Initial staging of non-small cell lung cancer: value of routine radioisotope bone scanning.
The exclusion of bone metastases is important in the initial staging of non-small cell lung cancer, though there is debate about whether bone scans should be performed routinely or restricted to patients who present with clinical or laboratory indicators suggesting skeletal metastases. In a prospective study of 110 consecutive patients referred for initial staging of non-small cell lung cancer, we assessed the sensitivity of a group of clinical indicators (chest pain, skeletal pain, bone tenderness on physical examination, serum alkaline phosphatase, and serum calcium) for the presence of skeletal metastases as determined by bone scanning. The final staging result was validated with follow up data over at least three years. At the initial staging 37 of 110 bone scans (34%) showed areas of increased uptake, of which only nine were confirmed to be metastases (by tomography, computed tomography, or biopsy). Half the patients (55) had at least one clinical indicator suggesting skeletal metastases, including all patients with proved skeletal metastases. Thus the sensitivity of these clinical indicators was 100% and the specificity 54%. Within one year three of 27 patients with non-confirmed positive bone scans had skeletal metastases, one of which was in the area that had shown increased uptake initially. All these patients had clinical indicators for skeletal metastases and all had inoperable advanced tumours. Four of 69 patients with an initially negative bone scan developed skeletal metastases within one year. It is concluded that in non-small cell lung cancer bone scanning can be restricted to patients with clinical indicators for skeletal metastases. This approach reduces the number of bone scans and consecutive investigations without loss of sensitivity in the detection of skeletal metastases.


Optical coherence tomography in multiple sclerosis: A 3-year prospective multicenter study.
To evaluate changes over 3 years in the thickness of inner retinal layers including the peripapillary retinal nerve fiber layer (pRNFL), and combined macular ganglion cell and inner plexiform layers (mGCIPL), in individuals with relapsing-remitting multiple sclerosis (RRMS) versus healthy controls; to determine whether optical coherence tomography (OCT) is sufficiently sensitive and reproducible to detect small degrees of neuroaxonal loss over time that correlate with changes in brain volume and disability progression as measured by the Expanded Disability Status Scale (EDSS). Individuals with RRMS from 28 centers (n = 333) were matched with 64 healthy participants. OCT scans were performed on Heidelberg Spectralis machines (at baseline; 1 month; 6 months; 6-monthly thereafter). OCT measurements were highly reproducible between baseline and 1 month (intraclass correlation coefficient >0.98). Significant inner retinal layer thinning was observed in individuals with multiple sclerosis (MS) compared with controls regardless of previous MS-associated optic neuritis--group differences (95% CI) over 3 years: pRNFL: -1.86 (-2.54, -1.17) µm; mGCIPL: -2.03 (-2.78, -1.28) µm (both p < 0.0001; effect sizes 0.39 and 0.34). Greater inner retinal layer atrophy was observed in individuals diagnosed with RRMS <3 years versus >5 years (pRNFL: p < 0.05; mGCIPL: p < 0.01). Brain volume decreased by 1.3% in individuals with MS over 3 years compared to 0.5% in control subjects (effect size 0.76). mGCIPL atrophy correlated with brain atrophy (p < 0.0001). There was no correlation of OCT data with disability progression. OCT has potential to estimate rates of neurodegeneration in the retina and brain. The effect size for OCT, smaller than for magnetic resonance imaging based on Heidelberg Spectralis data acquired in this study, was increased in early disease.


[Individual medical rehabilitation for patients having occupational diseases caused by dust].
The article covers organizational and legal problems of medical rehabilitation for patients with occupational diseases caused to dust. The authors suggested steps of individual medical rehabilitation and criteria to evaluate efficiency of rehabilitation, exemplified by dust obstructive bronchitis patients.


Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population: the Tromsø study.
Ecologic and observational studies have suggested an association between serum 25-hydroxyvitamin D (25(OH)D) levels and cardiovascular disease (CVD) risk factors, CVD mortality, and cancer mortality. Based on this, low serum 25(OH)D levels should be associated with higher all-cause mortality in a general population. This hypothesis was tested in the present study. The Tromsø study is a longitudinal population-based multipurpose study initiated in 1974 with focus on lifestyle-related diseases. Our data are based on the fourth Tromsø study carried out in 1994-1995. Information about death and cause of death was registered by obtaining information from the National Directory of Residents and the Death Cause Registry. Serum 25(OH)D was measured in 7161 participants in the fourth Tromsø study. Results are presented for smokers (n=2410) and non-smokers (n=4751) separately as our immunoassay seems to overestimate 25(OH)D levels for smokers. During a mean 11.7 years of follow-up, 1359 (19.0%) participants died. In multivariate regression models, there was a significantly increased risk of all-cause mortality (hazard ratio (HR) 1.32, confidence interval (CI) 1.07-1.62) among non-smoking participants in the lowest 25(OH)D quartile when compared with participants in the highest quartile. Equivalent results for smokers were not significant (HR 1.06, CI 0.83-1.35). Low serum 25(OH)D levels were associated with increased all-cause mortality for non-smokers, but the results did not reach statistical significance for smokers. However, low 25(OH)D levels are known to be associated with impaired general health, and randomized controlled studies are needed to address the question of causality.


Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium.
Human 3-dimensional (3D) enteroid or colonoid cultures derived from crypt base stem cells are currently the most advanced ex vivo model of the intestinal epithelium. Due to their closed structures and significant supporting extracellular matrix, 3D cultures are not ideal for host-pathogen studies. Enteroids or colonoids can be grown as epithelial monolayers on permeable tissue culture membranes to allow manipulation of both luminal and basolateral cell surfaces and accompanying fluids. This enhanced luminal surface accessibility facilitates modeling bacterial-host epithelial interactions such as the mucus-degrading ability of enterohemorrhagic E. coli (EHEC) on colonic epithelium. A method for 3D culture fragmentation, monolayer seeding, and transepithelial electrical resistance (TER) measurements to monitor the progress towards confluency and differentiation are described. Colonoid monolayer differentiation yields secreted mucus that can be studied by the immunofluorescence or immunoblotting techniques. More generally, enteroid or colonoid monolayers enable a physiologically-relevant platform to evaluate specific cell populations that may be targeted by pathogenic or commensal microbiota.


Massively invasive diffuse choroidal melanoma.
A 74-year-old woman was found to have increasing proptosis in a blind, painful left eye with neovascular glaucoma. Uveal malignant melanoma with massive orbital involvement was diagnosed, and the patient underwent orbital exenteration, with preoperative and postoperative orbital irradiation. The tumor was a mixed-cell, diffuse uveal malignant melanoma with involvement of the optic nerve adjacent to the line of surgical transection and of the optic nerve sheath. Subsequently, the cerebrospinal fluid cytology disclosed cells consistent with malignant melanoma, despite the absence of neurologic signs or symptoms. Cerebrospinal fluid cytology is essential in such cases, and ultrasonography is of value.


Impact of Depression and Anxiety on Change to Physical Activity Following a Pragmatic Diabetes Prevention Program Within Primary Care: Pooled Analysis From Two Randomized Controlled Trials.
The impact of major affective disorders on the effectiveness of diabetes prevention programs at promoting health behaviors has not been established. We investigated whether depression modifies the effectiveness of two pragmatic diabetes prevention programs at promoting increased physical activity. This study pooled data from two cluster randomized controlled trials (Walking Away from Type 2 Diabetes and Let's Prevent Type 2 Diabetes) that included individuals at high risk of type 2 diabetes who were recruited from primary care. The trials used very similar intervention methods to promote physical activity and had annual follow-up over a 36-month period. Depressive symptoms were measured by the Hospital Anxiety and Depression Scale, and physical activity was measured by a piezoelectric pedometer (Let's Prevent Type 2 Diabetes) or an accelerometer (Walking Away from Type 2 Diabetes) and expressed as steps per day. This analysis included 1,163 individuals (571 control, 592 intervention) who had concurrent baseline and follow-up data for ambulatory activity, depression, and anxiety. The median depression score was 3 at baseline; 11% of individuals were classified as having mild to severe depression. Those with no depressive symptoms at baseline or during follow-up increased their ambulatory activity by 592 steps per day (P < 0.001); this effect decayed by 88 steps per day (95% CI 21, 155) for every additional depressive symptom score at baseline, and each increase in the depressive symptom score between baseline and follow-up further attenuated the intervention effect by 99 steps per day (95% CI 2, 196). Both depressive symptom burden at baseline and change in this burden are associated with a graded reduction in the effectiveness of diabetes prevention programs at increasing physical activity in primary care.


IFI16 is an essential mediator of growth inhibition, but not differentiation, induced by the leukemia inhibitory factor/JAK/STAT pathway in medullary thyroid carcinoma cells.
Activation of Ras or Raf in the human medullary thyroid carcinoma (MTC) cell line, TT, induces growth arrest and differentiation via two parallel, yet independent, pathways. One of these pathways is intracellular and the other is a cell-extrinsic, autocrine/paracrine pathway mediated by the leukemia inhibitory factor (LIF)/JAK/STAT pathway. Here, we show that IFI16 is a necessary and sufficient downstream effector for LIF effects in MTC cells, specifically required for the LIF/JAK/STAT pathway-induced growth inhibition in these cells. IFI16 was induced by Raf or LIF. Dominant-negative STAT3 could block the induction, indicating that Raf can induce IFI16 only via the cell-extrinsic pathway. Knock-down of IFI16 using siRNA abrogated LIF-induced changes in cellular levels of E2F1, cyclin D1, and p21WAF/CIP1, and cell cycle arrest. In addition, adenovirus-mediated overexpression of IFI16 was sufficient to induce growth arrest. In contrast to its essential role for LIF-mediated growth arrest, IFI16 was not required for differentiation induced by LIF. Knock-down of IFI16 could not block changes in differentiation markers of the MTC cells, including calcitonin, RET, and cell morphology. Our study identifies IFI16 as an essential growth-specific effector of the cell-extrinsic growth inhibitory pathway of Ras/Raf signaling in MTC cells.


Role of transglutaminase 2 in celiac disease pathogenesis.
A number of lines of evidence suggest that transglutaminase 2 (TG2) may be one of the earliest disease-relevant proteins to encounter immunotoxic gluten in the celiac gut. These and other investigations also suggest that the reaction catalyzed by TG2 on dietary gluten peptides is essential for the pathogenesis of celiac disease. If so, several questions are of critical significance. How is TG2 activated in the celiac gut? What are the disease-specific and general consequences of activating TG2? Can local inhibition of TG2 in the celiac intestine suppress gluten induced pathogenesis in a dose-responsive manner? And what are the long-term consequences of suppressing TG2 activity in the small intestinal mucosa? Answers to these questions will depend upon the development of judicious models and chemical tools. They also have the potential of yielding powerful next-generation drug candidates for treating this widespread but overlooked chronic disease.


