Results of a prospective strategy to diagnose, localize, and resect insulinomas.
Since 1982, 25 consecutive patients with benign sporadic (non-multiple endocrine neoplasia type I) insulinomas have been studied. Most were referred because either the tumor was not identified at the referring institution or the diagnosis was unclear. Each patient suffered severe neuroglycopenic symptoms for a median of 24 months before diagnosis of insulinoma, and 32% had hypoglycemic seizures. Eighteen patients (72%) had a confirmed weight gain. Each patient underwent a supervised fast until 72 hours or the onset of significant neuroglycopenic symptoms (median duration 16 hours), with serum levels of glucose (median 35 mg/dl; range 24 to 46 mg/dl), insulin (median 21 microU/ml; range 11 to 230 microU/ml), C-peptide (median 2.5 ng/ml; range 1.0 to 7.2 ng/ml), and proinsulin fraction (median 55%; range 14% to 86%) measured at the termination of the fast. Preoperative imaging with ultrasonography, computed tomography, magnetic resonance, and angiography visualized tumor in a minority of patients (26%, 17%, 25%, and 35%, respectively); in 48% of patients one or more imaging study results was positive. Selective portal venous sampling for insulin was the most informative localizing test (77% positive; no false-positive results). Tumor was resected for cure in 24 of 25 patients. Intraoperative ultrasonography identified nonpalpable tumor in seven patients and was crucial to the achievement of this high rate of surgical cure. We conclude that the diagnosis of insulinoma can be made by the results of a supervised fast, portal venous sampling is the most sensitive preoperative test for localizing insulinomas, and intraoperative ultrasonography is essential for intraoperative detection of insulinomas.


ERG voltage-gated K+ channels regulate excitability and discharge dynamics of the medial vestibular nucleus neurones.
The discharge properties of the medial vestibular nucleus neurones (MVNn) critically depend on the activity of several ion channel types. In this study we show, immunohistochemically, that the voltage-gated K(+) channels ERG1A, ERG1B, ERG2 and ERG3 are highly expressed within the vestibular nuclei of P10 and P60 mice. The role played by these channels in the spike-generating mechanisms of the MVNn and in temporal information processing was investigated electrophysiologically from mouse brain slices, in vitro, by analysing the spontaneous discharge and the response to square-, ramp- and sinusoid-like intracellular DC current injections in extracellular and whole-cell patch-clamp studies. We show that more than half of the recorded MVNn were responsive to ERG channel block (WAY-123,398, E4031), displaying an increase in spontaneous activity and discharge irregularity. The response to step and ramp current injection was also modified by ERG block showing a reduction of first spike latency, enhancement of discharge rate and reduction of the slow spike-frequency adaptation process. ERG channels influence the interspike slope without affecting the spike shape. Moreover, in response to sinusoid-like current, ERG channel block caused frequency-dependent gain enhancement and phase-lead shift. Taken together, the data demonstrate that ERG channels control the excitability of MVNn, their discharge regularity and probably their resonance properties.


[Myositis: current data on the classification, diagnosis and treatment].
The classification of idiopathic inflammatory myopathies has been refined with the characterization of new antibodies and their clinicopathological associations. The most widely used classification (Troyanov) defines pure polymyositis (PM) and dermatomyositis (DM), myositis overlap (presence of extra-muscular-extra-cutaneous manifestations or auto-antibodies), myositis associated to cancers and sporadic inclusion body myositis IBM). Overlap myositis are generally more severe and chronic than pure forms, and almost always require immunosuppressants. IBM remains difficult to treat, but immunosuppressants or immunoglobulins may be proposed, especially at the beginning of evolution.


Comparative digestibility of energy and nutrients in diets fed to sows and growing pigs.
The objective of this research was to compare values for digestible energy (DE) and metabolisable energy (ME) and apparent total tract digestibility (ATTD) of nutrients in 11 diets fed to both growing pigs and gestating sows. Three diets were based on corn, wheat or sorghum and eight diets were based on a combination of corn and soybean meal, canola meal, conventional distillers' dried grains with solubles, low-fat distillers' dried grains with solubles, corn germ meal, corn bran, wheat middlings or soybean hulls. A total of 88 gestating sows (252 ± 24.2 kg BW; parity two to six) and 88 growing barrows (40 ± 4.7 kg BW) were used and randomly allotted to the 11 diets with eight replicate sows or pigs per diet. Faecal and urine samples were collected for 4 d following a 19 d adaptation period. The DE, ME and ATTD of gross energy (GE), acid detergent fibre (ADF), neutral detergent fibre (NDF) and crude protein (CP) in the 11 diets were calculated. Gestating sows had greater (p < 0.05) ATTD of GE and CP and DE values for all diets compared with growing pigs. Gestating sows also had greater (p < 0.05) ME values than growing pigs for the three grain diets and the diets containing wheat middlings and soybean hulls. No differences were observed in ATTD of ADF and NDF between gestating sows and growing pigs for any of the diets, except that gestating sows had greater (p < 0.05) ATTD of NDF than growing pigs when they were fed the four protein diets. The ATTD of GE and CP and DE values in gestating sows may be predicted by using equations generated from the values of ATTD of GE and CP and DE values obtained in growing pigs. Results of this research indicate that ATTD values of CP and GE obtained in gestating sows are greater than the values obtained in growing pigs, but values for ATTD of ADF obtained in growing pigs are not different from values in gestating sows.


[Immunoepidemiological characteristics of the duration of a meningococcal carrier state].
The duration of meningococcal carriership in children and adults in the foci of infection and outside such foci and the immunological characteristics relating to group-specific meningococcal antigens A, C, X, Y and Z at different periods after the detection of the infective agent in the nasopharynx have been studied. Carrier state has been shown to last, on the average, 11 days. The duration of the release of meningococci from the nasopharynx has proved to be influenced by the epidemic situation in a given group. Differences in the time course of the immunological reorganization of the body in response to antigenic challenge in prolonged and short-term carrier state have been detected. These data suggest that rapid immune response to meningococcal antigens in the process of short-term carrier state is probably one of the factors preventing the prolonged colonization of the nasopharynx by the infective agent.


Adaptation to novel accents by toddlers.
Word recognition is a balancing act: listeners must be sensitive to phonetic detail to avoid confusing similar words, yet, at the same time, be flexible enough to adapt to phonetically variable pronunciations, such as those produced by speakers of different dialects or by non-native speakers. Recent work has demonstrated that young toddlers are sensitive to phonetic detail during word recognition; pronunciations that deviate from the typical phonological form lead to a disruption of processing. However, it is not known whether young word learners show the flexibility that is characteristic of adult word recognition. The present study explores whether toddlers can adapt to artificial accents in which there is a vowel category shift with respect to the native language. 18-20-month-olds heard mispronunciations of familiar words (e.g., vowels were shifted from [a] to [æ]: "dog" pronounced as "dag"). In test, toddlers were tolerant of mispronunciations if they had recently been exposed to the same vowel shift, but not if they had been exposed to standard pronunciations or other vowel shifts. The effects extended beyond particular items heard in exposure to words sharing the same vowels. These results indicate that, like adults, toddlers show flexibility in their interpretation of phonological detail. Moreover, they suggest that effects of top-down knowledge on the reinterpretation of phonological detail generalize across the phono-lexical system.


GABAergic signaling to astrocytes in the prefrontal cortex sustains goal-directed behaviors.
GABA interneurons play a critical role in higher brain functions. Astrocytic glial cells interact with synapses throughout the whole brain and are recognized as regulatory elements of excitatory synaptic transmission. However, it is largely unknown how GABAergic interneurons and astrocytes interact and contribute to stable performance of complex behaviors. Here, we found that genetic ablation of GABAB receptors in medial prefrontal cortex astrocytes altered low-gamma oscillations and firing properties of cortical neurons, which affected goal-directed behaviors. Remarkably, working memory deficits were restored by optogenetic stimulation of astrocytes with melanopsin. Furthermore, melanopsin-activated astrocytes in wild-type mice enhanced the firing rate of cortical neurons and gamma oscillations, as well as improved cognition. Therefore, our work identifies astrocytes as a hub for controlling inhibition in cortical circuits, providing a novel pathway for the behaviorally relevant midrange time-scale regulation of cortical information processing and consistent goal-directed behaviors.


Galectin-3 is a candidate biomarker for amyotrophic lateral sclerosis: discovery by a proteomics approach.
The discovery of biomarkers for neurodegenerative diseases will have a major impact on the efficiency of therapeutic clinical trials and may be important for understanding basic pathogenic mechanisms. We have approached the discovery of protein biomarkers for amyotrophic lateral sclerosis (ALS) by profiling affected tissues in a relevant animal model and then validating the findings in human tissues. Ventral roots from SOD1(G93A) "ALS" mice were analyzed by label-free quantitative mass spectrometry, and the resulting data were compared with data for matched samples from nontransgenic littermates and transgenic mice carrying wild-type human SOD1 (SOD1(WT)). Of 1299 proteins, statistical inference of the data in the three groups identified 14 proteins that were dramatically altered in the ALS mice compared with the two control groups. The protein galectin-3 emerged as a lead biomarker candidate on the basis of its differential expression as assessed by immunoblot and immunocytochemistry in SOD1(G93A) mice as compared to controls and because it is a secreted protein that could potentially be measured in human biofluids. Spinal cord tissue from ALS patients also exhibited increased levels of galectin-3 when compared to controls. Further measurement of galectin-3 in cerebrospinal fluid samples showed that ALS patients had approximately twice as much galectin-3 as normal and disease controls. These results provide the proof of principle that biomarker identification in relevant and well-controlled animal models can be translated to human disease. The challenge is to validate our biomarker candidate proteins as true biomarkers for ALS that will be useful for diagnosis and/or monitoring disease activity in future clinical trials.


[Integral photodynamic therapy of multifocal bladder cancer. Initial clinical experiences].
Experimental results and initial clinical experience indicates that even carcinoma in situ can be destroyed by integral photodynamic therapy of multifocal bladder carcinoma following tumor-selective photosensitization with a hematoporphyrin derivative. The employment of a light-scattering medium in combination with a special bladder catheter and photodynamically effective red laser light (630 nm) guarantees homogeneous irradiation of the bladder, which thus reaches all tumor foci. The complications observed were transitional dysurias and, in a particular case, regressive shrinkage of the bladder.


