Protein O-mannosyltransferases participate in ER protein quality control.
In eukaryotic cells, proteins enter the secretory pathway at the endoplasmic reticulum (ER) as linear polypeptides and fold after translocation across or insertion into the membrane. If correct folding fails, many proteins are O-mannosylated inside the ER by an O-mannosyltransferase, the Pmt1p-Pmt2p complex. The consequences of this modification are controversial and the cellular role of the Pmt1p-Pmt2p complex in this respect is unclear. Here, we have identified the binding partners of yeast Pmt1p and Pmt2p. These include ER chaperones involved in oxidative protein folding; the Hrd1p complex, which is involved in ER-associated protein degradation (ERAD); and the p24 protein complex involved in ER export. The results suggest that the Pmt1p-Pmt2p complex participates in these processes. We tested this assumption in a functional assay and found that whereas the Pmt1p-Pmt2p complex promotes fast ER export of the GPI-anchored protein Gas1p, it retains the misfolded version Gas1*p and targets it to the Hrd1p complex for subsequent degradation. Our results reveal previously unknown cellular roles of the Pmt1p-Pmt2p complex in connection with the ERAD machinery and show its participation in ER protein quality control.


Effectiveness of sonohysterography in infertile patient work-up: a comparison with transvaginal ultrasonography and hysteroscopy.
The aim of this comparative study was to evaluate the accuracy of transvaginal ultrasonography (TVU), sonohysterography (SHG) with sterile saline solution compared to hysteroscopy in the diagnosis of intrauterine pathology in a population of infertile patients before an in vitro fertilization (IVF) program. 98 infertile patients with a mean age of 33.9 years (range 27-41) underwent TVU assessment and SHG with sterile saline solution immediately followed by hysteroscopy. The clinical findings obtained by TVU, SHG and hysteroscopy were compared. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of TVU and SHG compared to hysteroscopy were calculated. The TVU sensitivity and specificity compared with hysteroscopy were 91 and 83% respectively. Using TVU, a 9.2% false positive rate (9 cases) and a 5.1% false negative rate (5 cases) were detected compared to hysteroscopy. The TVU PPV and NPV were 85.4 and 90% respectively. SHG yielded better results: sensitivity and specificity when compared to hysteroscopy were 98 and 94% respectively. The SHG PPV and NPV were 95 and 98% respectively. Accuracy of SHG was significantly better than TVU considering all intrauterine pathologies and polyps. In experienced hands, SHG is an easy, safe, and well-tolerated alternative to diagnostic hysteroscopy in the initial evaluation of uterine cavity infertile patients.


Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithelium of light-exposed mice.
To elucidate the influences of light exposure on the retinal pigment epithelium (RPE) in vivo that may be involved in the pathogenesis of AMD. Six- to 7-week-old BALB/c mice were exposed to light at 2000 lux for 3 hours. Flat-mount RPE samples were immunostained with anti-ZO-1 antibody for evaluating tight junction, anti-N-cadherin, and anti-β-catenin antibodies for adherens junction, and stained with phalloidin for actin cytoskeleton. The reactive oxygen species (ROS) level was measured using DCFH-DA; Rho-associated coiled-coil forming kinase (ROCK) activity was by ELISA. Cytokine expression was analyzed by real-time RT-PCR and/or ELISA in the RPE-choroid, and macrophage recruitment was by real-time RT-PCR and immunohistochemistry. Either an antioxidant, N-Acetyl-L-cysteine (NAC), or a ROCK inhibitor, Y-27632, were administered to analyze the roles of ROS and ROCK activation, respectively. Light exposure disrupted staining patterns of tight junctions, adherens junctions, and actin cytoskeleton in the RPE, where ROS was elevated. However, NAC treatment avoided the RPE changes, reducing ROS. ROCK activity increased after light exposure was suppressed by NAC, and the structural disruptions were suppressed by Y-27632. The levels of MCP-1, CCL11, and IL-6 increased after light exposure were suppressed by NAC. Light-induced MCP-1 and IL-6 were suppressed by Y-27632. Macrophage recruitment after light exposure was also suppressed either by NAC or Y-27632. Light exposure induced ROS and Rho/ROCK activation, which caused disruption of cell-cell junctions (tight junctions and adherens junctions) and actin cytoskeleton, the RPE's barrier structure, and induced AMD-associated pathological changes in the RPE-choroid.