A simple method to transfer, integrate and study expression of foreign genes, such as chicken ovalbumin and alpha-actin in plant tumors.
A simple method for inserting foreign genes into the T-region of Agrobacterium Ti-plasmids is described. A modified cosmid (pHC 79) was introduced into a predetermined site of the T-region of pTi C58. An Agrobacterium strain harboring this modified Ti-plasmid was used as an acceptor strain into which genes, cloned in pBR322, can be introduced by mobilization from Escherichia coli. pBR322-derived plasmids cannot replicate in Agrobacterium, but can be maintained by integration into the T-region of the modified Ti-plasmids by homologous recombination. This method was used to introduce the genes for ovalbumin and alpha-actin from chicken into tobacco tumors. Southern blotting and re-isolation of the inserted genes by reverse cloning showed that the animal DNA was transferred and integrated into the plant genome without rearrangements. The alpha-actin gene is not transcribed in plant tumors, whereas transcription of the ovalbumin gene was observed, however the initiation point of transcription was different from the one used in the chicken oviduct. The RNA transcribed from the chicken ovalbumin gene is polyadenylated and ranges in size between 2 and 7 kb.


Predictors of Change in Skin Intrinsic Fluorescence in Type 1 Diabetes: The Epidemiology of Diabetes Complications Study.
Skin intrinsic fluorescent (SIF) scores are indirect measures of advanced glycation end-products (AGEs). SIF scores are cross-sectionally associated with type 1 diabetes (T1D) complications such as increased albumin excretion rate (AER), coronary artery calcification (CAC) and neuropathy. We assessed predictors of SIF score change in those with T1D. Data from the 30-year longitudinal Epidemiology of Diabetes Complications (EDC) study of childhood-onset T1D were used to assess AGEs measured with a SIF score produced by the SCOUT DS® device. SIF scores were assessed twice in 83 participants: between 2007-08 and again between 2010-14. Regression analyses were used to assess independent predictors of SIF score change. At baseline, mean age was 47.9 ± 6.9 years, diabetes duration was 36.7 ± 6.4 years, and median glycosylated hemoglobin (HbA1c) was 7.1 (interquartile range: 6.5, 8.5). During a mean follow-up of 5.2 ± 0.9 years, mean change in SIF score was 2.9 ± 2.8 arbitrary units. In multivariable linear regression models, log HbA1c (P < 0.001), log estimated glomerular filtration rate (eGFR) (P < 0.001), overt nephropathy (defined as AER ≥ 200 µg/min, P = 0.06), and multiple daily insulin shots/pump use (MDI) exposure years (P = 0.02) were independent predictors of SIF score change. Increases in SIF score over 5 years were related to increased glycemic levels and decreased kidney function (eGFR). MDI and glomerular damage were related to a decreased SIF score. This is one of the first studies with repeated SIF assessments in T1D and provides unique, albeit preliminary, insight about these associations.


A perspective on respiratory disease in feedlot cattle.
The etiological factors causing acute pneumonia in feedlot cattle are complex in terms of their interactions with the host to cause clinical disease or death. The most significant agent is Pasteurella hemolytica, but it may be acting relatively late in the overall sequence of events.It is necessary to develop an overall strategy to determine the pathogenesis of the disease in the field. Approaches to this objective are discussed, with the emphasis on the role of the epidemiologist.


Inhibition of brain mitochondrial swelling by idebenone.
The effects of idebenone on the swelling of rat brain mitochondria were studied. When FeCl3 was added to a mitochondrial suspension, a pronounced mitochondrial swelling occurred accompanied by the production of lipid peroxide; the two phenomena were closely correlated (r = 0.96, p less than 0.01). Idebenone inhibited the mitochondrial swelling and lipid peroxidation in a concentration-dependent manner; the concentration giving 50% inhibition was 37 microM for swelling and 53 microM for lipid peroxidation. Metabolites of idebenone also inhibited the lipid peroxidation. These results suggest that idebenone stabilizes the mitochondrial membrane by inhibiting lipid peroxidation in brain mitochondria.


Hypersensitivity reactions in the small intestine. 2. Effects of allograft rejection on mucosal architecture and lymphoid cell infiltrate.
Small intestinal mucosa contains both thymus dependent and thymus independent lymphoid cells and thus has the capacity to act via humoral and cellular mechanisms as a site of local immunity and local hypersensitivity. Allograft rejection of mouse small intestine is a model of a local cell mediated reaction. The effects of this clearly defined, immunologically mediated damage villi, crypts, enterocytes, and lymphoid cell infiltrate have been assessed by comparing the morphology of rejecting allografts with that of isografts and normal small intestine of the same age. In rejection there is infiltration of the lamina propria with lymphocytes, hyperplasia of the crypts of Lieberkuhn, and an eventual sloughing off of the mucosa. Usually, but not always, there is villous atrophy and increased numbers of intraepithelial lymphocytes. However, the morphology of individual enterocytes remains normal throughout rejection and neither plasma cells nor polymorphonuclear leucocytes infiltrate the lamina propria before mucosal ulceration. These results show unequivocally that a local T cell mediated immune response causes villous atrophy and crypt hyperplasia in this animal model, and since there is no evidence of local enterocyte cytotoxicity, a lymphokine may be the link between the activated T cell and the effects on mucosal architecture. We suggest that a local CMI reaction may be the cause of villous atrophy, crypt hyperplasia, and malabsorption in many clinical and experimental conditions, including coeliac disease, food allergy, and intestinal infections.


Insights into the insect salivary gland proteome: diet-associated changes in caterpillar labial salivary proteins.
The primary function of salivary glands is fluid and protein secretion during feeding. Compared to mammalian systems, little is known about salivary protein secretion processes and the effect of diet on the salivary proteome in insect models. Therefore, the effect of diet nutritional quality on caterpillar labial salivary gland proteins was investigated using an unbiased global proteomic approach by nanoLC/ESI/tandem MS. Caterpillars of the beet armyworm, Spodoptera exigua Hübner, were fed one of three diets: an artificial diet containing their self-selected protein to carbohydrate (p:c) ratio (22p:20c), an artificial diet containing a higher nutritional content but the same p:c ratio (33p:30c) or the plant Medicago truncatula Gaertn. As expected, most identified proteins were associated with secretory processes and not influenced by diet. However, some diet-specific differences were observed. Nutrient stress-associated proteins, such as peptidyl-propyl cis-trans isomerase and glucose-regulated protein94/endoplasmin, and glyceraldehyde 3-phosphate dehydrogenase were identified in the labial salivary glands of caterpillars fed nutritionally poor diets, suggesting a link between nutritional status and vesicular exocytosis. Heat shock proteins and proteins involved in endoplasmic reticulum-associated protein degradation were also abundant in the labial salivary glands of these caterpillars. In comparison, proteins associated with development, such as arylphorin, were found in labial salivary glands of caterpillars fed 33p:30c. These results suggest that caterpillars fed balanced or nutritionally-poor diets have accelerated secretion pathways compared to those fed a protein-rich diet.


A new iterated two-band diffusion equation: theory and its application.
In this paper, we propose an iterated two-band filtering method to solve the selective image smoothing problem. We prove that a discrete computation step in an iterated nonlinear diffusion-based filtering algorithm is equivalent to a sequence of operations, including decomposition, regularization, and then reconstruction, in the proposed two-band filtering scheme. To correctly separate the high frequency components from the low frequency ones in the decomposition process, we adopt a dyadic wavelet-based approximation scheme. In the regularization process, we use a diffusivity function as a guide to retain useful data and suppress noises. Finally, the signal of the next stage, which is a "smoother" version of the signal at the previous stage, can be computed by reconstructing the decomposed low frequency component and the regularized high frequency component. Based on the proposed scheme, the smoothing operation can be applied to the correct targets. Experimental results show that our new approach is really efficient in noise removing.


Revisiting Risk Governance of GM Plants: The Need to Consider New and Emerging Gene-Editing Techniques.
New and emerging gene-editing techniques make it possible to target specific genes in species with greater speed and specificity than previously possible. Of major relevance for plant breeding, regulators and scientists are discussing how to regulate products developed using these gene-editing techniques. Such discussions include whether to adopt or adapt the current framework for GMO risk governance in evaluating the impacts of gene-edited plants, and derived products, on the environment, human and animal health and society. Product classification or definition is one of several aspects of the current framework being criticized. Further, knowledge gaps related to risk assessments of gene-edited organisms-for example of target and off-target effects of intervention in plant genomes-are also of concern. Resolving these and related aspects of the current framework will involve addressing many subjective, value-laden positions, for example how to specify protection goals through ecosystem service approaches. A process informed by responsible research and innovation practices, involving a broader community of people, organizations, experts, and interest groups, could help scientists, regulators, and other stakeholders address these complex, value-laden concerns related to gene-editing of plants with and for society.


Incidental Prostate Adenocarcinoma in Cystoprostatectomy Specimens: Partial Versus Complete Prostate Sampling.
The rate of incidental prostate adenocarcinoma (PCa) detection in radical cystoprostatectomy (RCP) varies widely, ranging from 15% to 54%. Such variability may be explained by institutional differences in prostate grossing protocols. Either partial or complete submission of the prostate gland in RCP may result in detection of clinically insignificant or significant incidental PCa. The aim of the study was to compare the clinical significance of PCa in RCP specimens in partial versus complete sampling. Seventy-two out of 158 RCP cases showed incidental PCa. The pathologic features, including Gleason score, margin status, extraprostatic extension (EPE), seminal vesicle invasion (SVI), PCa stage, and tumor volume, were assessed. The 72 cases were divided into partial (n = 21, 29.1%) and complete sampling (n = 51, 70.8%) groups. EPE was detected in 13/72 (18.1%) with 11/13 (84.6%) cases in the complete group. Positive margins were present in 11/72 (15.3%) with 9/11 (81.8%) in the complete group. SVI was detected in 4/72 (5.6%) with 3/4 (75.0%) in the complete group. Overall, 4/72 (5.6%) had a Gleason score >7, all of which were in the complete group. Our data suggest that complete sampling of the prostate may be the ideal approach to grossing RCP specimens, allowing for greater detection of clinically significant incidental PCa.


