigrostriatal dopaminergic transmission in the rat.
Laterodorsal (LDT) and pedunculopontine (PPT) tegmental nuclei in the mesopontine project cholinergic inputs to the midbrain ventral tegmental area (VTA) and substantia nigra pars compacta (SNc), respectively, to directly and indirectly influence the activity of dopamine neuronal cells via actions on muscarinic and nicotinic receptors. The present study investigated the role of midbrain muscarinic receptors in the functional modulation of VTA and SNc dopamine cell activity as reflected by alterations in, respectively, nucleus accumbens (NAc) and striataldopamine efflux. In vivo chronoamperometry was used to measure changes in basal dopamine efflux via stearate-graphite paste electrodes implanted unilaterally in the NAc or striatum of urethane-anaesthetized rats, following blockade or activation of, respectively, VTA or SNc muscarinic receptors. Intra-VTA or -SNc infusion of the muscarinic antagonist scopolamine (200 microg/microL) reduced, respectively, NAc and striatal dopamine efflux while infusion of the muscarinic and nicotinic agonist carbachol (0.5 microg/microL) or the prototypical muscarinic agonist muscarine (0.5 microg/microL) increased NAc and striatal dopamine efflux. Transient decreases in dopamine efflux preceded these increases selectively in the striatum, suggesting a reduction in excitatory or increase in inhibitory drive to the SNc by preferential activation of M3 muscarinic receptors on GABA interneurons and glutamatergic inputs. This was confirmed by showing that selective blockade of M3 receptors with p-F-HHSiD (0.5 microg/microL) increased striatal, but not NAc, dopamine efflux. Together, these findings suggest that midbrain muscarinic receptors, probably M5 subtypes on VTA and SNc dopamine neurons, contribute to the tonic excitatory regulation of forebrain basal dopamine transmission whereas presynaptic M3 receptors serve to counter excessive excitation of nigral dopamine cell activity.


Is lumbar lordosis related to low back pain development during prolonged standing?
An induced-pain paradigm has been used in back-healthy people to understand risk factors for developing low back pain during prolonged standing. The purposes of this study were to (1) compare baseline lumbar lordosis in back-healthy participants who do (Pain Developers) and do not (Non-Pain Developers) develop low back pain during 2 h of standing, and (2) examine the relationship between lumbar lordosis and low back pain intensity. Cross-sectional. First, participants stood while positions of markers placed superficial to the lumbar vertebrae were recorded using a motion capture system. Following collection of marker positions, participants stood for 2 h while performing light work tasks. At baseline and every 15 min during standing, participants rated their low back pain intensity on a visual analog scale. Lumbar lordosis was calculated using marker positions collected prior to the 2 h standing period. Lumbar lordosis was compared between pain developers and non-pain developers. In pain developers, the relationship between lumbar lordosis and maximum pain was examined. There were 24 (42%) pain developers and 33 (58%) non-pain developers. Lumbar lordosis was significantly larger in pain developers compared to non-pain developers (Mean difference = 4.4°; 95% Confidence Interval = 0.9° to 7.8°, Cohen's d = 0.7). The correlation coefficient between lumbar lordosis and maximum pain was 0.46 (P = 0.02). The results suggest that standing in more lumbar lordosis may be a risk factor for low back pain development during prolonged periods of standing. Identifying risk factors for low back pain development can inform preventative and early intervention strategies.


[Metabolic changes in the patient with recurring lithiasis. Our cases].
Over a period spanning six months, 69 patients with lithiasis were selected for evaluation of mineral metabolism. Hypercalciuria, particularly renal, was the metabolic disorder that was commonly detected. A high incidence of altered uric acid metabolism was also observed. The importance of correct diagnosis and treatment of hyperparathyroidism is underscored because of the extraordinary benefit afforded to these patients. Correct patient selection has resulted in a lower incidence of those with normal metabolic parameters and has thus maximized the usefulness of these studies.