miR-181c promotes proliferation via suppressing PTEN expression in inflammatory breast cancer.
Primary inflammatory breast cancer (IBC) accounts for ~6% of new breast cancer cases. Even with multimodality treatment, the 5-year disease-free survival is <45%, thus making IBC the most deadly form of locally advanced breast cancer. Better understanding of the pathogenesis of IBC is essential to the design of effective therapy. We found that miR-181c was upregulated in IBC, implying that it could be a useful prognostic marker for IBC and a novel therapeutic target for the intervention of disease. Elucidating why the gene is overexpressed and how to downregulate it will help us to further understand the pathogenesis and progression of the disease and offer new targets for therapies. In this study, we showed that miR-181c as an oncogene promoted proliferation and it inhibited PTEN protein expression by targeting 3'-UTR of PTEN mRNA in IBC SUM149 cells. Moreover, PTEN was not only downregulated in IBC, but also inhibited proliferation in SUM149 cells and introduction of PTEN cDNA lacking the predicted sites of 3'-UTR abrogated miR-181c cellular function, suggesting that miR-181c inhibited proliferation by downregulating PTEN expression in IBC. Thus, targeting miR-181c and restoration of PTEN can be used in conjunction with other therapies to prevent progression of IBC.


Persistent subthreshold voltage-dependent cation single channels in suprachiasmatic nucleus neurons.
The hypothalamic suprachiasmatic nucleus (SCN) contains the primary circadian pacemaker in mammals, and transmits circadian signals by diurnal modulation of neuronal firing frequency. The ionic mechanisms underlying the circadian regulation of firing frequency are unknown, but may involve changes in membrane potential and voltage-gated ion channels. Here we describe novel tetrodotoxin- and nifedipine-resistant subthreshold, voltage-dependent cation (SVC) channels that are active at resting potential of SCN neurons and increase their open probability (P(o)) with membrane depolarization. The increased P(o) reflects changes in the kinetics of the slow component of the channel closed-time, but not the channel open-time or fast closed-time. This study provides a background for investigation of the possible role of SVC channels in regulation of circadian oscillations of membrane excitability in SCN neurons.


Commodity risk assessment of Corylus avellana and Corylus colurna plants from Serbia.
The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers the plant health risks posed by the two following hazelnut commodities to be imported from Serbia. 1. Bare rooted plants: 1- to 3-year-old plants of Corylus avellana or C. avellana grafted on C. colurna, without leaves. 2. Plants in pots: 2-year-old plants of C. avellana, without leaves. The assessment was performed by taking into account the available scientific information, including the technical information provided by Serbia. The relevance of any pest for this Opinion was based on evidence following defined criteria. One EU quarantine pest, i.e. Flavescence dorée phytoplasma, fulfilled all relevant criteria and was selected for further evaluation. For this pathogen, the risk mitigation measures proposed in the Technical Dossier from Serbia were evaluated separately for bare rooted plants and for plants in pots, taking into account the possible limiting factors. For the selected pathogen, an expert judgement was given on the likelihood of pest freedom of plants for planting, for both commodities taken together, considering the risk mitigation measures, including uncertainties associated with the assessment. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9,837 and 10,000 bare rooted plants and plants in pots per 10,000 would be free of Flavescence dorée phytoplasma.


Chronic thromboembolic pulmonary hypertension.
Pulmonary arterial hypertension is a severe disease that has been ignored for a long time. However, over the past 20 yrs chest physicians, cardiologists and thoracic surgeons have shown increasing interest in this disease because of the development of new therapies, that have improved both the outcome and quality of life of patients, including pulmonary transplantation and prostacyclin therapy. Chronic thromboembolic pulmonary arterial hypertension (CTEPH) can be cured surgically through a complex surgical procedure: the pulmonary thromboendarterectomy. Pulmonary thromboendarterectomy is performed under hypothermia and total circulatory arrest. Due to clinically evident acute-pulmonary embolism episodes being absent in > 50% of patients, the diagnosis of CTEPH can be difficult. Lung scintiscan showing segmental mismatched perfusion defects is the best diagnostic tool to detect CTEPH. Pulmonary angiography confirms the diagnosis and determines the feasibility of endarterectomy according to the location of the disease, proximal versus distal. The technique of angiography must be perfect with the whole arterial tree captured on the same picture for each lung. The lesions must start at the level of the pulmonary artery trunk, or at the level of the lobar arteries, in order to find a plan for the endarterectomy. When the haemodynamic gravity corresponds to the degree of obliteration, pulmonary thromboendarterectomy can be performed with minimal perioperative mortality, providing definitive, excellent functional results in almost all cases.