RNA-Catalyzed Polymerization of Deoxyribose, Threose, and Arabinose Nucleic Acids.
An RNA-dependent RNA polymerase ribozyme that was highly optimized through in vitro evolution for the ability to copy a broad range of template sequences exhibits promiscuity toward other nucleic acids and nucleic acid analogues, including DNA, threose nucleic acid (TNA), and arabinose nucleic acid (ANA). By operating on various RNA templates, the ribozyme catalyzes multiple successive additions of DNA, TNA, or ANA monomers, although with reduced efficiency compared to RNA monomers. The ribozyme can also copy DNA or TNA templates to complementary RNAs, and to a lesser extent it can operate when both the template and product strands are composed of DNA, TNA, or ANA. These results suggest that polymerase ribozymes, which are thought to have replicated RNA genomes during the early history of life, could have transferred RNA-based genetic information to and from DNA, enabling the emergence of DNA genomes prior to the emergence of proteins. In addition, genetic systems based on nucleic acid-like molecules, which have been proposed as precursors or contemporaries of RNA-based life, could have been operated upon by a promiscuous polymerase ribozyme, thus enabling the evolutionary transition between early genetic systems.


Recursive least-squares backpropagation algorithm for stop-and-go decision-directed blind equalization.
Stop-and-go decision-directed (S-and-G-DD) equalization is the most primitive blind equalization (BE) method for the cancelling of intersymbol-interference in data communication systems. Recently, this scheme has been applied to complex-valued multilayer feedforward neural network, giving robust results with a lower mean-square error at the expense of slow convergence. To overcome this problem, in this work, a fast converging recursive least squares (RLS)-based complex-valued backpropagation learning algorithm is derived for S-and-G-DD blind equalization. Simulation results show the effectiveness of the proposed algorithm in terms of initial convergence.


Polygenic risk scores as a marker for epilepsy risk across lifetime and after unspecified seizure events.
A diagnosis of epilepsy has significant consequences for an individual but is often challenging in clinical practice. Novel biomarkers are thus greatly needed. Here, we investigated how common genetic factors (epilepsy polygenic risk scores, [PRSs]) influence epilepsy risk in detailed longitudinal electronic health records (EHRs) of > 360k Finns spanning up to 50 years of individuals' lifetimes. Individuals with a high genetic generalized epilepsy PRS (PRS GGE ) in FinnGen had an increased risk for genetic generalized epilepsy (GGE) (hazard ratio [HR] 1.55 per PRS GGE standard deviation [SD]) across their lifetime and after unspecified seizure events. Effect sizes of epilepsy PRSs were comparable to effect sizes in clinically curated data supporting our EHR-derived epilepsy diagnoses. Within 10 years after an unspecified seizure, the GGE rate was 37% when PRS GGE > 2 SD compared to 5.6% when PRS GGE < -2 SD. The effect of PRS GGE was even larger on GGE subtypes of idiopathic generalized epilepsy (IGE) (HR 2.1 per SD PRS GGE ). We further report significantly larger effects of PRS GGE on epilepsy in females and in younger age groups. Analogously, we found significant but more modest focal epilepsy PRS burden associated with non-acquired focal epilepsy (NAFE). We found PRS GGE specifically associated with GGE in comparison with >2000 independent diseases while PRS NAFE was also associated with other diseases than NAFE such as back pain. Here, we show that epilepsy specific PRSs have good discriminative ability after a first seizure event i.e. in circumstances where the prior probability of epilepsy is high outlining a potential to serve as biomarkers for an epilepsy diagnosis.


Changes in mosquito species and blood meal composition associated with adulticide applications.
Although adulticide application is a pillar in the integrated management of nuisance and vector mosquitoes, non-target effects of adulticide applications within ecosystems are a substantial concern. However, the impacts of adulticide applications on non-target organisms are not necessarily detrimental, and in some cases, may provide benefits to certain organisms or wildlife. Here, we hypothesized that adulticide applications have beneficial non-target impacts on vertebrate wildlife through reduced biting pressure. To test this, we collected mosquitoes from ultra-low volume Permanone-treated (intervention) and untreated (reference) areas and assessed mosquito abundance and diversity, and abundance of blood-engorged female mosquitoes. We performed DNA barcoding analysis on mosquito blood meals to identify host species. Our results demonstrated a significant reduction in mosquito abundance by 58.9% in the intervention areas, taking into account the reduction in reference areas. Consequently, this decline led to a 64.5% reduction in the abundance of blood-engorged females. We also found a temporal dynamic of mosquito composition driven by mosquito control actions in which different mosquito species became dominant at treated sites while composition at reference areas remained similar during the same period. The present study suggests that the beneficial effects of mosquito control treatments for humans extend to other vertebrates, which represents an unstudied and rarely recognized non-target impact.


Progress and challenges in predicting protein methylation sites.
Protein methylation catalyzed by methyltransferases carries many important biological functions. Methylation and their regulatory enzymes are involved in a variety of human disease states, raising the possibility that abnormally methylated proteins can be disease markers and methyltransferases are potential therapeutic targets. Identification of methylation sites is a prerequisite for decoding methylation regulatory networks in living cells and understanding their physiological roles that have been implicated in the pathological processes. Due to various limitations of experimental methods, in silico approaches for identifying novel methylation sites have become increasingly popular. In this review, we summarize the progress in the prediction of protein methylation sites from the dataset, feature representation, prediction algorithm and online resources in the past ten years. We also discuss the challenges that are faced while developing novel predictors in the future. The development and application of methylation site prediction is a promising field of systematic biology, provided that protein methyltransferases, species and functional information will be taken into account.


Hydroxyl radical generation by red tide algae.
The unicellular marine phytoplankton Chattonella marina is known to have toxic effects against various living marine organisms, especially fishes. However, details of the mechanism of the toxicity of this plankton remain obscure. Here we demonstrate the generation of superoxide and hydroxyl radicals from a red tide unicellular organism, C. marina, by using ESR spectroscopy with the spin traps 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and N-t-butyl-alpha-phenylnitrone (PBN), and by using the luminol-enhanced chemiluminescence response. The spin-trapping assay revealed productions of spin adduct of superoxide anion (O2-) (DMPO-OOH) and that of hydroxyl radical (.OH) (DMPO-OH) in the algal suspension, which was not observed in the ultrasonic-ruptured suspension. The addition of superoxide dismutase (500 U/ml) almost completely inhibited the formation of both DMPO-OOH and DMPO-OH, and carbon-centered radicals were generated with the disappearance of DMPO-OH after addition of 5% dimethyl sulfoxide (Me2SO) and 5% ethanol. Furthermore, the generation of methyl and methoxyl radicals, which are thought to be produced by the reaction of hydroxyl radical and Me2SO under aerobic condition, was identified using spin trapping with a combination of PBN and Me2SO. Luminol-enhanced chemiluminescence assay also supported the above observations. These results clearly indicate that C. marina generates and releases the superoxide radical followed by the production of hydroxyl radical to the surrounding environment. The velocity of superoxide generation by C. marina was about 100 times faster than that by mammalian phagocytes per cell basis. The generation of oxygen radical is suggested to be a pathogenic principle in the toxication of red tide to susceptible aquaculture fishes and may be directly correlated with the coastal pollution by red tide.


Changing epidemiology of malignant cutaneous melanoma in Europe 1953-1997: rising trends in incidence and mortality but recent stabilizations in western Europe and decreases in Scandinavia.
We analyzed time trends in incidence of and mortality from malignant cutaneous melanoma in European populations since 1953. Data were extracted from the EUROCIM database of incidence data from 165 cancer registries. Mortality data were derived from the WHO database. During the 1990s, incidence rates were by far highest in northern and western Europe, whereas mortality was higher in males in eastern and southern Europe. Melanoma rates have been rising steadily, albeit with substantial geographic variation. In northern Europe, a deceleration in these trends occurred recently in persons aged under 70. Joinpoint analyses indicated that changes in these trends took place in the early 1980s. In western Europe, mortality rates have also recently leveled off [estimated annual percentage change (EAPC) from -13.6% (n.s.) to 3.3%], whereas in eastern and southern Europe both incidence and mortality rates are still increasing [incidence EAPCs 2.3-8.9%, mortality EAPCs -1.8% (n.s.) to 7.2%]. Models including the effects of age, period and birth cohort were required to adequately describe the rising incidence trends in most European populations, with a few exceptions. Time trends in mortality were adequately summarized on fitting either an age-cohort model (with the leveling off of rates starting in birth cohorts between 1930 and 1940) or an age-period-cohort model. The most plausible explanations for the deceleration or decline in the incidence and mortality trends in recent years in northern (and to a lesser extent western) Europe are earlier detection and more frequent excision of pigmented lesions and a growing public awareness of the dangers of excessive sunbathing.


Milk thistle seed extract protects rat C6 astroglial cells from acute cocaine toxicity.
Cocaine is a powerful addictive drug, widely abused in most Western countries. It easily reaches various domains within and outside of the central nervous system (CNS), and triggers varying levels of cellular toxicity. No pharmacological treatment is available to alleviate cocaine-induced toxicity in the cells without side-effects. Here, we discerned the role of milk thistle (MT) seed extract against cocaine toxicity. First, we investigated acute cytotoxicity induced by treatment with 2, 3 and 4 mM cocaine for 1 h in astroglial, liver and kidney cells in vitro, and then in living shrimp larvae in vivo. We showed that astroglial cells are more sensitive to cocaine than liver, kidney cells or larvae. Cocaine exposure disrupted the general architecture of astroglial cells, induced vacuolation, decreased cell viability, and depleted the glutathione (GSH) level. These changes may represent the underlying pathology of cocaine in the astrocytes. By contrast, MT pretreatment (200 µg/ml) for 30 min sustained the cell morphological features and increased both cell viability and the GSH level. Besides its protective effects, the MT extract was revealed to be non-toxic to astroglial cells, and displayed high free-radical scavenging activity. The results from this study suggest that enhanced GSH level underlies cell protection, and indicate that compounds that promote GSH synthesis in the cells may be beneficial against cocaine toxicity.


Techniques for the Assessment of In Vitro and In Vivo Antifungal Combinations.
Systemic fungal infections are associated with high mortality rates despite adequate treatment. Moreover, acquired resistance to antifungals is increasing, which further complicates the therapeutic management. One strategy to overcome antifungal resistance is to use antifungal combinations. In vitro, several techniques are used to assess drug interactions, such as the broth microdilution checkerboard, agar-diffusion methods, and time-kill curves. Currently, the most widely used technique is the checkerboard method. The aim of all these techniques is to determine if the interaction between antifungal agents is synergistic, indifferent, or antagonistic. However, the interpretation of the results remains difficult. Several methods of analysis can be used, based on different theories. The most commonly used method is the calculation of the fractional inhibitory concentration index. Determination of the usefulness of combination treatments in patients needs well-conducted clinical trials, which are difficult. It is therefore important to study antifungal combinations in vivo, in experimental animal models of fungal infections. Although mammalian models have mostly been used, new alternative animal models in invertebrates look promising. To evaluate the antifungal efficacy, the most commonly used criteria are the mortality rate and the fungal load in the target organs.