Incidence of new atherothrombotic brain infarction in older persons with prior myocardial infarction and serum low-density lipoprotein cholesterol >or=125 mg/dl treated with statins versus no lipid-lowering drug.
We report the incidence of new atherothrombotic brain infarction (ABI) in older men and women with prior myocardial infarction and a serum low-density lipoprotein (LDL) cholesterol of >or=125 mg/dl treated with statins and with no lipid-lowering drug. The incidence of new ABI was investigated in an observational prospective study of 1410 men and women, mean age 81 +/- 9 years, with prior myocardial infarction and a serum LDL cholesterol of >or=125 mg/dl treated with statins (679 persons or 48%) and with no lipid-lowering drug (731 persons or 52%). Follow-up was 36 +/- 21 months. At follow-up, the stepwise Cox regression model showed that significant independent predictors of new ABI were age (risk ratio = 1.04 for a 1-year increase in age), cigarette smoking (risk ratio = 3.5), hypertension (risk ratio = 3.1), diabetes mellitus (risk ratio = 2.3), initial serum LDL cholesterol (risk ratio = 1.01 for each 1 mg/dl increase), initial serum high-density lipoprotein cholesterol (risk ratio = 0.97 for each 1 mg/dl increase), prior stroke (risk ratio = 2.5), and use of statins (risk ratio = 0.40). The Cochran-Armitage test showed a trend in the reduction of new ABI in persons treated with statins as the level of serum LDL cholesterol decreased ( p <.0001). Use of statins caused a 60%, significant, independent reduction in new ABI in older men and women with prior myocardial infarction and a serum LDL cholesterol of >or=125 mg/dl.


Trichinosis in sows slaughtered at a Kentucky abattoir.
From July 26 to 30, 1976, diaphragmatic pillar samples were collected from 513 sows slaughtered at a federally inspected abattoir in Kentucky. The sows represented 5 marketing areas: Rushville, In (186); Cincinnati, Oh )158); Indianapolis, In (70); Georgetown, Ky (54); and Columbus, Oh (45). Using the pooled-sample digestion technique, 3 dead trichina larvae were identified in 1 sample pooled from 15 diaphragms (10 g each). An attempt to identify the individually infected sow(s) was unsuccessful, but the pooled sample originated from animals marketed at Rushville, In. The prevalence (1 of 513) was 0.194%.


A deep learning model for predicting risks of crop pests and diseases from sequential environmental data.
Crop pests reduce productivity, so managing them through early detection and prevention is essential. Data from various modalities are being used to predict crop diseases by applying machine learning methodology. In particular, because growth environment data is relatively easy to obtain, many attempts are made to predict pests and diseases using it. In this paper, we propose a model that predicts diseases through previous growth environment information of crops, including air temperature, relative humidity, dew point, and CO2 concentration, using deep learning techniques. Using large-scale public data on crops of strawberry, pepper, grape, tomato, and paprika, we showed the model can predict the risk score of crop pests and diseases. It showed high predictive performance with an average AUROC of 0.917, and based on the predicted results, it can help prevent pests or post-processing. This environmental data-based crop disease prediction model and learning framework are expected to be universally applicable to various facilities and crops for disease/pest prevention.


Identification of the gene lon (capR) product as a 94-kilodalton polypeptide by cloning and deletion analysis.
A mutation in the lon (capR) gene of Escherichia coli K-12 effects several phenotypic alterations in the mutant cell, such as overproduction of capsular polysaccharide and sensitivity to ultraviolet or ionizing radiation. A previously cloned 9.2-megadalton (Md) EcoRI fragment contained the capR+ gene and specified two polypeptides, 94 kilodaltons (K) and 67K. To provide evidence that the 94K polypeptide is the capR+ gene product, we constructed a capR+ plasmid pJMC40, having a 2.0-Md EcoRI-PstI fragment which codes only for the 94K polypeptide. Plasmids pJMC22 and pJMC30, having deletions of 0.7 and 0.8 Md, respectively, from one end of the 2.0-Md fragment, were also constructed. Each codes for a shortened stable polypeptide (from the 94K). Neither plasmid can confer the capR+ phenotype to capR mutants, confirming that the unaltered 94K polypeptide is the capR+ gene product. Plasmids pJMC51 and pJMC52 each have a deletion of 0.7 Md from the other end of the 2.0-Md fragment, differing only in the orientation of the remaining 1.3-Md fragment with respect to the cloning vehicle. They are nonfunctional with respect to capR+ and do not code for a common polypeptide from the 1.3-Md fragment. These data indicate that the fragments in pJMC22 and pJMC30, which both code for shortened 94K polypeptides, contain the promoter-operator region of the capR gene. The deletion plasmids were also used to map chromosomal capR mutations.


Houseflies: not simple mechanical vectors of enterohemorrhagic Escherichia coli O157:H7.
An epidemic of enterohemorrhagic colitis caused by Escherichia coli O157:H7 (EHEC-O157) occurred in a nursery school in a rural area of Japan in September 1996. The EHEC-O157 were isolated both from patients and houseflies collected at the school. The flies were suspected to be mechanical vectors of the pathogen. Feeding experiments of EHEC-O157 to houseflies showed that the ingested bacteria were harbored in the intestine of flies and continued to be excreted at least for 3 days after feeding. Scanning electron microscopy showed that a large number of EHEC-O157 adhered to the surface of the housefly mouthparts and actively proliferated in the minute spaces of the labellum. Food masses containing EHEC-O157 in the fly intestine were completely surrounded by a peritrophic membrane during digestion and discharged rapidly. The persistence of bacteria in the intestine and feces is mainly a result of proliferation in the mouthparts and accumulation in the crop. Our results strongly suggest that houseflies are not simple mechanical vectors of EHEC. The epidemiologic potential of houseflies to disseminate EHEC-O157 may be greater than initially suspected.


The potential pitfalls of studying adult sex ratios at aggregate levels in humans.
Human adult sex ratios have been studied extensively across the biological and social sciences. While several studies have examined adult sex ratio effects in a multilevel perspective, many studies have focused on effects at an aggregated level only. In this paper, we review some key issues relating to such analyses. We address not only nation-level analyses, but also aggregation at lower levels, to investigate whether these issues extend to lower levels of aggregation. We illustrate these issues with novel databases covering a broad range of variables. Specifically, we discuss distributional issues with aggregated measures of adult sex ratio, significance testing, and statistical non-independence when using aggregate data. Firstly, we show that there are severe distributional issues with national adult sex ratio, such as extreme cases. Secondly, we demonstrate that many 'meaningless' variables are significantly correlated with adult sex ratio (e.g. the max. elevation level correlates with sex ratio at US state level). Finally, we re-examine associations between adult sex ratios and teenage fertility and find no robust evidence for an association at the aggregate level. Our review highlights the potential issues of using aggregate data on adult sex ratios to test hypotheses from an evolutionary perspective in humans.This article is part of the themed issue 'Adult sex ratios and reproductive decisions: a critical re-examination of sex differences in human and animal societies'.


Substrate interaction dynamics and oxygen control in the active site of thymidylate synthase ThyX.
Thymidylate synthase ThyX, required for DNA synthesis in many pathogenic bacteria, is considered a promising antimicrobial target. It binds FAD and three substrates, producing dTMP (2'-deoxythymidine-5'-monophosphate) from dUMP (2'-deoxyuridine-5'-monophosphate). However, ThyX proteins also act as NADPH oxidase by reacting directly with O2. In the present study we investigated the dynamic interplay between the substrates and their role in competing with this wasteful and potentially harmful oxidase reaction in catalytically efficient ThyX from Paramecium bursaria Chlorella virus-1. dUMP binding accelerates the O2-insensitive half-reaction between NADPH and FAD by over four orders of magnitude to ~30 s-1. Thus, although dUMP does not have a direct role in FAD reduction, any turnover with molecular O2 requires its presence. Inversely, NADPH accommodation accelerates dUMP binding ~3-fold and apparently precedes dUMP binding under physiological conditions. In the oxidative half-reaction, excess CH2H4folate (N5,N10-methylene-5,6,7,8-tetrahydrofolate) was found to re-oxidize FADH2 within 1 ms, thus very efficiently competing with FADH2 oxidation by O2 (1.5 s-1 under aerobic conditions). The resulting reaction scheme points out how the interplay between the fast reactions with the native substrates, although not rate-limiting for overall catalysis, avoids NADPH oxidase activity in aerobic micro-organisms, including many pathogens. These observations also explain why ThyX proteins are also present in aerobic micro-organisms.


Genetic fine structure analysis of Helicobacter pylori isolates before and after treatment.
Eradication of H. pylori infection cures peptic ulcer disease and conversely, relapse is associated with reappearance of H. pylori infection. However, it is not clear whether the recurrence of ulcers following H. pylori eradication is due to recrudescence (identical strain) of the previous infection or as a result of exogenous reinfection (different strain) by another strain. The aim of the present study was to analyze the FAFLP patterns of pre and post treatment H. pylori samples to check if the recurrence was due to recrudescence or reinfection. 24 of 30 duodenal ulcer (DU) subjects screened for H. pylori infection were positive for H. pylori infection. The treatment regime included pantoprazole, ciprofloxacin and amoxicillin. The patients were called for a repeat endoscopy after one month and screened for H. pylori infection. FAFLP analysis and PCR for the cagA and vacA gene was performed for the pre and post treatment samples. Of the 24 positive H. pylori patients, only 6 were negative after treatment and the remaining 18 were positive for H. pylori infection. The analysis of the pre and post treatment samples of the 18 patients showed that the FAFLP profiles of the initial and follow-up pools were similar to one another. It can be concluded that in the present series of patients, reinfection was due to recrudescence of infection due to incomplete eradication. The study also suggests that DNA fingerprinting by FAFLP provides discriminatory and complementary data for identifying strains of H. pylori while monitoring therapy.