Interleukin-22 and acute pancreatitis: A review.
Acute pancreatitis (AP) is one of the most common gastrointestinal diseases, and it is divided into 3 types according to its severity:mild acute pancreatitis, moderately severe acute pancreatitis, and severe acute pancreatitis. The mortality in severe acute pancreatitis is approximately 15% to 30% due to multiorgan dysfunction and the lack of specific treatment. Interleukin-22 (IL-22) is a member of the Interleukin-10 family, and it can activate several downstream signaling pathways by binding to its receptor complex, thus it is involved in cell differentiation, proliferation, and apoptosis. Some studies have reported the elevated level of IL-22 in patients with AP, which suggests IL-22 may be involved in the pathogenesis of AP. And many studies have shown that IL-22 had a protective effect against AP. This article reviews the characteristics and mechanism of IL-22 and its role in AP to provide insight into the treatment of AP.


How US institutional review boards decide when researchers need to translate studies.
Informed consent is crucial in research, but potential participants may not all speak the same language, posing questions that have not been examined concerning decisions by institutional review boards (IRBs) and research ethics committees' (RECs) about the need for researchers to translate consent forms and other study materials. Sixty US IRBs (every fourth one in the list of the top 240 institutions by The National Institutes of Health funding) were contacted, and leaders (eg, chairs) from 34 (response rate=57%) and an additional 12 members and administrators were interviewed. IRBs face a range of problems about translation of informed consent documents, questionnaires and manuals-what, when and how to translate (eg, for how many or what proportion of potential subjects), why to do so and how to decide. Difficulties can arise about translation of specific words and of broader cultural concepts regarding processes of informed consent and research, especially in the developing world. In these decisions, IRBs weigh the need for autonomy (through informed consent) and justice (to ensure fair distribution of benefits and burdens of research) against practical concerns about costs to researchers. At times IRBs may have to compromise between these competing goals. These data, the first to examine when and how IRBs/RECs require researchers to translate materials, thus highlight a range of problems with which these committees struggle, suggesting a need for further normative and empirical investigation of these domains, and consideration of guidelines to help IRBs deal with these tensions.


Social structure learning in human anterior insula.
Humans form social coalitions in every society, yet we know little about how we learn and represent social group boundaries. Here we derive predictions from a computational model of latent structure learning to move beyond explicit category labels and interpersonal, or dyadic, similarity as the sole inputs to social group representations. Using a model-based analysis of functional neuroimaging data, we find that separate areas correlate with dyadic similarity and latent structure learning. Trial-by-trial estimates of 'allyship' based on dyadic similarity between participants and each agent recruited medial prefrontal cortex/pregenual anterior cingulate (pgACC). Latent social group structure-based allyship estimates, in contrast, recruited right anterior insula (rAI). Variability in the brain signal from rAI improved prediction of variability in ally-choice behavior, whereas variability from the pgACC did not. These results provide novel insights into the psychological and neural mechanisms by which people learn to distinguish 'us' from 'them.'


Catalytic, Enantioselective β-Protonation through a Cooperative Activation Strategy.
The NHC-catalyzed transformation of unsaturated aldehydes into saturated esters through an organocatalytic homoenolate process has been thoroughly studied. Leveraging a unique "Umpolung"-mediated β-protonation, this process has evolved from a test bed for homoenolate reactivity to a broader platform for asymmetric catalysis. Inspired by our success in using the β-protonation process to generate enals from ynals with good E/Z selectivity, our early studies found that an asymmetric variation of this reaction was not only feasible, but also adaptable to a kinetic resolution of secondary alcohols through NHC-catalyzed acylation. In-depth analysis of this process determined that careful catalyst and solvent pairing is critical for optimal yield and selectivity; proper choice of nonpolar solvent provided improved yield through suppression of an oxidative side reaction, while employment of a cooperative catalytic approach through inclusion of a hydrogen bond donor cocatalyst significantly improved enantioselectivity.


Surface localization of Pseudomonas aeruginosa outer membrane porin protein F by using monoclonal antibodies.
Hybridomas secreting highly specific monoclonal antibodies against porin protein F of Pseudomonas aeruginosa were isolated. These antibodies interacted with protein F in outer membranes isolated from strains representing the 17 serotypes of P. aeruginosa and from another 15 clinical isolates from patients with cystic fibrosis. The cell surface localization of antigenic sites on protein F was shown by indirect immunofluorescent techniques with these monoclonal antibodies. No fluorescence was observed on a protein F-deficient strain H283 of P. aeruginosa. Another monoclonal antibody specific for outer membrane lipoprotein H2 of P. aeruginosa showed no fluorescence on intact, wild-type bacterial cells, but was able to interact with a rough, LPS-deficient mutant.