Surgical ward round quality and impact on variable patient outcomes.
To investigate the relationship between variability in surgical ward round (WR) quality and clinical outcomes. Evidence increasingly suggests that ward-based care plays a key role in surgical outcomes. The WR is the focal point of surgical inpatient care. Assimilating various sources of clinical information is necessary for thorough patient assessment during the WR; whether this relates to outcomes has not previously been examined. WRs were observed for patients on a surgical high-dependency unit in a tertiary academic surgical unit. All sources of clinical information (SCI) were considered. Thoroughness of assessment, defined as the percentage of SCI assessed by the clinician, was recorded as a marker of WR quality. Complications were recorded from patient records; preventability was based on Agency for Healthcare and Research Quality guidelines. The relationship between WR quality and incidence of preventable complications was analyzed. Sixty-nine WRs were observed over 37 days for 50 patients receiving care in the high-dependency unit. Observed morbidity rate was 60% (30/50). Seventy-four percent of all complications (35/46) occurred on the high-dependency unit. There was significant variability in WR quality: clinicians assessed 9% to 91% (mean = 55% ± 17%) of SCI (analysis of variance P = 0.025). Low-quality (% SCI assessed less than the mean) WRs resulted in a greater incidence of patients experiencing preventable complications [83% (10/12) vs 39% (7/18)] (P = 0.034), odds ratio = 6.43 (95% confidence interval = 1.05-39.3). Forty-one percent of complications (19/46) could have been diagnosed earlier or possibly prevented. Patient assessment during WRs is variable. Less thorough WRs result in delayed diagnoses and preventable complications, and they negatively affect outcomes. Focusing on WR quality and training may improve patient care.


ABC transporters in the liver fluke Opisthorchis felineus.
ATP-binding cassette transporters (ABC transporters) are essential components of normal cellular physiological machinery in all eukaryotic and prokaryotic species, including parasites. Some ABC transporters, e.g., P-glycoproteins, are involved in the efflux of toxins and xenobiotics from the cell. At present, nothing is known about ABC transporter genes in epidemiologically important liver flukes from the Opisthorchiidae family, including European liver fluke Opisthorchis felineus. Opisthorchiasis caused by O. felineus is a serious public health problem on the territory of Russia and other Eastern European countries. ABC drug transporters are attractive objects of research on molecular markers of resistance and on ways to potentiate sensitivity to anthelmintics through suppression of the transporters themselves with specific inhibitors. Here we aimed at the identification of ABC transporters in the O. felineus transcriptome and identification of P-glycoproteins. In addition, our aim was to assess ABC transcript abundance in the RNA-seq data, to study the mRNA expression of P-glycoprotein genes by Droplet Digital PCR throughout the life cycle of O. felineus, and to test the gene induction in response to xenobiotics or anthelminthic agents. We found 23 nucleotide sequences encoding ABC transporters belonging to different subfamilies, including four sequences of P-glycoproteins. According to the transcript abundance in the RNA-seq data, one of P-glycoproteins (P4) has the highest expression among all ABC genes in the adult worm. P-glycoproteins showed substantially differential mRNA expression throughout the fluke life cycle, with high expression in the adult worms. Putative activity of P-glycoproteins as xenobiotic efflux pumps was found to be linked to the excretory system of O. felineus and to be inhibited by verapamil or tariquidar. Thus, ABC drug transporters in the liver fluke O. felineus are functionally active, indicating that ABC drug transporters are likely to be molecular targets for a combination therapy aimed at prevention of a xenobiotic removal from helminth tissues and at increasing the drug concentration in the tissues.


Are Nucleated Cell Counts Useful in the Diagnosis of Infection in Periprosthetic Fracture?
Evaluating for the possibility of prosthetic joint infection in the setting of periprosthetic fracture is important because it determines the course of treatment. However, fracture-related inflammation can make investigations used in the diagnosis of infection less reliable. The purpose of our study was to evaluate synovial fluid nucleated cell counts as a diagnostic test for deep prosthetic infection in patients with periprosthetic fractures around hip and knee arthroplasties. Specifically, we wished to determine the test's properties (sensitivity, specificity, positive predictive value [PPV], and negative predictive value [NPV]) using threshold levels of nucleated cell counts as they are otherwise used in the diagnosis of periprosthetic infection. Billing codes were used to identify all cases of revision total hip arthroplasty (THA), revision total knee arthroplasty (TKA), open reduction and internal fixation (ORIF) of the femur, and ORIF of the tibia at our institution between 2005 and 2013. A total of 2537 charts were identified and reviewed to reveal 269 patients with 269 periprosthetic fractures about a THA or TKA (10.6% of charts reviewed). Of these, 27 fractures in 27 patients (10% of the periprosthetic fractures identified) underwent aspiration of their total joint arthroplasty to rule out infection before surgical intervention. The decision to aspirate was made by the treating surgeon based on clinical suspicion of infection from the patient history, physical examination, and radiographic findings. Nucleated cell counts from joint aspirates were recorded for all 27 patients. Synovial fluid culture results were then used to calculate the sensitivity, specificity, PPV, and NPV of an elevated nucleated cell count in the diagnosis of infection. The specificity, sensitivity, PPV, and NPV of an elevated nucleated cell count in the diagnosis of infection were 64% (95% confidence interval [CI, 34.94-75.57]), 100% (95% CI, 19.29-100), 18% (95% CI, 2.37-45.46), and 100% (95% CI, 76.66-100), respectively. Eleven of 27 patients (41%) with joint aspirates had elevated nucleated cell counts. Only two of the 11 patients (18%) with elevated nucleated cell counts had positive synovial fluid cultures. None of the patients with normal nucleated cell counts had positive synovial fluid cultures. Although quite common, an elevated nucleated cell count has moderate specificity and poor PPV in the diagnosis of infection in the setting of periprosthetic fracture. Level IV, therapeutic study.


Diacylglycerol kinase inhibitor R59022 and stimulated neutrophil responses.
The generation of phosphatidic acid in neutrophils stimulated by the chemotactic factor formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe) is inhibited by the diacylglycerol kinase inhibitor R59022. Superoxide generation produced by fMet-Leu-Phe, leukotriene B4, platelet-activating factor, or phorbol 12-myristate 13-acetate can be greatly increased in neutrophils pretreated with R59022. The potentiation occurs in the presence or absence of cytochalasin B and is evident in the absence of extracellular calcium. In addition, where the superoxide generated by fMet-Leu-Phe is not inhibited by the protein kinase C inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), the increase by R59022 is diminished by this compound. Unlike cytochalasin B, R59022 does not affect the increase in cytoskeletal actin produced by fMet-Leu-Phe or platelet-activating factor nor does it decrease the basal level. Furthermore, the basal intracellular concentration of free calcium, but not the rise produced by fMet-Leu-Phe or platelet-activating factor, is elevated by R59022. The data presented here suggest that the potentiation by R59022 of the oxidative burst is most likely mediated through protein kinase C.


Vitellogenesis and the vitellogenin gene family.
Vitellogenin is synthesized under estrogen control in the liver, extensively modified, transported to the ovary, and there processed to the yolk proteins lipovitellin and phosvitin. In the frog Xenopus laevis there are at least four distinct but related vitellogenin genes. The two genes A1 and A2 have a 95 percent sequence homology in their messenger RNA coding regions, and contain 33 introns that interrupt the coding region (exons) at homologous positions. Sequences and lengths of analogous introns differ, and many introns contain repetitive DNA elements. The introns in these two genes that have apparently arisen by duplication have diverged extensively by events that include deletions, insertions, and probably duplications. Rapid evolutionary change involving rearrangements and the presence of repeated DNA suggests that the bulk of the sequences within introns may not have any specific function.


Voltage- and time-dependent block of iK1 underlying Ba2+-induced ventricular automaticity.
The mechanism underlying the Ba2+-induced automaticity was studied in isolated guinea pig ventricular myocytes using the whole-cell clamp method and a patch electrode. In the presence of 0.1-0.3 mM Ba2+, application of a weak depolarizing current induced repetitive firing of spontaneous action potentials. Application of tetrodotoxin or Ca2+ channel blockers and removal of external Na+ and Ca2+ did not abolish the rhythmic activity, thereby suggesting that activation of inward currents played no crucial role in the generation of this rhythmicity. Voltage-clamp studies revealed that Ba2+ blocked the inward rectifier K+ current (iK1) in a voltage- and time-dependent manner with a greater and more rapid block at more negative potentials. This Ba2+ action could quantitatively be fitted with a model of the conventional bimolecular adsorption isotherm with a voltage-dependent dissociation constant. Simulation studies using this model showed that a membrane model, in which only the iK1 system and a small leak conductance were incorporated, could reproduce an automatic activity similar to that seen experimentally. Thus, in isolated ventricular cells, the voltage- and time-dependent block of iK1 by Ba2+ appears to be one important mechanism underlying the automaticity.


Growth cone collapse induced by semaphorin 3A requires 12/15-lipoxygenase.
Detection of a repellent factor, such as a semaphorin (Sema), causes localized collapse of the growth cone and directs the neurite away from the repellent. Growth cone collapse results from concomitant cytoskeletal rearrangements and detachment of adhesion sites from the extracellular matrix, via mostly unknown signaling mechanisms. In cultures of dorsal root ganglion neurons, we found that Sema3A treatment stimulates the synthesis of the eicosanoid, 12(S)-hydroxyeicosatetraenoic acid (HETE), whereas Sema3A-induced growth cone collapse is prevented when 12(S)-HETE synthesis is blocked with an inhibitor of 12/15-lipoxygenase (LO). Exogenously applied product of 12/15-LO, 12(S)-HETE, mimics Sema3A-induced collapse. As observed by interference reflection and confocal microscopy, 12(S)-HETE causes the loss of growth cone adhesion sites. The adhesion site effect seems partially independent of the actin cytoskeleton because growth cones treated with Sema3A and 12/15-LO inhibitor remain spread despite actin cytoskeleton loss. These studies demonstrate that 12/15-LO activity is a necessary step in Sema3A collapse signaling in growth cones and suggest a mechanism for its action.