Immune Characteristics Analysis and Transcriptional Regulation Prediction Based on Gene Signatures of Chronic Obstructive Pulmonary Disease.
The variation in inflammation in chronic obstructive pulmonary disease (COPD) between individuals is genetically determined. This study aimed to identify gene signatures of COPD through bioinformatics analysis based on multiple gene sets and explore their immune characteristics and transcriptional regulation mechanisms. Data from four microarrays were downloaded from the Gene Expression Omnibus database to screen differentially expressed genes (DEGs) between COPD patients and controls. Weighted gene co-expression network analysis was applied to identify trait-related modules and then select key module-related DEGs. The optimized gene set of signatures was obtained using the least absolute shrinkage and selection operator (LASSO) regression analysis. The CIBERSORT algorithm and Pearson correlation test were used to analyze the relationship between gene signatures and immune cells. Finally, public databases were used to predict the transcription factors (TFs) and upstream miRNAs. A total of 127 DEGs in COPD were identified from the combined dataset. By considering the intersection of DEGs and genes in two trait-related modules, 83 key module-related DEGs were identified, which were mainly enriched in interleukin-related pathways. Seven-gene signatures, including MTHFD2, KANK3, GFPT2, PHLDA1, HS3ST2, FGG, and RPS4Y1, were further selected using the LASSO algorithm. These gene signatures showed the predictive potential for COPD risks and were significantly correlated with 18 types of immune cells. Finally, nine miRNAs and three TFs were predicted to target MTHFD2, GFPT2, PHLDA1, and FGG. We proposed the seven-gene-signature to predict COPD risk and explored its potential immune characteristics and regulatory mechanisms.


Systematic screening of Drosophila deficiency mutations for embryonic phenotypes and orphan receptor ligands.
This paper defines a collection of Drosophila deletion mutations (deficiencies) that can be systematically screened for embryonic phenotypes, orphan receptor ligands, and genes affecting protein localization. It reports the results of deficiency screens we have conducted that have revealed new axon guidance phenotypes in the central nervous system and neuromuscular system and permitted a quantitative assessment of the number of potential genes involved in regulating guidance of specific motor axon branches. Deficiency "kits" that cover the genome with a minimum number of lines have been established to facilitate gene mapping. These kits cannot be systematically analyzed for phenotypes, however, since embryos homozygous for many deficiencies in these kits fail to develop due to the loss of key gene products encoded within the deficiency. To create new kits that can be screened for phenotype, we have examined the development of the nervous system in embryos homozygous for more than 700 distinct deficiency mutations. A kit of approximately 400 deficiency lines for which homozygotes have a recognizable nervous system and intact body walls encompasses >80% of the genome. Here we show examples of screens of this kit for orphan receptor ligands and neuronal antigen expression. It can also be used to find genes involved in expression, patterning, and subcellular localization of any protein that can be visualized by antibody staining. A subset kit of 233 deficiency lines, for which homozygotes develop relatively normally to late stage 16, covers approximately 50% of the genome. We have screened it for axon guidance phenotypes, and we present examples of new phenotypes we have identified. The subset kit can be used to screen for phenotypes affecting all embryonic organs. In the future, these deficiency kits will allow Drosophila researchers to rapidly and efficiently execute genome-wide anatomical screens that require examination of individual embryos at high magnification.


Regular, low-dose methadone for reducing breathlessness in people experiencing or at risk of neurotoxic effects from morphine: A single-center case series.
Breathlessness is a common symptom suffered by people living with advanced malignant and non-malignant diseases, one which significantly limits their quality of life. If it emerges at minimal exertion, despite the maximal, guidelines-directed, disease-specific therapies, it should be considered persistent and obliges clinicians to prescribe symptomatic, non-pharmacological, and pharmacological treatment to alleviate it. Opioids are recommended for the symptomatic treatment of persistent breathlessness, with morphine most extensively studied for this indication. It is extensively metabolized in the liver into water-soluble 3- and 6-glucuronides, excreted by the kidneys. In the case of advanced renal failure, the glucuronides accumulate, mainly responsible for toxicity 3-glucuronides. Some people, predominantly those with advanced renal failure, develop neurotoxic effects after chronic morphine, even when prescribed at a very low dose. A single-center case series of consecutive patients experiencing neurotoxic effects after long-term, low-dose morphine or at risk of such effects were transferred to methadone to avoid the accumulation of neurotoxic metabolites. Over the course of 4.5 years, 26 patients have been treated with methadone in the median dose of 3.0 mg/24 h p.o., for persisting breathlessness. Sixteen of them had been treated previously with an opioid (usually morphine) at the median dose of 7.0 mg/24 h (morphine oral daily dose equivalent). They were transferred to methadone, with the median dose of 3.0 mg/24 h orally (methadone oral daily dose equivalent), and the median morphine-to-methadone dose ratio was 2.5:1. All patients experienced a meaningful improvement in breathlessness intensity after methadone, by a median of 5 points (range 1-8) on the 0-10 numerical rating scale (NRS) in the whole group, and by 2 points (range 0-8) in those pretreated with other opioids, mainly morphine. Low-dose methadone can be considered an efficient alternative to morphine for reducing breathlessness in people experiencing neurotoxic effects or at risk of developing them following treatment with morphine.


Exoenzyme S of Pseudomonas aeruginosa is secreted by a type III pathway.
Exoenzyme S is an extracellular ADP-ribosyltransferase of Pseudomonas aeruginosa. Transposon mutagenesis of P. aeruginosa 388 was used to identify genes required for exoenzyme S production. Five Tn5Tc insertion mutants were isolated which exhibited an exoenzyme S-deficient phenotype (388::Tn5Tc 469, 550, 3453, 4885, and 5590). Mapping experiments demonstrated that 388::Tn5Tc 3453, 4885, and 5590 possessed insertions within a 5.0 kb EcoRI fragment that is not contiguous with the exoenzyme S trans-regulatory operon. 388::Tn5Tc 469 and 550 mapped to a region downstream of the trans-regulatory operon which has been previously shown to contain a promoter region that is co-ordinately regulated with exoenzyme S synthesis. Nucleotide sequence analysis of a 7.2 kb region flanking the 388::Tn5Tc 469 and 550 insertions, identified 12 contiguous open reading frames (ORFs). Database searches indicated that the first ORF, ExsD, is unique. The other 11 ORFs demonstrated high homology to the YscB-L proteins of the yersiniae Yop type III export apparatus. RNase-protection analysis of wild-type and mutant strains indicated that exsD and pscB-L form an operon. To determine whether ExoS was exported by a type III mechanism, derivatives consisting of internal deletions or lacking amino- or carboxy-terminal residues were expressed in P. aeruginosa. Deletion analyses indicated that the amino-terminal nine residues are required for ExoS export. Combined data from mutagenesis, regulatory, expression, and sequence analyses provide strong evidence that P. aeruginosa possesses a type III secretion apparatus which is required for the export of exoenzyme S and potentially other co-ordinately regulated proteins.


[Ischaemic heart disease and chronic helicobacter pylori infection--present views].
In connexion with the high prevalence of coronary heart disease its additional reasons are searched, which could lead to the occurrence and development of the disease. Chronic infections have been lately recognized as an additional reason. In the article we presented opinions regarding the influence of long-lasting Helicobacter pylori infection on the development of inflammation process, atheromatous changes, initiation and development of ischaemic heart disease.


Distress tolerance and physiological reactivity to stress predict women's problematic alcohol use.
Research has shown that measures of reactivity to distress-including distress tolerance and physiological reactivity to stress-are dysregulated in women who misuse alcohol. These variables may interact and create a risk profile for young adult women, reflecting patterns of stress reactivity that confer a risk for alcohol misuse. The current study tested this hypothesis by examining the independent and interactive associations of subjective distress tolerance, behavioral distress tolerance, and physiological stress reactivity with women's alcohol misuse. The study was conducted with a sample of 91 college women recruited on a large northeastern university campus. Results showed that subjective levels of distress tolerance and physiological reactivity to stress (skin conductance reactivity, SCR), but not behavioral distress tolerance, were independently associated with alcohol misuse. In addition, subjective distress tolerance moderated the relationship between SCR and negative alcohol-related consequences. Specifically, women with low physiological reactivity (SCR) to a stressful task and greater urge to quickly rid themselves of distress (low subjective distress tolerance) endorsed a significantly greater number of adverse consequences from their alcohol use. These results extend prior findings by showing that, even among a nonclinical sample of women, lower stress reactivity in combination with low subjective distress tolerance is associated with increased risk for various drinking-related negative consequences. (PsycINFO Database Record


Standing balance of children with developmental coordination disorder under altered sensory conditions.
Maintenance of standing balance requires that sensory inputs be organized with the motor system. Current data regarding the influence of sensory inputs on standing balance in children with developmental coordination disorder (DCD) are limited. This study compared the influence of sensory organization and each sensory input on the standing stability between a group of 20 children, 4-6 years old, with DCD and an age- and gender-matched control group of 20 children. Three types of visual inputs (eyes open, eyes closed, or unreliable vision) and two types of somatosensory inputs (fixed or compliant foot support) were varied factorially to yield six sensory conditions. Standing stability was measured with a Kistler force plate for 30s and expressed as the center of pressure sway area. The results showed that the standing stability of the children with DCD was significantly poorer than that of the control children under all sensory conditions, especially when the somatosensory input was unreliable (compliant foot support) compared to when it was reliable (fixed foot support). The effectiveness of an individual sensory system, when it was the dominant source of sensory input, did not significantly differ between the groups. The results suggest that children with DCD experience more difficulty coping with altered sensory inputs, and that such difficulty is more likely due to a deficit in sensory organization rather than compromised effectiveness of individual sensory systems.


A qualitative study of patient motivation to adhere to combination antiretroviral therapy in South Africa.
Taken as prescribed, that is, with high adherence, combination antiretroviral therapy (ART) has changed HIV infection and disease from being a sure predictor of death to a manageable chronic illness. Adherence, however, is difficult to achieve and maintain. The CAPRISA 058 study was conducted between 2007 and 2009 to test the efficacy of individualized motivational counselling to enhance ART adherence in South Africa. As part of the overall trial, a qualitative sub-study was conducted, including 30 individual interviews and four focus group discussions with patients in the first 9 months of ART initiation. Data were inductively analyzed, using thematic analysis, to identify themes central to ART adherence in this context. Four themes emerged that characterize the participants' experiences and high motivation to adhere to ART. Participants in this study were highly motivated to adhere, as they acknowledged that ART was 'life-giving', in the face of a large amount of morbidity and mortality. They were further supported by techniques of routine remembering, and highlighted the importance of good social support and access to supportive healthcare workers, to their continued success in negotiating their treatment. Participants in the current study told us that their adherence motivation is enhanced by free accessible care, approachable and supportive healthcare workers, broad social acceptance of ART, and past first-hand experiences with AIDS-related co-morbidity and mortality. Programs that include specific attention to these aspects of care will likely be successful in the long term.