Comparison of two methods and three end points in determination of in vitro activity of micafungin against Aspergillus spp.
We investigated the in vitro activity of micafungin against clinical Aspergillus isolates (n = 37) (Aspergillusfumigatus [n = 21], Aspergillusflavus [n = 14], and Aspergillus niger [n = 2]) by using NCCLS M38A microdilution and an investigational disk diffusion assay. Microdilution assay results were evaluated by using the end points of a MIC-2 (measured in micrograms per milliliter) and minimum effective concentration (MEC, measured in micrograms per milliliter; the lowest concentration of micafungin that produces short and aberrant hyphal branchings microscopically). Disk diffusion results were interpreted by measuring the zone(s) of inhibition (ZOI, measured in millimeters). Micafungin proved to be similarly active against all Aspergillus species tested. At 24 h, MIC-2s and MECs were identical. At 48 h, however, MIC-2s increased unpredictably, leading to the loss of a consistent correlation between the two end points. MECs and ZOI remained consistent and correlated at both reading times, suggesting their use as relevant end points in susceptibility testing of micafungin against Aspergillus: All Aspergillus isolates yielded intrazonal growth on disk diffusion agar plates. The intrazonal colonies contained short, aberrant hyphal branchings microscopically. The in vivo significance of these findings remains to be further investigated.


Association of a novel endometrial cancer biomarker panel with prognostic risk, platinum insensitivity, and targetable therapeutic options.
During the past decade, the age-adjusted mortality rate for endometrial cancer (EC) increased 1.9% annually with TP53 mutant (TP53-mu) EC disproportionally represented in advanced disease and deaths. Therefore, we aimed to assess pivotal molecular parameters that differentiate clinical outcomes in high- and low-risk EC. Using the Cancer Genome Atlas, we analyzed EC specimens with available DNA sequences and quantitative gene-specific RNA expression data. After polymerase ɛ (POLE)-mutant specimens were excluded, differential gene-specific mutations and mRNA expressions were annotated and integrated. Consequent to TP53-mu failure to induce p21, derepression of multiple oncogenes harboring promoter p21 repressive sites was observed, including CCNA2 and FOXM1 (P < .001 compared with TP53 wild type [TP53-wt]). TP53-wt EC with high CCNA2 expression (CCNA2-H) had a targeted transcriptomic profile similar to that of TP53-mu EC, suggesting CCNA2 is a seminal determinant for both TP53-wt and TP53-mu EC. CCNA2 enhances E2F1 function, upregulating FOXM1 and CIP2A, as observed in TP53-mu and CCNA2-H TP53-wt EC (P < .001). CIP2A inhibits protein phosphatase 2A, leading to AKT inactivation of GSK3β and restricted oncoprotein degradation; PPP2R1A and FBXW7 mutations yield similar results. Upregulation of FOXM1 and failed degradation of FOXM1 is evidenced by marked upregulation of multiple homologous recombination genes (P < .001). Integrating these molecular aberrations generated a molecular biomarker panel with significant prognostic discrimination (P = 5.8×10-7); adjusting for age, histology, grade, myometrial invasion, TP53 status, and stage, only CCNA2-H/E2F1-H (P = .0003), FBXW7-mu/PPP2R1A-mu (P = .0002), and stage (P = .017) were significant. The generated prognostic molecular classification system identifies dissimilar signaling aberrations potentially amenable to targetable therapeutic options.


Bimodal sedimenting zones due to ligand-mediated interactions.
Ligand-mediated association-dissociation reactions can give rise to band sedimentation patterns showing bimodal bands despite instantaneous establishment of equilibrium. Weaker interactions result in unimodal bands whose sedimentation coefficients decrease with time of sedimentation and in characteristic patterns of total ligand. The implications of these results for fundamental investigations of protein interactions and for conventional analytical applications of zone sedimentation and molecular sieve chromatography are considered.


Slot waveguides with polycrystalline silicon for electrical injection.
We demonstrate horizontal slot waveguides using high-index layers of polycrystalline and single crystalline silicon separated by a 10 nm layer of silicon dioxide. We measure waveguide propagation loss of 7 dB/cm and a ring resonator intrinsic quality factor of 83,000. The electric field of the optical mode is strongly enhanced in the low-index oxide layer, which can be used to induce a strong modal gain when an active material is embedded in the slot. Both high-index layers are made of electrically conductive silicon which can efficiently transport charge to the slot region. The incorporation of conductive silicon materials with high-Q slot waveguide cavities is a key step for realizing electrical tunneling devices such as electrically pumped silicon-based light sources.