Association of microRNA-7 and its binding partner CDR1-AS with the prognosis and prediction of 1st-line tamoxifen therapy in breast cancer.
The large number of non-coding RNAs (ncRNAs) and their breadth of functionalities has fuelled many studies on their roles in cancer. We previously linked four microRNAs to breast cancer prognosis. One of these microRNAs, hsa-miR-7, was found to be regulated by another type of ncRNA, the circular non-coding RNA (circRNA) CDR1-AS, which contains multiple hsa-miR-7 binding sites. Based on this finding, we studied the potential clinical value of this circRNA on breast cancer prognosis in a cohort based on a cohort that was previously analysed for hsa-miR-7 and in an adjuvant hormone-naïve cohort for 1st-line tamoxifen treatment outcomes, in which we also analysed hsa-miR-7. A negative correlation was observed between hsa-miR-7 and CDR1-AS in both cohorts. Despite associations with various clinical metrics (e.g., tumour grade, tumour size, and relapse location), CDR1-AS was neither prognostic nor predictive of relevant outcomes in our cohorts. However, we did observe stromal CDR1-AS expression, suggesting a possible cell-type specific interaction. Next to the known association of hsa-miR-7 expression with poor prognosis in primary breast cancer, we found that high hsa-miR-7 expression was predictive of an adverse response to tamoxifen therapy and poor progression-free and post-relapse overall survival in patients with recurrent disease.


Practical Considerations for Antibodies in Myeloma.
The development of the monoclonal antibodies daratumumab and elotuzumab has expanded treatment options for multiple myeloma and led to great improvement in patient outcomes. These agents have favorable safety profiles and synergize effectively with established agents used in the management of myeloma, namely immunomodulatory drugs and proteasome inhibitors. This article reviews the rationale for use of monoclonal antibodies in myeloma, current approved indications for daratumumab and elotuzumab, the manner in which these agents are used in the overall management of myeloma, and specific challenges associated with their use in the clinic. It also highlights other, emerging drug combinations that incorporate daratumumab or elotuzumab and profiles new therapeutic antibodies currently under development.


Differential induction of nuclear factor-kappaB and activator protein-1 activity after CD40 ligation is associated with primary human hepatocyte apoptosis or intrahepatic endothelial cell proliferation.
CD40, a tumor necrosis factor receptor superfamily member, is up-regulated on intraheptatic endothelial cells (IHEC) and epithelial cells during inflammatory liver disease, and there is evidence that the functional outcome of CD40 ligation differs between cell types. Ligation of CD40 on cholangiocytes or hepatocytes results in induction of Fas-mediated apoptosis, whereas ligation of IHEC CD40 leads to enhanced chemokine secretion and adhesion molecule expression. We now report that differential activation of two transcription factors, nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), in primary human hepatocytes or IHEC, is associated with and may explain, in part, the different responses of these cell types to CD40 ligation. CD40 ligation induced a rise in NF-kappaB activity in hepatocytes,which peaked at 2 h and returned to baseline by 24 h; however, IHEC CD40 ligation resulted in a sustained up-regulation of NF-kappaB (>24 h). In hepatocytes, CD40 ligation led to sustained up-regulation of AP-1 activity >24 h associated with increased protein levels of RelA (p65), c-Jun, and c-Fos, whereas no induction of AP-1 activity was observed in IHECs. Analysis of mitogen-activated protein kinase phosphorylation (phospho-extracellular signal-regulated kinase 1/2 and phospho-c-Jun NH(2)-terminal kinase 1/2) and expression of inhibitor kappaBalpha were entirely consistent, and thus confirmed the profiles of NF-kappaB and AP-1 signaling and the effects of the selective inhibitors assessed using electrophoretic mobility shift assay or Western immunoblotting. CD40 ligation resulted in induction of apoptosis in hepatocytes after 24 h, but on IHECs, CD40 ligation resulted in proliferation. Inhibition of (CD40-mediated) NF-kappaB activation prevented IHEC proliferation and led to induction of apoptosis. Selective extracellular signal-regulated kinase and c-Jun NH(2)-terminal kinase inhibitors reduced levels of apoptosis in (CD40-stimulated) hepatocytes by approximately 50%. We conclude that differential activation of these two transcription factors in response to CD40 ligation is associated with differences in cell fate. Transient activation of NF-kappaB and sustained AP-1 activation is associated with apoptosis in hepatocytes, whereas prolonged NF-kappaB activation and a lack of AP-1 activation in IHECs result in proliferation.


Effect of intramuscular polysulfated glycosaminoglycans on bovine semen production and quality of the cryopreserved semen.
Polysulfated glycosaminoglycans (PSGAG) are widely used to treat cartilage disease in horses. The drug's safety in breeding animals has not been documented. This study determined whether im PSGAG is detrimental to the bull's health and, in particular, the quantity of semen produced or the quality of the cryopreserved product. No clinically significant effect of treatment on hematologic or serum biochemistry parameters was detected, but a significant treatment x time interaction for mean corpuscular volume warrants further investigation. There was an unexplained treatment effect on total daily sperm numbers collected, even though the pre-treatment values were included as co-variates in the statistical analysis to control for any day 0 variability. There was no effect on the percent forward motile sperm in the fresh ejaculate. Although there was no significant effect of treatment on the post-thaw semen data, the power of each statistical analysis was low, ranging from 10% to 19%. Further studies using larger sample sizes are warranted. These preliminary studies suggest that im PSGAG is safe to use in bulls on an experimental basis.


Protection of human corneal epithelial cells from TNF-α-induced disruption of barrier function by rebamipide.
TNF-α disrupts the barrier function of cultured human corneal epithelial (HCE) cells. We investigated the effects of the cytoprotective drug rebamipide on this barrier disruption by TNF-α as well as on corneal epithelial damage in a rat model of dry eye. The barrier function of HCE cells was evaluated by measurement of transepithelial electrical resistance. The distribution of tight-junction (ZO-1, occludin) and adherens-junction (E-cadherin, β-catenin) proteins, and the p65 subunit of nuclear factor-κB (NF-κB) was determined by immunofluorescence microscopy. Expression of junctional proteins as well as phosphorylation of the NF-κB inhibitor IκB-α and myosin light chain (MLC) were examined by immunoblot analysis. A rat model of dry eye was developed by surgical removal of exorbital lacrimal glands. Rebamipide inhibited the disruption of barrier function as well as the downregulation of ZO-1 expression, and the disappearance of ZO-1 from the interfaces of neighboring HCE cells induced by TNF-α. It also inhibited the phosphorylation and downregulation of IκB-α, the translocation of p65 to the nucleus, the formation of actin stress fibers, and the phosphorylation of MLC induced by TNF-α in HCE cells. Treatment with rebamipide eyedrops promoted the healing of corneal epithelial defects as well as attenuated the loss of ZO-1 from the surface of corneal epithelial cells in rats. Rebamipide protects corneal epithelial cells from the TNF-α-induced disruption of barrier function by maintaining the distribution and expression of ZO-1 as well as the organization of the actin cytoskeleton. Rebamipide is, thus, a potential drug for preventing or ameliorating the loss of corneal epithelial barrier function associated with ocular inflammation.


Mild cognitive impairment in Parkinson's disease versus Alzheimer's disease.
No known studies have compared longitudinal characteristics between individuals with incident mild cognitive impairment due to Parkinson's disease (PD-MCI) versus Alzheimer's Disease (AD-MCI). We used longitudinal data from the National Alzheimer's Coordinating Center's Uniform Data Set to compare 41 PD-MCI and 191 AD-MCI participants according to their demographics, presence of ≥1 APOE e4 allele, and baseline and change over time in clinical characteristics, neuropsychological test scores, and Clinical Dementia Rating sum of boxes (CDR-SB). Multivariable linear regression models with generalized estimating equations were used to account for clustered data and to test for baseline and longitudinal differences in neuropsychological test scores. PD-MCI and AD-MCI participants differed by many demographic and clinical characteristics. Significantly fewer PD-MCI participants developed dementia over one year. Compared to AD-MCI participants, PD-MCI participants performed better at baseline and over time on a global measure of cognition (Mini Mental State Exam), memory measures (immediate and delayed Logical Memory), and a language measure (Boston Naming Test), and additionally performed better over time on an attention measure (Digit Span Forward), a language measure (Vegetable List), a processing speed measure (Digit Symbol), and an overall measure of memory and functional impairment (CDR-SB). Our study provides further evidence that PD-MCI is clinically distinct from AD-MCI and requires different tools for diagnosis and monitoring clinical progression. More importantly, this study suggests that PD-MCI takes longer to convert into dementia than AD-MCI, findings that require replication by other studies.


[Establishment of drug screening assay and pharmacodynamic evaluation method targeting influenza RNA polymerase].
Influenza virus RNA-dependent RNA polymerase (RdRP) is essential for replication and expression of influenza virus genome. Viral genomic sequences encoding RdRP are highly conservative, thus making it a potential anti-influenza drug target. A cell-based influenza RdRP inhibitor screening assay was established by a luciferase reporter system to analyze the activity of RdRP. Specificity study and statistic analysis showed that the screening assay is sensitive and reproducible.


Effects of ethanol on the intraovarian insulin-like growth factor-1 system in the prepubertal rat.
Insulin-like growth factor-1 (IGF-1) is considered to play an important role during ovarian development and function. Because ethanol (ETOH) is a gonadal toxin in men, as well as male and female rats, we hypothesized that this drug may be having detrimental effects in the ovary by altering the intraovarian actions of IGF-1. In support of this notion, the present study was undertaken to examine the chronic effects of ETOH on the ovarian IGF-1 system in prepubertal female rats. Each rat was implanted with a gastric cannula on day 24 and began receiving either a control or ETOH liquid diet on day 29. The animals were killed on day 34, confirmed to be in the late juvenile stage of development, and their ovaries and blood were collected. Using an RNase protection assay, we determined the expression of mRNAs encoding IGF-1 and the Type 1 IGF receptor in the ovaries of control and ETOH-treated rats. Results indicate that the ETOH-treated rats showed an increase in the ovarian expression of IGF-1a (p < 0.0001) and IGF-1b (p < 0.001) mRNA, the two alternatively spliced forms of the IGF-1 gene. Conversely, ovarian IGF-1 protein levels were depressed (p < 0.05) in ETOH-treated rats as determined by radioimmunoassay. Furthermore, ETOH-treated rats showed a decrease (p < 0.01) in the expression of Type-1 IGF receptor mRNA with a subsequent decrease (p < 0.05) in the ovarian levels of IGF-1 receptor protein, as determined by Western blot analysis. Also, using Western immunoblotting, we determined increases in immunoreactive IGF-binding proteins-3 (p < 0.05) and 5 (p < 0.01), but not 4, in ETOH-treated rats as compared with controls. Furthermore, we observed a concomitant decrease (p < 0.01) in the serum levels of estradiol. These results demonstrate for the first time that chronic ETOH administration is capable of altering the prepubertal intraovarian IGF-1 signaling system. We suggest that, at least in part, these effects contribute to altered prepubertal ovarian function after chronic exposure to ETOH.