Ketogenic Diet Therapy in Infants: Efficacy and Tolerability.
This study evaluated tolerability and efficacy of the ketogenic diet in infants less than 12 months of age. Infants less than 12 months of age, commencing the ketogenic diet between September 2007 and July 2016 were identified. Records were reviewed for epilepsy details, diet initiation details, efficacy and tolerability. Twenty-seven infants commenced the ketogenic diet (56% male, median age seven months). Median age at seizure onset was 1.9 months and 92% had daily seizures. An epilepsy syndrome was noted in 19 (West-11, epilepsy in infancy with migrating focal seizures-5, early myoclonic encephalopathy-1, Ohtahara-1, Dravet-1). Infants were on a median of two and had failed a median of one medications for lack of efficacy. All initiated a traditional ketogenic diet at full calories without fasting, and all but one started the diet in hospital. Significant hypoglycemia during initiation was seen in two - both had emesis +/- decreased oral intake. Eighty-eight percent developed urinary ketosis by 48 hours and all were successfully discharged on the diet (median ratio 3:1). Of those continuing dietary therapy, responder rates at one, six and 12 months were 68%, 82% and 91%, with 20%, 29% and 27% achieving seizure freedom. By 12 months, two stopped the diet for serious adverse effects, five discontinued for lack of efficacy, six were lost to follow-up and two died of unrelated causes. The ketogenic diet is an effective and well-tolerated treatment for infants with intractable epilepsy. In-hospital initiation is strongly recommended due to risk of hypoglycemia with emesis or reduced intake.


Diurnal variations in lower leg subcutaneous blood flow rate in patients with chronic venous leg ulcers.
The blood flow rate in subcutaneous adipose tissue was measured on the lower legs of 11 patients with chronic lower-leg venous insufficiency and ulceration and in eight age-matched control subjects for 12-20 h, under ambulatory conditions, using the 133Xe wash-out technique with portable Cadmium telluride (CdTe(Cl)) detectors. In both groups, the change from an upright to a supine position at the beginning of the night period elicited an instantaneous increment in the blood flow rate of 30-40% with a decrease in the central and local postural sympathetic vasoconstrictor activity. After approximately 1 h of sleep, a considerable increase in blood flow rate was seen in both patient and control groups which persisted for nearly 100 min. In the patient group, the mean increase was 137% compared to a mean increase of 68% in the control group (P less than 0.01). The blood flow then returned to the same level as at the beginning of the night and remained stable until the subjects woke in the morning. The differences between this nocturnal hyperaemic phase and the adjacent phases were highly significant in both groups (P less than 0.0001 and P less than 0.005, respectively). The blood flow levels measured on the second day were of the same magnitude as those registered on the first day. The mean values of the wash-out rate constants (k) of the two groups were of the same magnitude in all phases of subcutaneous blood flow level, except in the hyperaemic phase, where it was significantly higher in the patient group (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Generation and Evolution of Neural Cell Types and Circuits: Insights from the Drosophila Visual System.
The Drosophila visual system has become a premier model for probing how neural diversity is generated during development. Recent work has provided deeper insight into the elaborate mechanisms that control the range of types and numbers of neurons produced, which neurons survive, and how they interact. These processes drive visual function and influence behavioral preferences. Other studies are beginning to provide insight into how neuronal diversity evolved in insects by adding new cell types and modifying neural circuits. Some of the most powerful comparisons have been those made to the Drosophila visual system, where a deeper understanding of molecular mechanisms allows for the generation of hypotheses about the evolution of neural anatomy and function. The evolution of new neural types contributes additional complexity to the brain and poses intriguing questions about how new neurons interact with existing circuitry. We explore how such individual changes in a variety of species might play a role over evolutionary timescales. Lessons learned from the fly visual system apply to other neural systems, including the fly central brain, where decisions are made and memories are stored.


Increased H3K9me3 drives dedifferentiated phenotype via KLF6 repression in liposarcoma.
Liposarcoma (LPS) can be divided into 4 different subtypes, of which well-differentiated LPS (WDLPS) and dedifferentiated LPS (DDLPS) are the most common. WDLPS is typically low grade, whereas DDLPS is high grade, aggressive, and carries a worse prognosis. WDLPS and DDLPS frequently co-occur in patients. However, it is not clear whether DDLPS arises independently from WDLPS, or whether epigenomic alterations underly the histopathological differences of these subtypes. Here, we profiled 9 epigenetic marks in tumor samples from 151 patients with LPS and showed elevated trimethylation of histone H3 at Lys9 (H3K9me3) levels in DDLPS tumors. Integrated ChIP-seq and gene expression analyses of patient-derived cell lines revealed that H3K9me3 mediates differential regulation of genes involved in cellular differentiation and migration. Among these, Kruppel-like factor 6 (KLF6) was reduced in DDLPS, with increased H3K9me3 at associated regulatory regions. Pharmacologic inhibition of H3K9me3 with chaetocin decreased DDLPS proliferation and increased expression of the adipogenesis-associated factors PPARγ, CEBPα, and CEBPβ, suggesting that increased H3K9me3 may mediate DDLPS-associated aggressiveness and dedifferentiation properties. KLF6 overexpression partially phenocopied chaetocin treatment in DDLPS cells and induced phenotypic changes that were consistent with adipocytic differentiation, suggesting that the effects of increased H3K9me3 may be mediated through KLF6. In conclusion, we provide evidence of an epigenetic basis for the transition between WDLPS and DDLPS.


Spin the light off: rapid internal conversion into a dark doublet state quenches the fluorescence of an RNA spin label.
The spin label Çm and the fluorophore Çmf are close isosteric relatives: the secondary amine Çmf can be easily oxidized to a nitroxide group to form Çm. Thus, both compounds can serve as EPR and fluorescence labels, respectively, and their high structural similarity allows direct comparison of EPR and fluorescence data, e.g. in the context of investigations of RNA conformation and dynamics. Detailed UV/vis-spectroscopic studies demonstrate that the fluorescence lifetime and the quantum yield of Çmf are directly affected by intermolecular interactions, which makes it a sensitive probe of its microenvironment. On the other hand, Çm undergoes effective fluorescence quenching in the ps-time domain. The established quenching mechanisms that are usually operational for fluorophore-nitroxide compounds, do not explain the spectroscopic data for Çm. Quantum chemical calculations revealed that the lowest excited doublet state D1, which has no equivalent in Çmf, is a key state of the ultrafast quenching mechanism. This dark state is localized on the nitroxide group and is populated via rapid internal conversion.


Amelioration of acidic soil using various renewable waste resources.
In this study, improvement of acidic soil with respect to soil pH and exchangeable cations was attempted for sample with an initial pH of approximately 5. Acidic soil was amended with various waste resources in the range of 1 to 5 wt.% including waste oyster shells (WOS), calcined oyster shells (COS), Class C fly ash (FA), and cement kiln dust (CKD) to improve soil pH and exchangeable cations. Upon treatment, the soil pH was monitored for periods up to 3 months. The exchangeable cations were measured after 1 month of curing. After a curing period of 1 month, a maize growth experiment was conducted with selected-treated samples to evaluate the effectiveness of treatment. The treatment results indicate that in order to increase the soil pH to a value of 7, 1 wt.% of WOS, 3 wt.% of FA, and 1 wt.% of CKD are required. In the case of COS, 1 wt.% was more than enough to increase the soil pH value to 7 because of COS's strong alkalinity. Moreover, the soil pH increases after a curing period of 7 days and remains virtually unchanged thereafter up to 1 month of curing. Upon treatment, the summation of cations (Ca, Mg, K, and Na) significantly increased. The growth of maize is superior in the treated samples rather than the untreated one, indicating that the amelioration of acidic soil is beneficial to plant growth, since soil pH was improved and nutrients were replenished.


Expression of the influenza virus haemagglutinin in insect cells by a baculovirus vector.
The insect baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) has played a major role in studies on the molecular biology of insect DNA viruses. Recently, this system has been effectively adapted as a highly efficient vector in insect cells for the expression of several mammalian genes. A cDNA sequence of the influenza (fowl plague) virus haemagglutinin gene has been inserted into the BamHI site of the pAc373 polyhedrin vector. Spodoptera frugiperda cells were co-transfected with this construct, pAc-HA651, and authentic AcNPV DNA. Recombinant virus was selected by adsorption of transfected cells to erythrocytes followed by serial plaque passages on S. frugiperda cells. We have determined the site of insertion of the haemagglutinin gene into the AcNPV genome by restriction enzyme cleavage and Southern blot hybridization analyses using haemagglutinin cDNA as a probe. The influenza haemagglutinin gene is located in the polyhedrin gene of AcNPV DNA. Immunofluorescent labelling, immunoprecipitation and immunoblot analyses with specific antisera revealed that S. frugiperda cells produce immune reactive haemagglutinin after infection with the recombinant virus. The haemagglutinin is expressed at the cell surface and has haemolytic capacity that has been activated by post-translational proteolytic cleavage. When chickens were immunized with S. frugiperda cells expressing haemagglutinin, they developed haemagglutinin-inhibiting and neutralizing antibodies and were protected from infection with fowl plague virus. These observations demonstrate that the haemagglutinin is processed in insect cells in a similar fashion as in fowl plaque virus-infected vertebrate cells and that it has full biological activity.


Fatty acid composition of Abies pindrow (West Himalayan fir).
The leaves of Abies pindrow, collected from Murree Hills, Punjab (Pakistan) revealed the presence of eleven fatty acids including eight saturated and three unsaturated fatty acids. They ranged from C(14) to C(24) and were detected as methyl esters by GC-MS technique. The saturated fatty acids were present in much greater proportion than unsaturated ones. Isopalmatic acid was found to be major saturated fatty acid and the oleic acid as predominant unsaturated acid. (+)-14-Methyl palmatic acid and (+)-Isosteric acid were the next higher saturated and unsaturated fatty acids respectively.


[Variations in acetylcholinesterase activity of the brain of Tenebrio molitor (Coleoptera) during metamorphosis and ecdysterone titer].
After the metamorphosis of Tenebrio molitor (Coleoptera), Acetylcholinesterase (AChE) activity per adult brain was twofold the activity of the last larval instar. During metamorphosis, from the prepupa to the adult fluctuations occurred in brain AChE activity per protein amount. These fluctuations were superimposable to the molting hormone (ecdysterone) variations in hemolymph so that each rise of ecdysterone was accompanied by an AChE peak.