The effect of semelil (angipars®) on bone resorption and bone formation markers in type 2 diabetic patients.
Diabetes mellitus has been recognized as a major risk factor for osteoporosis in which bone turnover is affected by different mechanisms. As the morbidity, mortality and financial cost related to osteoporosis are expected to rise in Iran in coming years, and considering the efficacy of Angipars® for improvement of different ulcers which made it a new herbal drug in diabetic foot ulcer, there is a need to evaluate the effect of this new drug on different organs including bone resorption and bone formation markers. In this randomized, double- blind clinical trial, 61 diabetic patients were included. The subjects were randomly divided into intervention and control groups. Subjects of intervention group received 100 mg of Angipars® twice a day. Laboratory tests including bone resorption and bone formation markers were performed at baseline and after 3 months. 31 patients in study group and 30 patients in control group finished the study. The mean age of the study population and the mean disease duration was respectively 51.8 ± 6.2 and 7.5 ± 4.7 years with no significant differences between intervention and control patients. No statistically significant differences between patients and controls were observed in pyridinoline, osteocalcin, urine calcium, bone alkaline phosphatase and tumor necrosis factor (TNF-α). Only urine creatinine level significantly changed between two groups after 3 month of treatment (p-value: 0.029) CONCLUSION: In conclusion, the findings of this study indicate that Semelil (Angipars®) had no beneficial or harmful effects on bone. It might be other effects of this new component on bone turnover process which need more studies and more time to be discovered.


Red ear syndrome - case report and review of literature.
Red ear syndrome is characterized by: paroxysmal, unilateral, recurrent pain, redness and discomfort of the ear lobe accompanied by a burning sensation. The duration and frequency of red ear syndrome attacks is very various and the episodes, usually occur spontaneously. The pathophysiology is still unknown and also there are no medications with approved efficacy. The goal of this brief report is to present a 11-year old girls whose symptoms of red ear syndrome preceded migraine without aura and the signs of redness of the ear occurred in clusters. The occurrence of symptoms of our case may have confirmed the observation that red ear syndrome is associated with primary headaches particularly migraine and cluster headaches. The literature on this case report of pediatric idiopathic red ear syndrome has been reviewed.


Adaptive noise smoothing filter for images with signal-dependent noise.
In this paper, we consider the restoration of images with signal-dependent noise. The filter is noise smoothing and adapts to local changes in image statistics based on a nonstationary mean, nonstationary variance (NMNV) image model. For images degraded by a class of uncorrelated, signal-dependent noise without blur, the adaptive noise smoothing filter becomes a point processor and is similar to Lee's local statistics algorithm [16]. The filter is able to adapt itself to the nonstationary local image statistics in the presence of different types of signal-dependent noise. For multiplicative noise, the adaptive noise smoothing filter is a systematic derivation of Lee's algorithm with some extensions that allow different estimators for the local image variance. The advantage of the derivation is its easy extension to deal with various types of signal-dependent noise. Film-grain and Poisson signal-dependent restoration problems are also considered as examples. All the nonstationary image statistical parameters needed for the filter can be estimated from the noisy image and no a priori information about the original image is required.


Digital Evaluation of Functional Occlusion Parameters and their Association with Temporomandibular Disorders.
Dental researchers are contradictory in their opinion on the role of occlusion in TMD. Occlusal evaluation of both conventional and digital methods in TMD patients will provide the accurate information about the factors accountable for occlusal instability. Identifying the factors responsible will facilitate precise diagnosis and treatment for TMD. The AIM of the study was to determine the dynamic occlusal parameters strongly associated with the etiology of Temporomandibular disorders. Study group consisted of 100 patients; it included 50 patients with normal TMJ(Group I) and remaining 50 patients had a minimum of one positive sign or symptom of Temporomandibular disorder (GroupII). The patient's dynamic occlusal contacts were evaluated by both conventional and digital methods. The Articulating paper was utilized for conventional occlusion analysis. During conventional analysis centric, lateral and protrusive interferences were evaluated along with loss of vertical dimension. Digital occlusal analysis was performed with T-Scan III. Clusion time, disclusion times were recorded for both groups. Chi-square and Student't' statistical analyses were performed to ascertain the association and statistically significant difference between the groups using SPSS19. Group II patients predominantly (66%)had Group-function occlusion compared to Group I subject. Centric slide more than 2 mm found to have strong influence (p value 0.008) on the etiology of TMD. Among the occlusal interferences evaluated balanced side interferences had a strong correlation with TMD with p-value of 0.003. Working side interferences, protrusive interferences had a p-value of 0.157, 0.826 respectively, indicating weak association. T-Scan analysis showed Group I had 0.689, 0.9136, 0.7952, 0.9794 seconds of clusion, left, right, protrusive disclusion time respectively compared to corresponding 1.862, 1.7995, 1.6978, 1.9296 seconds for Group II. Statistically significant difference (p≤0.05) was found between the mean values of both groups. Among the dynamic occlusal parameters evaluated centric slide and balancing side interferences were found to be highly influential in TMD etiology. TMD patients had prolonged clusion and disclusion times compared to healthy TMJ patients.