Calcium, magnesium, and psychotic symptoms in a girl with idiopathic hypoparathyroidism.
A case of psychosis due to idiopathic hypoparathyroidism (IHP) has been followed for over 9 years. The optimal treatment for IHP psychosis was found to be a combination of ergocalciferol and fairly large doses of antipsychotic medication. In general, psychotic episodes occurred when there was hypocalcemia, hypercalcemia, or hypomagnesemia. Antipsychotic medication was not efficacious unless serum calcium and magnesium levels were both normal. It is suggested that psychiatric disturbances in hypoparathyroidism and treatment resistance to antipsychotic medication in other functional psychoses may also be related to hypomagnesemia.


Semipolar InGaN quantum-well laser diode with integrated amplifier for visible light communications.
GaN-based semiconductor optical amplifier (SOA) and its integration with laser diode (LD) is an essential building block yet to be demonstrated for III-nitride photonic integrated circuits (PICs) at visible wavelength. This paper presents the InGaN/GaN quantum well (QW) based dual-section LD consisting of integrated amplifier and laser gain regions fabricated on a semipolar GaN substrate. The threshold current in the laser gain region was favorably reduced from 229mA to 135mA at SOA driving voltages, VSOA, of 0V and 6.25V, respectively. The amplification effect was measured based on a large gain of 5.7 dB at VSOA = 6.25V from the increased optical output power of 8.2 mW to 30.5 mW. Such integrated amplifier can be modulated to achieve Gbps data communication using on-off keying technique. The monolithically integrated amplifier-LD paves the way towards the III-nitride on-chip photonic system, providing a compact, low-cost, and multi-functional solution for applications such as smart lighting and visible light communications.


Diuretic state affects ascending thin limb tight junctions.
The nephron segments in the inner medulla are part of the urine concentrating mechanism. Depending on the diuretic state, they are facing a large range of extracellular osmolality. We investigated whether water homeostasis affects tubular transport and permeability properties in inner medullary descending thin limb (IMdTL) and ascending thin limb (IMaTL). Three experimental groups of rats under different diuretic states were investigated on metabolic cages: waterload, furosemide-induced diuresis, and control (antidiuresis). Urine production and osmolalities reflected the 3-day treatment. To functionally investigate tubular epithelial properties, we performed experiments in freshly isolated inner medullary thin limbs from these animals. Tubular segments were acutely dissected and investigated for trans- and paracellular properties by in vitro perfusion and electrophysiological analysis. IMdTL and IMaTL were distinguished by morphological criteria. We confirmed absence of transepithelial electrogenic transport in thin limbs. Although diffusion potential measurements showed no differences between treatments in IMdTLs, we observed increased paracellular cation selectivity under waterload in IMaTLs. NaCl diffusion potential was -5.64 ± 1.93 mV under waterload, -1.99 ± 1.72 mV under furosemide-induced diuresis, and 0.27 ± 0.40 mV under control. The corresponding permeability ratio PNa/Cl was 1.53 ± 0.21 (waterload), 1.22 ± 0.18 (furosemide-induced diuresis), and 0.99 ± 0.02 (control), respectively. Claudins are main constituents of the tight junction responsible for paracellular selectivity; however, immunofluorescence did not show qualitative differences in claudin 4, 10, and 16 localization. Our results show that IMaTLs change tight junction properties in response to diuretic state to allow adaptation of NaCl reabsorption.


Transcripts from a novel BMPR2 termination mutation escape nonsense mediated decay by downstream translation re-initiation: implications for treating pulmonary hypertension.
Bone morphogenetic protein receptor type 2 (BMPR2) gene mutations are a major risk factor for heritable pulmonary arterial hypertension (HPAH), an autosomal dominant fatal disease. We have previously shown that BMPR2 transcripts that contain premature termination codon (PTC) mutations are rapidly and nearly completely degraded through nonsense mediated decay (NMD). Here we report a unique PTC mutation (W13X) that did not behave in the predicted manner. We found that patient-derived cultured lymphocytes (CLs) contained readily detectable levels of the PTC-containing transcript. Further analysis suggested that this transcript escaped NMD by translational re-initiation at a downstream Kozak sequence, resulting in the omission of 173 amino acids. Treatment of CLs containing the PTC with an aminoglycoside decreased the truncated protein levels, with a reciprocal increase in full-length BMPR2 protein and, importantly, BMPR-II signaling. This is the first demonstration of aminoglycoside-mediated 'repair' of a BMPR2 mutation at the protein level in patient-derived cells and has obvious implications for treatment of HPAH where no disease-specific treatment options are available. Our data also suggest the need for a more thorough characterization of mutations prior to labeling them as haploinsufficient or dominant negative based simply on sequencing data.


Hemostasis after transradial coronary intervention by rotary compression device with sterile gauze is associated with more adverse events: a retrospective study.
We investigated the relationship between using a rotary compression device (RCD) with or without sterile gauze and adverse events in transradial access (TRA) for coronary intervention. In this study involving 933 patients at Yueyang Hospital, we recorded TRA-related adverse events, such as bleeding, forearm hematoma, swollen palms, radial artery occlusion (RAO) and others. Logistic regression was applied to assess the association. Of the 933 patients (66.7% males, average age 67.8 years), 511 used RCD with sterile gauze, whereas 422 used RCD without sterile gauze. The most common adverse events were radial artery hemorrhage (7.4%), hand swelling (4.8%) and RAO (4.6%). Logistic regression analysis revealed that the use of RCD with sterile gauze was associated with a higher prevalence of adverse events [odds ratio (OR), 1.74; 95% confidence interval (CI), 1.22-2.49), even with the adjustment of potential confounders (OR, 1.71; 95% CI, 1.19-2.45). Moreover, patients who used RCD with sterile gauze exhibited an increased risk of radial artery hemorrhage (OR, 1.83; 95% CI, 1.07-3.12), swelling of the hand (OR, 1.96; 95% CI, 1.02-3.75) and RAO (OR, 3.17; 95% CI, 1.49-6.72). The use of RCD with sterile gauze in TRA is associated with a higher incidence of adverse events.


Metabolomic assessment of fermentative capability of soybean starter treated with high pressure.
Meju, a brick of dried fermented soybean naturally inoculated with microorganisms, is a starter used for producing traditional Korean fermented soybean products such as soybean paste (doenjang) and soy sauce (ganjang). In order to reduce aging time during production of soybean paste and soy sauce, high pressure (HP) treatment was applied to the meju starter at 500 MPa of pressure for 10 min at 15 degrees C. Fermentative behaviors of normal and HP-treated meju were assessed and compared through physicochemical and (1)H NMR-based metabolomic analysis. All mejues were incubated for 3 weeks at 30 degrees C. At 1 week of incubation, total bacterial population decreased mainly due to a reduction of water content by spontaneous evaporation during the incubation period. As the incubation time increased, glutamate, proline, betain, choline, and phosphocholine levels increased in both normal and HP-treated mejues, indicating that microorganisms in the mejues synthesize these metabolites to endure intracellular hyperosmotic stress induced by the reduction in water content. Through 3 weeks of incubation, the amino-type nitrogen contents and neutral protease activities in HP-treated meju were significantly higher (p < 0.05) than in normal meju, even though total bacterial content in HP-treated meju was 2 or 3 times lower. Moreover, marked increases in glycerol, acetate, tyrosine, and choline levels were observed in HP-treated meju compared to normal meju. In particular, higher levels of tyrosine in HP-treated meju were consistent with the increased neutral protease activities compared to normal meju, indicating an improvement in enzyme stability with HP treatment. These findings highlight a new or better understanding of the influence of the HP or physical treatments on fermentative products in food processing, such as those associated with soybean paste and soy sauce, regarding metabolic behaviors in fermentative starter induced by HP treatment.


Associations between Obstructive Sleep Apnea, Sleep Duration, and Abnormal Fasting Glucose. The Multi-Ethnic Study of Atherosclerosis.
No data exist as to the role of ethnicity in the associations between obstructive sleep apnea (OSA), sleep duration, and metabolic dysfunction. To examine links between OSA, objectively measured habitual sleep duration, and fasting glucose in U.S. ethnic groups. The Multi-Ethnic Study of Atherosclerosis is a multisite community-based study that conducted polysomnography and wrist actigraphy. In 2,151 subjects (1,839 in fully adjusted models), the apnea-hypopnea index was used to classify OSA as none (0-4.9/h), mild (5-14.9/h), or moderate to severe (≥15/h). Actigraphic sleep duration was classified as short (≤5 h/night), intermediate (>5 and <8 h/night), or long (≥8 h/night). Subjects were classified as having normal fasting glucose (<100 mg/dl and no hypoglycemic medication use) or abnormal fasting glucose (≥100 mg/dl and/or hypoglycemic medication use). The sample was 45.8% male, age 68.5 ± 9.2 (mean ± SD) years, and 27.3% African American, 37.2% white, 11.8% Chinese, and 23.8% Hispanic. The prevalence of abnormal fasting glucose was 40.2%. Relative to subjects without apnea, moderate-to-severe OSA was significantly associated with abnormal fasting glucose in African Americans (odds ratio, 2.14; 95% confidence interval, 1.12-4.08) and white participants (odds ratio, 2.85; 95% confidence interval, 1.20-6.75), but not among Chinese or Hispanic subjects, after adjusting for site, age, sex, waist circumference, and sleep duration (P = 0.06 for ethnicity-by-OSA severity interaction). In contrast, sleep duration was not significantly associated with abnormal fasting glucose after considering the influence of OSA. This large multiethnic study confirmed previous reports of an independent association between OSA and metabolic dysfunction, and suggested that this association may vary by ethnicity.


Three-Dimensional Modeling Analysis of Visceral Arteries and Kidneys during Respiration.
Visceral arteries are commonly involved in endovascular repair of complex abdominal aortic aneurysms (AAAs). To improve repair techniques and reduce long-term complications involving visceral arteries, it is crucial to understand in vivo arterial geometry and the deformations due to visceral organ movement with respiration. This study quantifies deformation of the celiac, superior mesenteric (SMA), and renal arteries during respiration and correlates the deformations with diaphragmatic excursion. Sixteen patients with small AAAs underwent magnetic resonance angiography during inspiratory and expiratory breathholds. From geometric models of the aorta and visceral arteries, vessel length, branch angle, curvature, and positions were computed, along with degree of diaphragmatic excursion as indicated by kidney translation. From inspiration to expiration, the celiac artery exhibited axial shortening of 4.8 ± 6.4% (P < 0.001) and a mean curvature increase of 0.03 ± 0.02 mm(-1), greater than other visceral arteries (P < 0.01). With expiration, the SMA, left and right renal arteries (LRA and RRA) angled upward by -9.8 ± 6.4°, -6.4 ± 6.4°, and -5.2 ± 5.0°, respectively (P < 0.005). All vessels translated superiorly (P < 0.0005) and posteriorly (P < 0.01), and the SMA translated rightward additionally (P < 0.005). The left and right kidneys translated by 22 ± 9 mm and 21 ± 9 mm, mostly superiorly (P < 0.001). Translations of all visceral arteries were moderately correlated to the right kidney (R > 0.50). Correlation of the LRA with the left kidney was greater than that of the RRA with the right kidney. The celiac artery exhibited less branch angle change, and greater axial and curvature deformations than the other visceral arteries, due to the vicinity to the liver and influence of the median arcuate ligament. Correlation between visceral arteries and kidney translations revealed that diaphragmatic excursion affects vessel mobility. Weaker correlation of the RRA to the right kidney indicates mechanical shielding from the inferior vena cava.