[Evaluation of the MRI Findings on a Tendon Graft after the Anatomic Anterior Cruciate Ligament Reconstruction in Patients with Versus without the Application of Platelet-Rich Fibrin - Results of the Prospective Randomised Study].
PURPOSE OF THE STUDY The purpose of the prospective randomised study was to assess the benefits of the platelet-rich fibrin (PRF) application during the anatomic anterior cruciate ligament reconstruction (ACLR) for tendon graft healing. MATERIAL AND METHODS The study included 40 patients with an isolated rupture of the anterior cruciate ligament in the knee. The MRI findings on the tendon graft were evaluated at 6 months and 12 months after the surgery in a total of 33 patients, namely in 10 women/23 men (21 right knee/12 left knee), with the mean age of 29.1 years and at the mean follow-up of 15.9 weeks after the injury. The randomisation using envelopes was carried out at the beginning of arthroscopy either in the group with the PRF application (Group 1) (17 patients) or in the group without the PRF application (Group 2) (16 patients). The followed-up patients underwent the ACLR with targeting the femoral tunnel through anteromedial portal using the autografts of m. semitendinosusand m. graciliswith Tightrope femoral fixation and Inion implant in tibia. In Group 1, during the surgery PRF was applied both in the drilled tunnels (intraosseously) and directly to the surface of the tendon graft itself (intraarticular part). The Group 2 was a control group in which the same intervention was performed, however without the PRF application. In the 6 th and 12 th month after the surgery, the signal intensity of the knee and occurrence of the ACL graft failure, bone (marrow) oedema rate near the tunnels were assessed by an independent radiologist using a 3 Tesla MRI. RESULTS In the 6 th and also in the 12 th postoperative month, neither a complete, nor a partial graft failure was found in the group with PRF, whereas in Group 2 a complete or a partial graft failure was observed in 12.5% (p = 0.23). In the 6th month, a bone marrow oedema near the intraosseous parts of the graft was reported in 94.1% of patients in Group 1 and in 81.3% of patients in Group 2, while in the 12 th month it was 23.5% of evaluated patients in Group 1 and 37.5% patients in Group 2. The differences between the two groups were not statistically significant in the 6 th (p = 0.096) or in the 12 th month (p = 0.43). In the 6 th month, a fully ligamentised graft was found in 52.9% of patients in Group 1 and in 37.5% of patients in Group 2 (p = 0.061), a partially ligamentised graft was present in 47.1% of patients in Group 1 and in 50% of patients in Group 2. In the 12 th month, a fully ligamentised graft was seen in 94.1% of patients in Group 1 and in 75% of patients in Group 2 (p = 0.26) and a partially ligamentised graft was present in 5.9% of patients in Group 1 and in 12.5% of patients in Group 2. In the remaining 12.5% of patients in Group 2, the finding was assessed as a graft with no signs of ligamentisation. DISCUSSION We have not encountered a paper in the Czech or world literature which would present results of a similar study, i.e. which would evaluate the potential benefits of the platelet-rich fibrin application in the course of the anatomic anterior cruciate ligament reconstruction. Majority of studies evaluate the benefits of the application of platelet-rich plasma aimed to achieve a positive effect on the LCA graft healing. CONCLUSIONS Our study proved a higher percentage of ligamentisation and healed grafts into the bone as well as a lower occurrence of graft failure in the group with PRF compared to the group without the PRF application, namely both in the 6 th and in the 12 th month. Nonetheless, the differences between the two groups assessed at the same time were not statistically significant. Key words: anterior cruciate ligament, arthroscopy, anatomic ACL reconstruction, magnetic resonance imaging of the graft, platelet-rich fibrin, graft healing, MRI signal intensity of the graft.


Cognitive rehabilitation for memory deficits after stroke.
Memory problems are a common cognitive complaint following stroke and can potentially affect ability to complete functional activities. Cognitive rehabilitation programmes either attempt to retrain lost or poor memory functions, or teach patients strategies to cope with them.Some studies have reported positive results of cognitive rehabilitation for memory problems, but the results obtained from previous systematic reviews have been less positive and they have reported inconclusive evidence. This is an update of a Cochrane review first published in 2000 and most recently updated in 2007. To determine whether participants who have received cognitive rehabilitation for memory problems following a stroke have better outcomes than those given no treatment or a placebo control.The outcomes of interest were subjective and objective assessments of memory function, functional ability, mood, and quality of life. We considered the immediate and long-term outcomes of memory rehabilitation. We used a comprehensive electronic search strategy to identify controlled studies indexed in the Cochrane Stroke Group Trials Register (last searched 19 May 2016) and in the Cochrane Central Register of Controlled Trials (CENTRAL2016, Issue 5), MEDLINE (2005 to 7 March 2016), EMBASE 2005 to 7 March 2016), CINAHL (2005 to 5 February 2016), AMED (2005 to 7 March 2016), PsycINFO (2005 to 7 March 2016), and nine other databases and registries. Start dates for the electronic databases coincided with the last search for the previous review. We handsearched reference lists of primary studies meeting the inclusion criteria and review articles to identify further eligible studies. We selected randomised controlled trials in which cognitive rehabilitation for memory problems was compared to a control condition. We included studies where more than 75% of the participants had experienced a stroke, or if separate data were available from those with stroke in mixed aetiology studies. Two review authors independently selected trials for inclusion, which was then confirmed through group discussion. We assessed study risk of bias and extracted data. We contacted the investigators of primary studies for further information where required. We conducted data analysis and synthesis in accordance with the Cochrane Handbook for Systematic Reviews of Interventions. We performed a 'best evidence' synthesis based on the risk of bias of the primary studies included. Where there were sufficient numbers of similar outcomes, we calculated and reported standardised mean differences (SMD) using meta-analysis. We included 13 trials involving 514 participants. There was a significant effect of treatment on subjective reports of memory in the short term (standard mean difference (SMD) 0.36, 95% confidence interval (CI) 0.08 to 0.64, P = 0.01, moderate quality of evidence), but not the long term (SMD 0.31, 95% CI -0.02 to 0.64, P = 0.06, low quality of evidence). The SMD for the subjective reports of memory had small to moderate effect sizes.The results do not show any significant effect of memory rehabilitation on performance in objective memory tests, mood, functional abilities, or quality of life.No information was available on adverse events. Participants who received cognitive rehabilitation for memory problems following a stroke reported benefits from the intervention on subjective measures of memory in the short term (i.e. the first assessment point after the intervention, which was a minimum of four weeks). This effect was not, however, observed in the longer term (i.e. the second assessment point after the intervention, which was a minimum of three months). There was, therefore, limited evidence to support or refute the effectiveness of memory rehabilitation. The evidence was limited due to the poor quality of reporting in many studies, lack of consistency in the choice of outcome measures, and small sample sizes. There is a need for more robust, well-designed, adequately powered, and better-reported trials of memory rehabilitation using common standardised outcome measures.


The nit1C gene cluster of Pseudomonas pseudoalcaligenes CECT5344 involved in assimilation of nitriles is essential for growth on cyanide.
A proteomic approach was used to identify several proteins induced by cyanide in the alkaliphilic bacterium Pseudomonas pseudoalcaligenes CECT5344, two of them, NitB and NitG, encoded by genes that belong to the nit1C gene cluster. The predicted products of the nit1C gene cluster are a Fis-like σ(54) -dependent transcriptional activator (NitA), a nitrilase (NitC), an S-adenosylmethionine superfamily member (NitD), an N-acyltransferase superfamily member (NitE), a trifunctional polypeptide of the AIRS/GARS family (NitF), an NADH-dependent oxidoreductase (NitH) and two hypothetical proteins of unknown function (NitB and NitG). RT-PCR analysis suggested that nitBCDEFGH genes were co-transcribed, whereas the regulatory nitA gene was divergently transcribed. Real-time RT-PCR revealed that expression of the nitBCDEFGH genes was induced by cyanide and repressed by ammonium. The P. pseudoalcaligenes CECT5344 nit1C gene cluster was found to be involved in assimilation of free and organic cyanides (nitriles) as deduced for the inability to grow with cyanides showed by the NitA, NitB and NitC mutant strains. The wild-type strain CECT5344 showed a nitrilase activity that allows growth on cyanide or hydroxynitriles. The NitB and NitC mutants only presented low basal levels of nitrilase activity that were not enough to support growth on either free cyanide or aliphatic nitriles, suggesting that nitrilase NitC is specific and essential for cyanide and aliphatic nitriles assimilation.


Developmental kinetics, turnover, and stimulatory capacity of thymic epithelial cells.
Despite the importance of thymic stromal cells to T-cell development, relatively little is known about their biology. Here, we use single-cell analysis of stromal cells to analyze extensive changes in the number and composition of thymic stroma throughout life, revealing a surprisingly dynamic population. Phenotypic progression of thymic epithelial subsets was assessed at high resolution in young mice to provide a developmental framework. The cellular and molecular requirements of adult epithelium were studied, using various mutant mice to demonstrate new cross talk checkpoints dependent on RelB in the cortex and CD40 in the medulla. With the use of Ki67 and BrdU labeling, the turnover of thymic epithelium was found to be rapid, but then diminished on thymic involution. The various defects in stromal turnover and composition that accompanied involution were rapidly reversed following sex steroid ablation. Unexpectedly, mature cortical and medullary epithelium showed a potent capacity to stimulate naive T cells, comparable to that of thymic dendritic cells. Overall, these studies show that the thymic stroma is a surprisingly dynamic population and may have a more direct role in negative selection than previously thought.


Auxotyping of Neisseria gonorrhoeae isolated in the United Kingdom.
One hundred and twenty consecutive isolates of Neisseria gonorrhoeae from the County of Avon, UK, were examined by auxotyping and penicillin susceptibility testing. Requirements for proline (Pro), arginine (Arg), hypoxanthine (Hyp), uracil (Ura) and methionine (Met) were determined with a modified Heckels' medium. Prototrophic strains accounted for 29.9% of isolates and were a heterogeneous group as judged by penicillin susceptibility testing. The group most generally susceptible to penicillin required Arg-Hyp-Ura, and represented 27.5% of isolates. The Pro-Argo-Ura auxogroup (10% of isolates) had the narrowest range of penicillin susceptibility. Prototrophic strains had a reduced growth rate on arginine-free medium. Investigations of this phenomenon and suitable methods for auxotyping are reported.