Outcomes That Matter to Adolescents With Continuous Headache Due to Chronic Migraine and Their Parents: A Pilot Survey Study.
Children and adolescents with chronic migraine who have continuous headache may have high levels of headache-related disability but have largely been excluded from clinical trials. Understanding patient-valued treatment outcomes may facilitate future study design. The aim of this work was to identify patient-valued outcome measures for headache preventive treatments among adolescents with continuous headache due to chronic migraine and their parents. Adolescents with an International Classification of Headache Disorders, 3rd edition diagnosis of chronic migraine, who had continuous headache and who were being cared for at a tertiary pediatric headache clinic, and their parents were surveyed to query the value of treatment outcomes in the domains of headache frequency, intensity, functional impact, and associated symptoms. Individual outcomes and categories of outcomes were ranked in order of preference using weighted average rank. Results were rounded to the degree of precision with which they were measured. The survey was completed by 55 adolescents and 60 parents of adolescents with continuous headache due to chronic migraine. Mean age of adolescents was 16 (SD 1, range 12-17) years. Median adolescent-reported duration of continuous headache was 24 (interquartile range [IQR] 12-39) months, and adolescents had tried a median of 2 (IQR 0-5) preventive medications, only 13% of which were perceived as helpful. Overall, the most valued individual outcome measure among both adolescents and parents was a decrease in frequency of more severe headaches; however, outcomes reflecting headache intensity were most valued by adolescents, while outcomes reflecting functional impact were most valued by parents. More than 60% of adolescents felt that it was more important to measure decrease in frequency and intensity of headaches in terms of severe headaches/spikes rather than total headache days. Among associated symptoms, improvement in brain fog was most highly valued by both adolescents and parents. The results of this study provide important information about which preventive treatment outcomes are valued by adolescents with continuous headache due to chronic migraine and their parents. Results suggest that adolescents value a decrease in frequency of severe headaches over a decrease in frequency of headache days overall. Generalizability may be limited because the surveyed population was being cared for at a tertiary pediatric headache clinic and generally had headache disorders that were refractory to treatment. These results may help guide future trial design in this population with continuous headache due to chronic migraine.


[Serologic prevalence of human leptospirosis in 2 different groups in the province of Formosa, Argentina].
Sera from two different human group from the Province of Formosa were checked for 14 leptospira serovars, using Martin & Petti's microagglutination test with live antigens. In the first group, consisting of 605 males from the whole province, sampled during premilitary medical screening, 20 were found positive (3.3%). The predominant serovars were as follows: L. bratislava (50%), L. pomona (25%), L. butembo and L. icterohemorragica (20%), L. canicola (15%) and last L. wolffii (5%). In the second group, the sera were obtained from the local Central Hospital, and were taken from adults of both sexes, from a limited area of the province. With regard to Group I, there was a high number of positive (32.3%) and a variation in the frequency of serovars, the results being as follows: L. butembo (38.8%), L. pomona (33.3%), L. bratislava (31.4%), L. icterohemorragica (12.9%), L. borincana (3.7%) and finally L. canicola, L. pyrogenes, L. wolffii and L. bataviae (1.8%).


Cross-reactive sensor arrays for the detection of peptides in aqueous solution by fluorescence spectroscopy.
A simple but powerful method for the sensing of peptides in aqueous solution has been developed. The transition-metal complexes [PdCl(2)(en)], [{RhCl(2)Cp*}(2)], and [{RuCl(2)(p-cymene)}(2)] were combined with six different fluorescent dyes to build a cross-reactive sensor array. The fluorescence response of the individual sensor units was based on competitive complexation reactions between the peptide analytes and the fluorescent dyes. The collective response of the sensor array in a time-resolved fashion was used as an input for multivariate analyses. A sensor array comprised of only six metal-dye combinations was able to differentiate ten different dipeptides in buffered aqueous solution at a concentration of 50 muM. Furthermore, the cross-reactive sensor could be used to obtain information about the identity and the quantity of the pharmacologically interesting dipeptides carnosine and homocarnosine in a complex biological matrix, such as deproteinized human blood serum. The sensor array was also able to sense longer peptides, which was demonstrated by differentiating mixtures of the nonapeptide bradykinin and the decapeptide kallidin.