Doing housework and having regular daily routine standing out as factors associate with physical function in the older people.
Nationwide data were used to explore factors associated with physical function in order to identify interventions that could improve and maintain physical function in the older people. The physical function was assessed by gait speed (GS). We selected 2,677 male and 2,668 female older adults (aged ≥60) who could perform the GS test as study subjects. GS was measured by having subjects walk across and back a 10-m course. A gait speed less than 20% that of a reference population (<0.7 m/s) was used as the definition of slow gait speed (SGS). Co-morbidity, polypharmacy, medical expenses, need for care, and hospitalization were used to evaluate health status. A stepwise logistic regression model was used to determine factors associated with SGS. SGS was associated with poorer health status, higher medical cost, lower ranking on the Geriatric Depression Scale (GDS) and decreased Mini-mental State Examination (MMSE). Co-morbidity (OR = 1.81, 1.58-2.07), polypharmacy (OR = 1.47, 1.25-1.74), MMSE <24 (OR = 1.85, 1.54-2.22), and GDS ≥ 11 (OR = 1.40, 1.18-1.65) were associated with SGS. In contrast, doing housework (DHW, OR = 0.43, 0.38-0.49), having a regular daily routine (RDR, OR = 0.64, 0.45-0.91), and current alcohol consumption (OR = 0.74, 0.62-0.90) were inversely associated with SGS. DHW plus having RDR could greatly reduce the risk of SGS (OR = 0.29, 0.19-0.43). Poor physical function is associated with poorer health status in Chinese older people. Maintaining a regular daily routine and doing some housework may be important factors that can help older people preserve their physical function.


Evidence for Involvement of IL-9 and IL-22 in Cows' Milk Allergy in Infants.
Although allergic inflammation is characterized by a T helper (Th) 2-dominant immune response, the discovery of a role for new T cell subsets in inflammatory diseases has added an additional layer of complexity to the understanding of the pathogeneses of allergic diseases. We evaluated plasma cytokine profiles in infants with cows' milk allergy (CMA), who were being treated with an elimination diet. In a prospective, randomized and controlled study, infants (aged 8.4 ± 3.9 months) with CMA were treated with an elimination diet for 120 days, which replaced cows' milk with a hydrolysed soy protein formula (n = 26) or a free amino acid formula (n = 20). Blood samples were collected before treatment during active disease (T0) and after 120 days, when symptoms were absent (T1). Plasma cytokine concentrations were measured. Infants with CMA had higher plasma concentrations of interleukin (IL)-4 and IL-13 and lower concentrations of IL-9, IL-17A and interferon-γ, compared with healthy breast-fed infants. At T0, there was a positive correlation between blood eosinophil numbers and plasma concentrations of IL-4, IL-9, IL-17A and IL-22. Treatment with a cows' milk elimination diet resulted in a decrease in plasma IL-4, IL-9, IL-13 and IL-22 and an increase in plasma IL-17A. We conclude that IL-4 and IL-13 are elevated in active CMA. The association of IL-9 and IL-22 with eosinophilia, and the decrease in these two cytokines with cows' milk elimination, suggests that they both play a role in the symptoms observed in CMA and may be important targets for future interventions.


Evaluation of the reporting quality of clinical practice guidelines on pancreatic cancer using the RIGHT checklist.
The International Reporting Items for Practice Guidelines in Health Care (RIGHT) statement is a set of recommendations for the reporting in clinical practice guidelines (CPGs). We aimed to assess the reporting quality of CPGs for pancreatic cancer following the RIGHT checklist. Guidelines for pancreatic cancer were identified by searching electronic databases, guideline databases, and medical society websites. The reporting quality was evaluated by calculating the adherence to the items of the RIGHT checklist and summarizing them over the seven domains and the entire checklist. We also present results stratified by selected characteristics. A total of 22 guidelines were found eligible. Mean overall adherence to the RIGHT items was 60.0%. All guidelines adhered to the RIGHT items 3, 7a, 13a, while no guidelines reported the items 14c or 18b, which are some of the topics dealing with rationale for recommendations and funding source, respectively. Of the seven domains of the RIGHT checklist, "Review and quality assurance" and "Funding and declaration and management of interests" had the lowest reporting rates (25.0% and 43.2%, respectively); the remaining five domains had reporting rates >50%. CPGs that reported funding support, were published in higher-impact journals, and that applied a grading system for the quality of evidence, tended to have higher reporting rates. Our results show that reporting quality of pancreatic cancer CPGs still needs to be improved. The use of the RIGHT statement should be encouraged when developing new guidelines.


Technical ability of force application as a determinant factor of sprint performance.
We transposed the concept of effectiveness of force application used in pedaling mechanics to calculate the ratio of forces (RF) during sprint running and tested the hypothesis that field sprint performance was related to the technical ability to produce high amounts of net positive horizontal force. This ability represents how effectively the total force developed by the lower limbs is applied onto the ground, despite increasing speed during the acceleration phase. Twelve physically active male subjects (including two sprinters) performed 8-s sprints on a recently validated instrumented treadmill, and a 100-m sprint on an athletics track. Mean vertical (FV), net horizontal (FH), and total (FTot) ground reaction forces measured at each step during the acceleration allowed computation of the RF as FH/FTot and an index of force application technique (DRF) as the slope of the RF-speed linear relationship from the start until top speed. Correlations were tested between these mechanical variables and field sprint performance variables measured by radar: mean and top 100-m speeds and 4-s distance. Significant (r > 0.731; P < 0.01) correlations were obtained between DRF and 100-m performance (mean and top speeds; 4-s distance). Further, FH was significantly correlated (P < 0.05) to field sprint performance, but FTot and FV were not. Force application technique is a determinant factor of field 100-m sprint performance, which is not the case for the amount of total force subjects are able to apply onto the ground. It seems that the orientation of the total force applied onto the supporting ground during sprint acceleration is more important to performance than its amount.


Quality of T-Cell Response to SARS-CoV-2 mRNA Vaccine in ART-Treated PLWH.
We investigated specific humoral and T-cell responses in people living with HIV (PLWH) before (T0), after two (T1) and after six months (T2) from the third dose of the BNT162b2 vaccine. Healthy donors (HD) were enrolled. The specific humoral response was present in most PLWH already after the second dose, but the third dose increased both the rate of response and its magnitude. Collectively, no significant differences were found in the percentage of responding T-cells between PLWH and HD. At T0, stratifying PLWH according to CD4 cell count, a lower percentage of responding T-cells in <200 cells/µL subgroup compared to >200 cells/µL one was observed. At T1, this parameter was comparable between the two subgroups, and the same result was found at T2. However, the pattern of co-expression of IFNγ, IL2 and TNFα in PLWH was characterized by a higher expression of TNFα, independently of CD4 cell count, indicating a persistent immunological signature despite successful ART. mRNA vaccination elicited a specific response in most PLWH, although the cellular one seems qualitatively inferior compared to HD. Therefore, an understanding of the T-cell quality dynamic is needed to determine the best vaccination strategy and, in general, the capability of immune response in ART-treated PLWH.


[Counseling in artificial fertilization].
Since 1 October 1990, German legislation requires compulsory counselling of an infertile couple before artificial insemination on the medical and psychosocial aspects, by a doctor not performing the treatment. A prerequisite of successful counselling, which enables the couple to take a decision and helps coping with the psychosocial stress of sterility and its consequences, is, in addition to the intrinsic motivation of the couple, also the empathy and the discussion with the doctor. Subject for counselling should be the burdens of therapy, effects of the sterility upon sexuality, partnership, and communication, motivations for wanting a child and the functions of the child for both partners individually and as a partnership. As a prophylactic measure against the development of a neurotic desire for a child and a fixation upon sterility therapy, alternative resolving strategies for therapy failure should be discussed and developed.


PI3K isoform inhibition associated with anti Bcr-Abl drugs shows in vitro increased anti-leukemic activity in Philadelphia chromosome-positive B-acute lymphoblastic leukemia cell lines.
B-acute lymphoblastic leukemia (B-ALL) is a malignant disorder characterized by the abnormal proliferation of B-cell progenitors. Philadelphia chromosome-positive (Ph+) B-ALL is a subtype that expresses the Bcr-Abl fusion protein which represents a negative prognostic factor. Constitutive activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) network is a common feature of B-ALL, influencing cell growth and survival. In the present study, we aimed to investigate the efficacy of PI3K isoform inhibition in B-ALL cell lines harboring the Bcr-Abl fusion protein.We studied the effects of anti Bcr-Abl drugs Imatinib, Nilotinib and GZD824 associated with PI3K isoform inhibitors. We used a panel of six compounds which specifically target PI3K isoforms including the pan-PI3K inhibitor ZSTK474, p110α BYL719 inhibitor and the dual p110γ/p110δ inhibitor IPI145. The effects of single drugs and of several drug combinations were analyzed to assess cytotoxicity by MTS assays, apoptosis and autophagy by flow cytometry and Western blot, as well as the phosphorylation status of the pathway.ZSTK474, BYL719 and IPI145 administered in combination with imatinib, nilotinib and GZD824 for 48 h, decreased cell viability, induced apoptosis and autophagy in a marked synergistic manner.These findings suggest that selected PI3K isoform inhibitors used in combination with anti Bcr-Abl drugs may be an attractive novel therapeutic intervention in Ph+ B-ALL.