Multiple granulomatous lung lesions in a patient with Epstein-Barr-virus-induced mononucleosis and new-onset systemic lupus erythematosus: a case report.
Granulomatous lesions are commonly encountered abnormalities in pulmonary pathology, and often pose a diagnostic challenge. We report an unusual case of granulomatous lung disease with uncommon characteristics, which developed following Epstein-Barr-virus-induced mononucleosis and new-onset systemic lupus erythematosus. We aim to highlight a diagnostic approach for the condition and to raise awareness of the possibility of it being related to the immunological reaction caused by Epstein-Barr virus infection. A 36-year-old Japanese man, who had been diagnosed with Epstein-Barr-virus-induced infectious mononucleosis, new-onset systemic lupus erythematosus, and secondary Sjögren's syndrome three weeks previously, presented to our facility with fever and diffuse pulmonary infiltrates. A computed tomography scan of the chest revealed multiple small nodules in both lungs. Fiberoptic bronchoscopy with bronchoalveolar lavage revealed lymphocytosis with predominance of T lymphocytes. A histological examination of a lung biopsy taken during video-assisted thoracic surgery showed randomly distributed tiny granulomatous lesions with infiltration of eosinophils. The differential diagnoses included hypersensitivity pneumonitis, sarcoidosis, and pulmonary involvement of Crohn's disease, systemic lupus erythematosus, and Sjögren's syndrome, but the clinical and pathological findings were not consistent with any of these. Our patient's condition did not improve; therefore, prednisolone therapy was started because of the possibility of specific immunological reactions associated with Epstein-Barr virus infection. After steroid treatment, our patient showed radiological and clinical improvement. To the best of our knowledge, this is the first case of a patient developing randomly distributed multiple granulomatous lung lesions with eosinophilic infiltrates after Epstein-Barr virus infection and systemic lupus erythematosus. On the basis of our data, we hypothesize that Epstein-Barr virus infection altered the immune response of our predisposed patient and contributed to the pathogenesis of the lung lesions. Our patient's clinical response to steroid treatment was excellent.


Psychologists' beliefs about the prevalence of childhood sexual abuse: the influence of sexual abuse history, gender, and theoretical orientation.
This study examined the influence of sexual abuse history, gender, theoretical orientation, and age on beliefs about the prevalence of childhood sexual abuse among clinical and counseling psychologists. A mail survey design was used in this study. Participants were randomly selected from the American Psychological Association membership database. There were 615 psychologists who completed self-report measures on beliefs about the prevalence of childhood sexual abuse and demographic characteristics. Overall, clinicians' scores on the prevalence of childhood sexual abuse were moderate. There were significant gender differences on beliefs, suggesting that women were more likely believe that childhood sexual abuse is a common occurrence compared to men. Multiple regression analysis indicated that clinician characteristics (history of sexual abuse, gender, and theoretical orientation) were significantly related to beliefs about the prevalence of childhood sexual abuse. However, these characteristics only accounted for a small amount of the overall variance predicting beliefs. These results suggest that clinicians do not hold extreme beliefs regarding the prevalence of childhood sexual abuse. Moreover, certain clinician characteristics are associated with their beliefs, which in turn, may impact their clinical judgment and treatment decisions. Furthermore, much of the variance was unaccounted for in the model indicating that psychologists' beliefs are complex and are not unduly influenced by their personal characteristics. Implications for clinical practice and future research are discussed.


Theoretical studies on binding and specificity mechanisms of farnesyltransferase (FTase) and geranylgeranyltransferase type-I (GGTase-I) inhibitors by molecular modeling.
Farnesyltransferase (FTase) and geranylgeranyltransferase type-I (GGTase-I) are two members of protein prenyltransferases, which play critical roles in lipid post-translational modifications. Potent inhibitors of FTase and GGTase-I have been confirmed to show favorable influence on the therapies of various diseases, such as cancers, malaria and Toxoplasmosis. However, designing highly specific inhibitors toward FTase or GGTase-I without influencing their binding affinity remains a big challenge. In this work, molecular docking, molecular dynamics (MD) simulations and MM/GBSA free energy calculations were employed to study the bindings of two highly selective inhibitors (lonafarnib and GGTI-2133) towards FTase or GGTase-I. The specificities of the studied inhibitors derived from the predicted binding free energies are consistent with the experimental data. The analysis of the energetic components illustrates that both the non-polar and polar interactions play critical roles in determining the specificity between FTase and GGTase-I. Moreover, the protein-inhibitor interaction spectra for the studied inhibitors were determined through the decomposition of the binding free energies, and the important residues for binding and specificity were highlighted. Our study provides useful information for the rational design of selective FTase or GGTase-I inhibitors.


Analysis of a beta-helical region in the p55 domain of Helicobacter pylori vacuolating toxin.
Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach and contributes to the development of gastric cancer and peptic ulcer disease. VacA, a toxin secreted by H. pylori, is comprised of two domains, designated p33 and p55. Analysis of the crystal structure of the p55 domain indicated that its structure is predominantly a right-handed parallel beta-helix, which is a characteristic of autotransporter passenger domains. Substitution mutations of specific amino acids within the p33 domain abrogate VacA activity, but thus far, it has been difficult to identify small inactivating mutations within the p55 domain. Therefore, we hypothesized that large portions of the p55 domain might be non-essential for vacuolating toxin activity. To test this hypothesis, we introduced eight deletion mutations (each corresponding to a single coil within a beta-helical segment spanning VacA amino acids 433-628) into the H. pylori chromosomal vacA gene. All eight of the mutant VacA proteins were expressed by the corresponding H. pylori mutant strains and underwent proteolytic processing to yield ~85 kDa passenger domains. Three mutant proteins (VacA Delta484-504, Delta511-536, and Delta517-544) were secreted and induced vacuolation of mammalian cells, which indicated that these beta-helical coils were dispensable for vacuolating toxin activity. One mutant protein (VacA Delta433-461) exhibited reduced vacuolating toxin activity compared to wild-type VacA. Other mutant proteins, including those containing deletions near the carboxy-terminal end of the beta-helical region (amino acids Val559-Asn628), exhibited marked defects in secretion and increased susceptibility to proteolytic cleavage by trypsin, which suggested that these proteins were misfolded. These results indicate that within the beta-helical segment of the VacA p55 domain, there are regions of plasticity that tolerate alterations without detrimental effects on protein secretion or activity, as well as a carboxy-terminal region in which similar alterations result in protein misfolding and impaired secretion. We propose that non-essential beta-helical coils and a carboxy-terminal beta-helical segment required for proper protein folding and secretion are features shared by numerous autotransporter passenger domains.


Novel interferonopathies associated with mutations in RIG-I like receptors.
Type I interferonopathies are a relatively new class of inherited autoimmune disorders associated with an inborn elevated interferon response. Activation of cytosolic receptors which recognize viral double stranded RNA including the RIG-I (retinoic acid-inducible gene I) like receptors RIG-I and MDA5 (melanoma differentiation-associated gene 5) has been shown to induce the transcription of type I interferon genes. Within recent years, with the help of next generation sequencing techniques in syndromic families, mutations in the genes encoding for RIG-I and MDA5 have been identified to cause rare diseases including Aicardi-Goutières syndrome, Systemic Lupus Erythematosus in certain individuals as well as classic and atypical Singleton-Merten syndrome. Patients carrying mono-allelic mutations in MDA5 and RIG-I show constitutive activation of the RIG-I receptors and downstream signalling associated with increased type I interferon production. Although differing in the degree of phenotypic expression and severity, the phenotype of these "novel" diseases shows a considerable overlap reflecting their common pathogenetic pathway.


What are the support needs of men with multiple sclerosis, and are they being met?
Multiple sclerosis (MS) is a chronic, degenerative condition of the central nervous system that can lead to high levels of disability and a range of psychological and social problems. Although considerable research has been conducted on the experience of living with MS and the disease's psychosocial consequences, much of this has focused on women, with the male perspective seldom considered. It is possible, therefore, that men differ from women in how they experience MS and the type of support or interventions that meet their needs. A literature review was undertaken to identify the specific support needs of men with MS, describe current support available to men with MS, and evaluate the extent to which these needs are met by current service provision. The literature reviewed suggests that the needs of men with MS are different from those of women in terms of psychological factors such as well-being and mental health and support. If interventions are to be evidence based, health-care professionals need to take into account the needs, desires, and capabilities of men with MS in the development of services. Further qualitative and quantitative research is required to address the gaps in the evidence base of support needs for men with MS.


Sensitive visual test for concave diffraction gratings.
A simple visual test for the evaluation of concave diffraction gratings is described. It is twice as sensitive as the Foucault knife edge test, from which it is derived, and has the advantage that the images are straight and free of astigmatism. It is particularly useful for gratings with high ruling frequency where the above image faults limit the utility of the Foucault test. The test can be interpreted quantitatively and can detect zonal grating space errors of as little as 0.1 A.


What have we learned in the last 20 years? A comparison of a modern era pediatric and congenital catheter ablation registry to previous pediatric ablation registries.
Since the onset of pediatric catheter ablation, the pediatric electrophysiology community has reported outcomes via various registries (PAPCA [Prospective Assessment After Pediatric Cardiac Ablation], PCAR [Pediatric Catheter Ablation Registry]). Most recently, a modern era pediatric and congenital ablation registry (MAP-IT [Multicenter Pediatric and Congenital EP Quality Initiative]) was developed for eventual incorporation into the National Cardiovascular Data Registry (NCDR) IMPACT (Improving Pediatric and Adult Congenital Treatment) registry. The purpose of this study was to describe initial findings from the MAP-IT pilot registry and to compare these findings to earlier registries. Before entering the NCDR IMPACT registry, MAP-IT was active at 12 centers (11 in the United States) between October 2014 and April 2016. All electrophysiological studies for patients younger than 21 years and for patients of all ages with structural congenital heart disease were included. We compared the acute success, fluoroscopy and procedural times, and frequency of complications between MAP-IT and the earlier registries. Acute success rates have improved from the initial PCAR registry for both accessory and slow pathway substrates. Both fluoroscopy and procedural times have significantly decreased across the time periods (fluoroscopy time 47.6 ± 40 minutes to 7.0 ± 9.2 minutes; P <.001; procedural time 257 ± 157 minutes to 166 ± 84 minutes; P <.001). Acute success rates and fluoroscopy and procedural times in pediatric ablation all have improved over the last 25 years.


Epithelioid hemangioma of brachial artery: report of a case and review of the literature.
Epithelioid hemangioma (EH) is an uncommon benign vascular lesion, also known as angioblastic lymphoid (or angiolymphoid) hyperplasia with eosinophilia, characterized by an unclear etiopathogenesis. It usually affects young to middle-aged adults and develops in the head and neck region, as painless cutaneous or subcutaneous reddish papules or nodules. Large vessels involvement is extremely rare, and to date only two cases affecting the brachial artery have been cited in literature. In this report we present a further case of EH of the brachial artery and review the pertinent literature.