Clinical and cognitive insight in patients with acute-phase psychosis: Association with treatment and neuropsychological functioning.
The severity of psychopathology cannot fully explain deficits in the multi-dimensional construct of insight. The aim of this study was to evaluate the correlates and associations of clinical and cognitive insight in patients in an acute phase of psychosis and to analyse the impact of acute treatment on these variables. This study examined 47 inpatients who were recently hospitalized with acute exacerbation of schizophrenia. All subjects were assessed at both admission and discharge with the Positive and Negative Syndrome Scale (PANSS), Schedule for the Assessment of Insight-Expanded Version (SAI-E), Beck Cognitive Insight Scale (BCIS), and a neurocognition battery. Patients with schizophrenia gained clinical insight after treatment. Cognitive insight did not change significantly after treatment. Insight showed significant negative correlations with positive symptoms and general psychopathology, but not with negative symptoms. Clinical insight was not associated with neuropsychological functioning in this cohort. Gaining clinical insight in the acute phase of illness was associated with the remission of positive symptoms, but not with neuropsychological functioning. Some significant correlations between clinical and cognitive insights were detected, which suggests that cognitive insight contributes to clinical insight but is not treatment-dependent. Long-term treatment may be required to understand the contribution of insight to the outcome of patients with schizophrenia.


Using iron oxide nanoparticles to diagnose CNS inflammatory diseases and PCNSL.
The study goal was to assess the benefits and potential limitations in the use of ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles in the MRI diagnosis of CNS inflammatory diseases and primary CNS lymphoma. Twenty patients with presumptive or known CNS lesions underwent MRI study. Eighteen patients received both gadolinium-based contrast agents (GBCAs) and 1 of 2 USPIO contrast agents (ferumoxytol and ferumoxtran-10) 24 hours apart, which allowed direct comparative analysis. The remaining 2 patients had only USPIO-enhanced MRI because of a renal contraindication to GBCA. Conventional T1- and T2-weighted MRI were acquired before and after contrast administration in all patients, and perfusion MRI for relative cerebral blood volume (rCBV) assessment was obtained in all 9 patients receiving ferumoxytol. USPIO-enhanced MRI showed an equal number of enhancing brain lesions in 9 of 18 patients (50%), more enhancing lesions in 2 of 18 patients (11%), and fewer enhancing lesions in 3 of 18 patients (17%) compared with GBCA-enhanced MRI. Four of 18 patients (22%) showed no MRI enhancement. Dynamic susceptibility-weighted contrast-enhanced perfusion MRI using ferumoxytol showed low rCBV (ratio <1.0) in 3 cases of demyelination or inflammation, modestly elevated rCBV in 5 cases of CNS lymphoma or lymphoproliferative disorder (range: 1.3-4.1), and no measurable disease in one case. This study showed that USPIO-enhanced brain MRI can be useful in the diagnosis of CNS inflammatory disorders and lymphoma, and is also useful for patients with renal compromise at risk of nephrogenic systemic fibrosis who are unable to receive GBCA.


Statin use is associated with improved survival in multiple myeloma: A Swedish population-based study of 4315 patients.
Statin use has been associated with reduced cancer-specific mortality among patients with several cancer types, including multiple myeloma (MM). We aimed to further elucidate the association of statin use and dose intensity with MM survival. Using Swedish population-based national health registers, we identified all incident MM diagnoses occurring January 1, 2007 to December 31, 2013 and their drug dispensations and comorbidities. We assessed statin exposure in 6-month periods pre- and post-diagnosis, treated diagnosis as baseline for calculating survival time, and calculated hazard ratios (HR) and 95% confidence intervals (CI) of exposure-related MM-specific and all-cause mortality using Cox regression. We assessed statin exposure during the entire follow-up and risk of MM-specific mortality in a nested case-control analysis. We classified dose intensity according to American College of Cardiology/American Heart Association recommendations. We ascertained 4315 MM cases during follow-up. Statin use was associated with reduced MM-specific mortality (pre-diagnosis use multivariate-adjusted HR, 95% CI: 0.83, 0.71-0.96; 6 months post-diagnosis: 0.73, 0.60-0.89; entire follow-up: 0.65, 0.52-0.80) and (more weakly) with all-cause mortality. Intensity analyses suggested a dose-response; MM-specific mortality decreased with increasing statin intensity in all time windows (eg, 6 months post-diagnosis: low [0.76 (0.56-1.03)], medium [0.73 (0.58-0.92)], high [0.33 (0.08-1.32)] intensity). However, relatively few patients received high intensity treatment, and the trend was statistically significant only for unadjusted pre-diagnosis use. In this large population-based MM cohort, statin use was associated with improved MM-specific survival in both sexes. Randomized prospective studies are warranted to evaluate statins as adjuvant treatment in MM.