Vibrational Spectra of Highly Anharmonic Water Clusters: Molecular Dynamics and Harmonic Analysis Revisited with Constrained Nuclear-Electronic Orbital Methods.
Vibrational spectroscopy is widely used to gain insights into structural and dynamic properties of chemical, biological, and materials systems. Thus, an efficient and accurate method to simulate vibrational spectra is desired. In this paper, we justify and employ a microcanonical molecular simulation scheme to calculate the vibrational spectra of three challenging water clusters: the neutral water dimer (H4O2), the protonated water trimer (H7O3+), and the protonated water tetramer (H9O4+). We find that with the accurate description of quantum nuclear delocalization effects through the constrained nuclear-electronic orbital framework, including vibrational mode coupling effects through molecular dynamics simulations can additionally improve the vibrational spectrum calculations. In contrast, without the quantum nuclear delocalization picture, conventional ab initio molecular dynamics may even lead to less accurate results than harmonic analysis.


The pattern recognition receptor Dectin-1: from fungi to mycobacteria.
The ability of the innate immune system to quickly recognize and respond to an invading pathogen is essential for controlling the infection. For this purpose, cells of the immune system express receptors which recognize evolutionarily conserved structures expressed by various pathogens but absent from host cells. In this review we focus on the non-classical C-type lectin receptors including Dectin-1 whose role has been extensively characterized in the recognition and response to fungal pathogens. Dectin-1 is a type II transmembrane protein which binds beta-1,3 and beta-1,6 glucans. It is expressed on most cells of the innate immune system and has been implicated in phagocytosis as well as killing of fungi by macrophages, neutrophils and dendritic cells. The Dectin-1 cytoplasmic tail contains an immunoreceptor tyrosine based activation motif (ITAM) that signals in part through the spleen tyrosine kinase and in collaboration with Toll-like receptors. Although the main research focus has been on Dectin-1's role as a fungal and yeast pathogen recognition receptor, more recent studies suggest that Dectin-1 may have a broader function in pathogen recognition including a role in directing a macrophage response to mycobacterial infections.


Prevalence of Obesity, No Leisure-Time Physical Activity, and Short Sleep Duration Among Occupational Groups in 29 States.
The aim of this study was to examine prevalence of obesity (body mass index of 30 or higher), no leisure-time physical activity in the past 30 days (no LTPA), and short sleep duration (averaging less than 7 hours of sleep per 24-hour period) among 22 occupational groups. We analyzed 2013 and 2014 Behavioral Risk Factor Surveillance System (BRFSS) data from 29 states, controlling for sex, age, race/ethnicity, and education. By occupation, prevalence ranged from 16.1% to 35.8% for obesity, 11.3% to 28.7% for no LTPA, and 31.4% to 42.9% for short sleep. Only Transportation & Material Moving ranked among the top five occupations for all three risk factors. Obesity and no LTPA varied significantly by sex for several occupations. Prevalence of obesity, no LTPA, and short sleep varied by occupation and affected more than one in five U.S. workers.


Nontransmural laser treatment incompletely denervates canine myocardium.
Clinical experience with transmyocardial laser revascularization (TMLR) has reproducibly demonstrated an improvement in angina class. Denervation has been implicated as a mechanism whereby this clinical effect may be achieved. Because endovascular techniques for TMLR are currently under development, we investigated the impact of nontransmural endoventricular laser treatment on cardiac nerves in a canine model. Five mongrel dogs underwent creation of nontransmural endoventricular channels in the anterior left ventricle with a Holmium:YAG laser. Cardiac afferent nerve function was assessed in control and treatment regions by the epicardial application of bradykinin, a potent algesic, at initial thoracotomy before laser treatment, and at repeat thoracotomy 2 weeks later. The resulting central nervous system-mediated decrease in systemic mean arterial pressure seen in all animals at baseline was reduced by 90% at 2 weeks in the laser-treated territory but was preserved in controls. Immunoblot analysis of tissue samples taken from laser-treated regions demonstrated a 66% reduction in tyrosine hydroxylase, a sympathetic nerve-specific enzyme, as assessed by densitometry. Enzyme content was unchanged in control regions. These data suggest that nontransmural endoventricular laser treatment only partially denervates the heart. This may have implications for the clinical efficacy of the endovascular approach in the relief of angina pectoris.


Intriguing transport dynamics of ethylammonium nitrate-acetonitrile binary mixtures arising from nano-inhomogeneity.
Binary mixtures of ethylammonium nitrate and acetonitrile show interesting properties that originate from the structural and dynamical nano-heterogeneity present in ionic liquids. These effects are most pronounced when the ionic liquid is the minority compound. In this study the transport properties of such mixtures are studied, including viscosity, self-diffusion and conductivity. The results strongly support the presence of structural inhomogeneity and show an interesting composition-dependent behaviour in the mixtures.


[Clinical-EMG characteristic of myasthenia gravis associated with thyroid pathology].
On the basis of clinical-EMG analyses, the researches have revealed the criteria characteristic of myasthenia associated with thyroid pathologies. The myasthenia process associated with thyroid pathology is mainly characterized as of generalized average intensity. Alongside with mup-s with reduced duration and amplitude characteristic of myasthenia, EMG exposes potentials with comparatively long duration and high amplitude (mainly when the process is associated with thyreotoxycosis). It should also be noted that spontaneous activity is intensively expressed. Considering all the above mentioned, we can conclude that these changes show disfunctioning of mup-s rather than the character of synapses caused by the process associated with the myasthenia. The results of the research will facilitate the timely revelation of the process associated with myasthenia and will lead to the unfailing adequate pathogenetic treatment.


Growth of a non-methanotroph on natural gas: ignoring the obvious to focus on the obscure.
Methanotrophs are well known for their ability to grow on methane in natural gas environments; however, these environments also contain low concentrations of longer-chain-length gaseous alkanes. This mixture of alkanes poses a problem for organisms that might otherwise grow on alkanes ≥ C2 because methane could inhibit oxidation of growth substrates and lead to an accumulation of toxic C1 metabolites. Here, we have characterized the growth of a C2 -C9 alkane-utilizing bacterium, Thauera butanivorans, in conditions containing high concentrations of methane and small amounts (< 3% of total alkane) of C2 -C4 . During such growth, methanol accumulates transiently before being consumed in an O2 -dependent process that leads to the formation of a proton gradient and subsequent ATP generation. In contrast, formaldehyde-dependent O2 consumption is insensitive to uncouplers and does not lead to significant ATP production. This efficient C1 oxidation process that regains much of the energy loss inflicted by oxidizing methane, coupled with an alkane monooxygenase effective at limiting methane oxidation, allows T. butanivorans to grow uninhibited in natural gas environments. Although longer-chain-length gaseous alkane-utilizing organisms have been previously identified to grow in natural gas seepages, the data presented here represent the first detailed characterization of the physiological effects associated with inadvertent methane oxidation by a non-methanotroph, and suggest the presence of a well-evolved series of biochemical processes that allow them to grow in natural gas deposits without the need for developing the unique metabolic machinery characteristic of methanotrophs.


Second harmonic generation of shear waves in crystals.
Nonlinear self-interaction of shear waves in electro-elastic crystals is investigated based on the rotationally invariant state function. Theoretical analyses are conducted for cubic, hexagonal, and trigonal crystals. The calculations show that nonlinear self-interaction of shear waves has some characteristics distinctly different from that of longitudinal waves. First, the process of self-interaction to generate its own second harmonic wave is permitted only in some special wave propagation directions for a shear wave. Second, the geometrical nonlinearity originated from finite strain does not contribute to the second harmonic generation (SHG) of shear waves. Therefore, unlike the case of longitudinal wave, the second-order elastic constants do not involve in the nonlinear parameter of the second harmonic generation of shear waves. Third, unlike the nonlinearity parameter of the longitudinal waves, the nonlinear parameter of the shear wave exhibits strong anisotropy, which is directly related to the symmetry of the crystal. In the calculations, the electromechanical coupling nonlinearity is considered for the 6 mm and 3 m symmetry crystals. Complement to the SHG of longitudinal waves already in use, the SHG of shear waves provides more measurements for the determination of third-order elastic constants of solids. The method is applied to a Z-cut lithium niobate (LiNbO3) crystal, and its third-order elastic constant c444 is determined.


Development and loss of early projections in a sexually dimorphic rat spinal nucleus.
The spinal nucleus of the bulbocavernosus (SNB) contains 3-4 times more motoneurons in adult male rats compared to females. This large dimorphism in motoneuron number is produced perinatally by an androgen-regulated cell death. To determine if the early projections of the SNB to its target musculature may be involved in the creation of this sexual dimorphism, and how these projections might interact with androgens, HRP tracing techniques were used to retrogradely label SNB motoneurons during prenatal and postnatal development in males, females, and masculinized females. HRP labeling revealed that the prenatal formation of early projections of the SNB in males and females is comparable. SNB motoneuron number increases through the day before birth in all groups, and during this increase, labeled cells can be seen outside the SNB, which we hypothesize are in the process of migrating into the SNB from the lateral motoneuron column. Postnatally, SNB motoneuron number declines, especially in females, and by postnatal day 10 the sexual dimorphism in cell number and projections has been established. These results indicate that although masculine androgen levels are critical in determining SNB motoneuron survival, they are not necessary for initial axon outgrowth of SNB motoneurons. However, androgens may be involved in the regulation of SNB motoneuron migration and the stabilization of the peripheral projections of the SNB. Both male and female SNB motoneuron axons are present at their target muscles during the time in which sex differences in motoneuron number develop, suggesting that the interaction of SNB motoneurons with their targets could be involved in the dimorphic regulation of cell survival.


IRGM is a common target of RNA viruses that subvert the autophagy network.
Autophagy is a conserved degradative pathway used as a host defense mechanism against intracellular pathogens. However, several viruses can evade or subvert autophagy to insure their own replication. Nevertheless, the molecular details of viral interaction with autophagy remain largely unknown. We have determined the ability of 83 proteins of several families of RNA viruses (Paramyxoviridae, Flaviviridae, Orthomyxoviridae, Retroviridae and Togaviridae), to interact with 44 human autophagy-associated proteins using yeast two-hybrid and bioinformatic analysis. We found that the autophagy network is highly targeted by RNA viruses. Although central to autophagy, targeted proteins have also a high number of connections with proteins of other cellular functions. Interestingly, immunity-associated GTPase family M (IRGM), the most targeted protein, was found to interact with the autophagy-associated proteins ATG5, ATG10, MAP1CL3C and SH3GLB1. Strikingly, reduction of IRGM expression using small interfering RNA impairs both Measles virus (MeV), Hepatitis C virus (HCV) and human immunodeficiency virus-1 (HIV-1)-induced autophagy and viral particle production. Moreover we found that the expression of IRGM-interacting MeV-C, HCV-NS3 or HIV-NEF proteins per se is sufficient to induce autophagy, through an IRGM dependent pathway. Our work reveals an unexpected role of IRGM in virus-induced autophagy and suggests that several different families of RNA viruses may use common strategies to manipulate autophagy to improve viral infectivity.