New South Wales group pathology services--the Southpath model.
There has been significant rationalisation of public pathology service provision in New South Wales through the formation of group pathology services. These services are a fundamental change in the method of service organisation and delivery and little has been reported detailing their organisational and financial structures. Southpath, the group pathology service of the Southern Sydney Area Health Service, has improved service delivery, improved productivity by 18.2 per cent and reduced pathology expenditure per admission by 19.2 per cent. This paper describes the organisation of Southpath as knowledge of the organisational structures and performance of the group pathology service is necessary for the evaluation of this change in the method of service delivery and is essential to the current debate on the value of public pathology services.


Do all trauma patients benefit from tranexamic acid?
This study tested the hypothesis that early routine use of tranexamic acid (TXA) reduces mortality in a subset of the most critically injured trauma intensive care unit patients. Consecutive trauma patients (n = 1,217) who required emergency surgery (OR) and/or transfusions from August 2009 to January 2013 were reviewed. At surgeon discretion, TXA was administered at a median of 97 minutes (1-g bolus then 1-g over 8 hours) to 150 patients deemed high risk for hemorrhagic death. With the use of propensity scores based on age, sex, traumatic brain injury (TBI), mechanism of injury, systolic blood pressure, transfusion requirements, and Injury Severity Score (ISS), these patients were matched to 150 non-TXA patients. The study population was 43 years old, 86% male, 54% penetrating mechanism of injury, 25% TBI, 28 ISS, with 22% mortality. OR was required in 78% at 86 minutes, transfusion was required in 97% at 36 minutes, and 75% received both. For TXA versus no TXA, more packed red blood cells and total fluid were required, and mortality was 27% versus 17% (all p < 0.05). The effects of TXA were similar in those with or without TBI, although ISS, fluid, and mortality were all higher in the TBI group. Mortality associated with TXA was influenced by the timing of administration (p < 0.05), but any benefit was eliminated in those who required more than 2,000-mL packed red blood cells, who presented with systolic blood pressure of less than 120 mm Hg or who required OR (all p < 0.05). For the highest injury acuity patients, TXA was associated with increased, rather than reduced, mortality, no matter what time it was administered. This lack of benefit can probably be attributed to the rapid availability of fluids and emergency OR at this trauma center. Prospective studies are needed to further identify conditions that may override the benefits from TXA. Therapeutic study, level IV.


Bilateral childhood visual impairment: child and parent concerns.
To identify specific health-related quality of life and visual function concerns affecting children with bilateral visual impairment as expressed by children or one of their parents (proxy) and concerns affecting the parents themselves. A total of 37 children <16 years of age with visual impairment (visual acuity worse than 20/70 in the better eye) and one parent for each child were prospectively enrolled. Semistructured individual interviews were performed with children 5-15 years of age (n = 16) and with one parent for each child (ages 0-15 years, N = 37). Interview transcripts were analyzed using NVivo software. Categories of concern were identified from both child and parent interviews, from which broad themes were identified. The frequencies of the themes and specific categories of concerns were calculated. Regarding the child's experience, categories of concern were grouped into 6 themes: visual function (expressed by 13 of 16 children [81%] and 33 of 37 parents [89%]), treatment (63% and 54%), emotions (50% and 68%), social (50% and 70%), physical discomfort (50% and 22%), and worry (38% and 8%). Concerns expressed regarding the parents' own experience were grouped into 5 themes: worry (100%), compensate-adjust for condition (89%), treatment (84%), emotions (81%), and affects family (46%). Individual interviews identified a wide spectrum of concerns in children with visual impairment and their parents, affecting functional, emotional, social and physical domains. Specific concerns will be used to develop patient-derived questionnaires for quantifying the effects of visual impairment on children and parents in everyday life.


High-dose cyclophosphamide without stem cell rescue in immune-mediated necrotizing myopathies.
To describe the experience managing treatment-refractory immune-mediated necrotizing myopathies (IMNM) with high-dose cyclophosphamide (HiCy) therapy. Five patients with severe refractory IMNM who were treated with HiCy without stem cell rescue were identified. Their medical records were reviewed to assess demographic, clinical, and histologic characteristics as well as response to therapy. Three patients with anti-signal recognition particle (SRP) and 2 patients with anti-HMG-CoA reductase autoantibodies were included. The mean follow-up time after HiCy therapy was 37 ± 28 months. Two patients demonstrated substantial response, evidenced by improved muscle strength and decreased muscle enzymes after HiCy therapy; both of these patients were anti-SRP positive. Four patients experienced febrile neutropenia after HiCy therapy, one of which required a prolonged intensive care unit stay for infectious complications, from which they eventually recovered. These data suggest that HiCy therapy without stem cell rescue may be considered as an alternative for the treatment of refractory IMNM.


The tryptophan synthase-encoding trpB gene of Aspergillus nidulans is regulated by the cross-pathway control system.
The tryptophan synthase-encoding gene, trpB, of Aspergillus nidulans was cloned and characterized. It was mapped to chromosome I, between the gene medA, which is required for sexual and asexual development, and an ORF encoding a protein with significant similarity to subunit B of vacuolar ATP synthases. The 5' untranslated region was found to be at least 142 nucleotides (nt) long, the poly(A) addition site was localized at position + 216 relative to the stop codon by sequencing of several independent cDNA clones. The trpB gene contains two exons separated by an intron of 105 nt, which is located close to the 5' end of the ORF. Directly upstream of the transcriptional start site, one well conserved potential binding site for the cross-pathway control transcriptional activator CPCA was found. The level of trpB transcript was shown to be regulated by cross-pathway control. A knockout mutant for trpB displays tryptophan auxotrophy, no trpB transcript is detectable, and development is perturbed to an extent that is dependent on the amount of tryptophan added to the medium. The trpB gene encodes a protein of 723 amino acids, with a calculated molecular weight of 77.6 kDa. The deduced amino acid sequence shows 72.6% similarity to the tryptophan synthase of Neurospora crassa. Most amino acid residues essential for catalytic activity in the tryptophan synthase of Salmonella typhimurium are conserved. The linker region joining the two domains of the enzyme is 13 residues longer than the longest connector found so far in tryptophan synthases from fungi.


Comparison of vertical forces during root canal filling with three different obturation techniques.
The aim of this study was to examine and compare vertical forces exerted during root canal obturation with the cold lateral condensation technique, Thermafil technique and ProTaper guttapercha. Fourty-five single-rooted permanent teeth were used in the study. All specimens were instrumented using the ProTaper rotating technique and were randomly divided into three experimental groups (n = 15 per group). In the first group, root canals were obturated using the cold lateral condensation technique. In the second group, the Thermafil technique was used to obturate root canals. In the last group, a ProTaper gutta-percha of the same taper as the instrumented root canals was used for root canal obturation. Vertical forces were measured using the device developed for simulation of endodontic treatment. The results showed a statistically significant difference (p = 0.0001) for vertical forces when cold lateral condensation obturation technique was used in comparison to other techniques. No statistically significant difference was found for vertical forces during obturation with Thermafil and ProTaper gutta-percha (p = 0.16). The cold lateral condensation technique exerted higher vertical forces in comparison to the Thermafil and ProTaper obturation techniques.


Subtle renal duplication as an unrecognized cause of childhood incontinence: diagnosis by magnetic resonance urography.
Urinary incontinence in young girls who have been toilet trained may be due to an ectopic ureter inserting below the urinary sphincter. This diagnosis is frequently delayed, is psychologically distressing, and may be missed at physical examination. Findings at ultrasound evaluation may be subtle and imaging with computed tomography or intravenous urography exposes young patients to ionizing radiation. We report two cases of girls with urinary incontinence where magnetic resonance (MR) urography revealed subtle renal duplication which implied the presence of an ectopic duplicated ureter with infrasphincteric insertion. These cases stress the importance of examining the kidneys, rather than the perineum, at MR, ultrasound and intravenous urogram evaluation, and show the value of MR urography as a safe alternative to computed tomography and intravenous urography for making this diagnosis.


Body composition and performance in cross-country skiing.
The purpose of this study was to investigate the relationships between body composition and performance in cross-country skiing. Ten male college-aged elite cross-country skiers (17.9 yrs; S 1.0 yrs) participated in a 5.6-km cross-country skiing time trial and in dual energy X-ray absorptiometry (DXA, Lunar DPX-L, Madison, WI, USA) body composition measurements. A differential global positioning system (dGPS, GPS 12 CX, Garmin Int. Inc., Olathe, KS, USA; RXMAR 2, Aztec SA, Strasbourg, France) was used to compute speed in different sections of the course. Spearman correlation analyses were applied. Total body weight and absolute lean body mass were significantly related to final time (r = - 0.721; p < 0.05 and - 0.830; p < 0.01, respectively). Absolute lean arm mass (kg) was negatively correlated to final time (r = - 0.648; p < 0.05) and the relative lean arm mass was significantly related to speed mainly in uphill sections (r = 0.636 to 0.867; p < 0.05 to p < 0.01). We suggest that large amounts of lean body mass, especially in the arms, seem to be of great importance for cross-country skiing performance.


Plasticity of growth and sylleptic branchiness in two poplar families grown at three sites across Europe.
Two hybrid poplar maternal half-sib families, resulting from controlled crosses of the female parent Populus deltoides 'S9-2' with P. nigra 'Ghoy' and P. trichocarpa 'V24', were grown at three sites: northern Italy, central France and southern England. Juvenile stem growth traits (height, circumference and volume) and sylleptic branchiness (number of branches, density of branches per unit of stem height, percentage of the stem carrying branches and distance of the highest sylleptic branch to the top of the stem) were measured on 1-year-old shoots. Our general objectives were to determine the degree to which the expression of stem growth and syllepsis and the relationships between them are affected by environmental conditions and to evaluate the efficiency of indirect selection for stem growth using branching traits as secondary criteria. The performance of both families differed significantly within and between sites. Pronounced heterosis was observed and highly significant genotype x environment interactions were found for all traits across the sites. Syllepsis showed more marked genetic variation and plasticity than stem growth traits. Relationships between sylleptic branchiness and stem growth depended on environmental conditions. Heritability values at the individual level ranged between 0.09 and 0.59, but genetic gain in stem volume was not significantly improved when selection was based on sylleptic branch characteristics. However, despite strong phenotypic plasticity among sites, genotypic ranking among the sites was relatively stable.


Emerging investigator series: the role of chemical properties in human exposure to environmental chemicals.
One of the ultimate goals of environmental exposure science is to mechanistically understand how chemical properties and human behavior interactively determine human exposure to the wide spectrum of chemicals present in the environment. This comprehensive review assembles state-of-the-art knowledge of the role of partitioning, dissociation, mass transfer, and reactive properties in human contact with and absorption of organic chemicals via oral, dermal, and respiratory routes. Existing studies have revealed that chemicals with different properties vary greatly in mass distribution and occurrence among multiple exposure media, resulting in distinct patterns of human intake from the environment. On the other hand, these chemicals encounter different levels of resistance in the passage of intestinal, dermal, and pulmonary absorption barriers and demonstrate different levels of bioavailability, due to the selectivity of biochemical, anatomical and physiological structures of these absorption barriers. Moving forward, the research community needs to gain more in-depth mechanistic insights into the complex processes in human exposure, advance the technique to better characterize and predict chemical properties, generate and leverage experimental data for a more diverse range of chemicals, and describe better the interactions between chemical properties and human behavior.


Insulin-like growth factor I mRNA levels are developmentally regulated in specific regions of the rat brain.
The expression of mRNAs encoding insulin-like growth factor I (IGF-I) and the IGF-I receptor in the developing rat brain from embryonic day 16 to postnatal day 82 was analyzed using solution hybridization-RNase protection assays. Four distinct developmental patterns in the steady-state levels of IGF-I mRNA were seen. Specifically, the olfactory bulb showed a high perinatal level of IGF-I mRNA which declined dramatically by postnatal day 8. In contrast, cerebral cortex displayed maximal levels of IGF-I mRNA at postnatal day 8 and 13, which subsequently declined to adult levels (P82). A third developmental pattern was seen in the hypothalamus, where IGF-I mRNA increased from E16 up to postnatal day 3 and remained elevated thereafter. Finally, IGF-I mRNA levels in brainstem and cerebellum remained unchanged throughout the time period studied. We conclude that there are specific regional patterns of IGF-I gene expression in the developing rat brain. In contrast, IGF-I receptor gene expression did not exhibit any region-specific developmental changes. The developmental patterns of IGF-I gene expression seen in this study further substantiate the potential role of IGF-I in normal brain development.


Case Report: Neuromyelitis Optica After Treatment of Uveal Melanoma With Nivolumab and Ipilimumab.
The development of immune checkpoint inhibitors (ICIs) has greatly improved survival of patients with advanced malignancies. ICIs can cause immune-related adverse effects (irAEs) involving any organ. Neurological irAEs are infrequent and have mostly been reported in patients with melanoma. We describe the case of a 57-year-old male with right eye uveal melanoma, gene expression profile (class 2), and PRAME (preferentially expressed antigen in melanoma) positivity, who received plaque brachytherapy with Iodine-125 for 4 days with subsequent adjuvant ICIs (immune checkpoint inhibitors), nivolumab and ipilimumab. 18 weeks after discontinuation of immunotherapy, the patient presented with acute onset of left-sided headaches, pain with eye movements, and vision loss. The patient was tested positive for serum anti-aquaporin-4 antibody (AQP4-Ab) and was diagnosed with neuromyelitis optica spectrum disorder (NMOSD). Subsequently, he was treated with 5 days of intravenous methylprednisolone followed by an oral prednisone taper over 10 weeks, with improvement in symptoms. We report a unique case of neuromyelitis optica spectrum disorder (NMOSD) following treatment with ICIs. To our best knowledge, this is the third reported case in English literature of NMOSD following ICI therapy and the first reported case of NMOSD caused by ICI treatment in uveal melanoma.


Phage typing of Staphylococcus saprophyticus.
This study included 502 staphylococcus strains; Staphylococcus saprophyticus (297 strains) S. cohnii (47), S. xylosus (10), S. epidermidis (67) and S. aureus (81). Mitomycin C induction was performed on 100 isolates of S. saprophyticus and all induced strains were reacted with each other. Twenty-six strains proved to be lysogenic. Phages were propagated and titrated. With 12 of the phages there were three frequent associations, named lytic groups A, B and C, which included 75% of all typable strains. Typability of the system was 45% and reproducibility was between 94.2% and 100%. Phages did not lyse S. aureus and S. epidermidis strains, but they lysed S. saprophyticus and only rare strains of other novobiocin resistant species. Effective S. saprophyticus typing serves ecological purposes and tracing the origin of urinary strains from the skin or mucous membranes. Phage typing in association with plasmid profiling previously described, are anticipated as complementary methods with strong discriminatory power for differentiating among S. saprophyticus strains.


Auditory neuropathy and a mitochondrial disorder in a child: case study.
A child was referred for an audiologic evaluation, to include auditory brainstem evoked response testing, due to inconsistent responses to sound and delayed speech and language development. Results were characteristic of auditory neuropathy. In view of subsequent decline in motor function, a genetics evaluation was conducted, revealing a mitochondrial disorder. A brief overview of mitochondrial genetics in association with hearing loss is presented. The patient's audiologic profile is described and the implications for management are discussed.


Synaptic functions in rat sympathetic neurons in microcultures. III. A Purinergic effect on cardiac myocytes.
In the first two of this series of papers, a sensitive microculture procedure was used to show that rat sympathetic neurons grown singly on small islands of heart cells release norepinephrine (NE) and/or acetylcholine (ACh). We report here the release of a third transmitter in response to stimulation of these neurons. This agent was recognized by its effect on the cocultured cardiac myocytes: an inhibition of beating or a hyperpolarization that, in contrast to cholinergic inhibition, was unaffected by atropine (up to 5 microM). Evidence described here indicates that this agent was primarily adenosine (or a closely related compound): the atropine-resistant myocyte inhibition was antagonized by adenosine-receptor blockers [8-phenyltheophylline, theophylline, 7-(2-chloroethyl) theophylline] and was attenuated by an enzyme (adenosine deaminase) that hydrolyzes adenosine to pharmacologically inactive inosine. Many of the neurons, whether initially dissociated from ganglia of newborn or adult rats, evoked this purinergic response, almost always in combination with adrenergic and cholinergic responses. In a few cases it was the only detectable response. The relative strength of the adrenergic, cholinergic, and purinergic responses varied widely from neuron to neuron, suggesting that the adrenergic and purinergic or the cholinergic and purinergic agents were not stored at constant stoichiometric ratios.


Rhythms of Transcription in Field-Grown Sugarcane Are Highly Organ Specific.
Circadian clocks improve plant fitness in a rhythmic environment. As each cell has its own circadian clock, we hypothesized that sets of cells with different functions would have distinct rhythmic behaviour. To test this, we investigated whether different organs in field-grown sugarcane follow the same rhythms in transcription. We assayed the transcriptomes of three organs during a day: leaf, a source organ; internodes 1 and 2, sink organs focused on cell division and elongation; and internode 5, a sink organ focused on sucrose storage. The leaf had twice as many rhythmic transcripts (>68%) as internodes, and the rhythmic transcriptomes of the internodes were more like each other than to those of the leaves. Among the transcripts expressed in all organs, only 7.4% showed the same rhythmic pattern. Surprisingly, the central oscillators of these organs - the networks that generate circadian rhythms - had similar dynamics, albeit with different amplitudes. The differences in rhythmic transcriptomes probably arise from amplitude differences in tissue-specific circadian clocks and different sensitivities to environmental cues, highlighted by the sampling under field conditions. The vast differences suggest that we must study tissue-specific circadian clocks in order to understand how the circadian clock increases the fitness of the whole plant.


[Effect of wenxin granule on plasma BNP and HRV of acute coronary syndrome patients].
To observe the effect of Wenxin Granule (WG) on brain natriuretic peptide (BNP) and heart rate variability (HRV) of acute coronary syndrome (ACS) patients. Totally 65 ACS patients were randomly assigned to the treatment group (35 cases) and the control group (30 cases). All patients were treated with routine therapies such as angiotensin conversing enzyme inhibitors (ACEI) and metoprolol. Those in the treatment group took WG, 9 g each time, three times daily. All were treated for 90 days. Plasma samples of BNP and HRV were determined before treatment and after treatment. There was no statistical difference in pre-treatment plasma BNP (P > 0.05). Plasma BNP significantly decreased after treatment in the two groups when compared with before treatment (P < 0.05). The decrease was more obvious in the treatment group (P < 0.05). There was no statistical difference in pre-treatment HRV (P > 0.05). Compared with before treatment in the same group, RMSSD, PNN50%, and high frequency (HF) obviously increased, while low frequency (LF) and LF/HF ratio significantly decreased in the two groups, showing statistical difference (P < 0.05). The aforesaid indices were obviously better in the treatment group than in the control group (P < 0.05). Additional administration of WG could improve short-term clinical prognosis by down-regulating plasma BNP level (via improving myocardial ischemia) and modulating HRV.


Distribution of arrestin-like protein and beta-subunit of GTP-binding proteins in quail choroid plexuses.
Monoclonal antibodies (Mabs) directed against retinal arrestin (S-antigen) were used to detect and characterize this protein in choroid plexus (CP) of quails maintained during eight days, either under long-day photoperiods or in constant darkness. Immunocytochemistry and Western blotting confirmed the presence and the distribution of an arrestin-like protein in quail CP. Arrestin-like immunoreactivities in CP were compared with those obtained with Mabs to beta 36-subunit of G proteins (G beta), alpha-subunit of transducin and rhodopsin. Rhodopsin-like and transducin-like proteins could not be detected in choroidal cells, whereas intense positive reactions were observed with anti-G beta and anti-arrestin Mabs. The strongest immunoreactivities were found in choroidal ependymocytes of the lateral and IIIrd ventricles. In CP epithelial cells lining the IVth ventricle, very weak or no immunoreactivity could be detected with Mabs to arrestin, while Mab against G beta subunit always provided a positive reaction. In quails maintained in constant darkness, arrestin- and G beta-immunoreactivities of CP epithelial cells displayed changes in cellular distribution and intensity (decrease or disappearance of the immunoreactions). The strong arrestin-like immunoreaction located in the apical region of ependymocytes suggests the preferential association of the protein with choroidal microvilli and a possible role in cerebrospinal fluid production assumed by CP cells.