Convergent Validity and Preliminary Cut-Off Scores for the Anxiety and Depression Subscales of the DASS-21 in US Adolescents.
Although the depression and anxiety subscales (DASS-D and DASS-A) of the 21-item Depression, Anxiety, and Stress Scales are commonly used in adolescents, few studies have investigated the convergent validity of/potential cutoff scores for these subscales in U.S. adolescents. To address these gaps, 306 U.S. adolescents completed the DASS-21, Generalized Anxiety Disorder-7 scale (GAD7), and Center for Epidemiological Studies-Depression Scale (CESD). The DASS-D and CESD were strongly correlated, while the DASS-A and GAD7 were strongly correlated in males and more moderately correlated in females. Using the CESD as reference, DASS-D cutoff scores of four for males and five for females provided the best balance of sensitivity and specificity. When using the GAD7 as reference, DASS-A cutoffs of six for females and five for males appeared best. These findings suggest the DASS-A and DASS-D demonstrate good convergent validity and may be suitable for identifying adolescents who are significantly anxious and/or depressed.


Evaluation of adherence to therapy in patients of chronic kidney disease.
To evaluate adherence to medication and study factors associated with non-adherence in chronic kidney disease (CKD) patients. A prospective, cross-sectional, questionnaire based study was conducted in Nephrology department of a super specialty hospital. Patients above 18 years of age, suffering from CKD from six months or more were interviewed using self-designed, semi-structured questionnaire to get information about adherence to medication, diet restriction and lifestyle modification (n = 150). Morisky medication adherence questionnaire was used to calculate overall adherence. In this higher score indicates poor adherence. Main outcome measures included prevalence of non-adherence and factors associated with the same. Average number of medicines taken by each patient was 8.0+1.612 (mean+SD) per day. Non-adherence to medication schedule was reported in 34% patients. Common causes of non-adherence were high cost (21.3%), complex dosing schedule (20%), fear of adverse effects (16%). Sixty-eight% patients were not aware about importance of taking each medicine. Sixteen% stopped taking medicines due to high cost. Forty-two% suggested that government should adopt measures to provide free medicines to poor patients. In Morisky medication adherence questionnaire high, medium and low adherence was reported in 7.3%, 55.3% and 37.3% of patients, respectively. Moderately positive correlation was observed between poor adherence and number of concurrent illnesses and number of medicines taken. Since majority of patients were not aware about importance of taking each medicine, creating awareness about the same is essential for improving adherence to therapy. Measures to provide free medicines to non-affording patients need to be implemented since high cost was other major cause of non-adherence.


Ethical and Legal Obligations for Research Involving Pregnant Persons in a Post-Dobbs Context.
In light of a history of categorical exclusion, it is critical that pregnant people are included in research to help improve the knowledge base and interventions needed to address public health. Yet the volatile legal landscape around reproductive rights in the United States threatens to undue recent progress made toward the greater inclusion of pregnant people in research. We offer ethical and practical guidance for researchers, sponsors, and institutional review boards to take specific steps to minimize legal risks and ensure the ethical conduct of research with pregnant people in an evolving legal environment.


Clinical Significance of Fractional Anisotropy Measured in Peritumoral Edema as a Biomarker of Overall Survival in Glioblastoma: Evidence Using Correspondence Analysis.
Fractional anisotropy (FA), a diffusion tensor image (DTI) derived biomarker is related to invasion, infiltration, and extension of glioblastoma (GB). We aimed to evaluate FA values and their association with intervals of overall survival (OS). Retrospective study conducted in 36 patients with GB included 23 (63.9%) males, 46 ± 14 y; and 13 (36.1%) females, 53 ± 13; followed up for 36 months. We measured FA at edema, enhancing rim, and necrosis. We created two categorical variables using levels of FA and intervals of OS to evaluate their relationships. Kaplan-Meier method and correspondence analysis evaluated the association between OS (grouped in 7 six-month intervals) and FA measurements. Median FA values were higher in healthy brain regions (0.351), followed by peritumoral edema (0.190), enhancing ring (0.116), and necrosis (0.071). Pair-wise comparisons among tumor regions showed a significant difference, P < 0.001. The median OS for all patients was 19.3 months; variations in the OS curves among subgroups was significant χ2 (3) = 8.48, P = 0.037. Correspondence analysis showed a significant association between FA values in the edema region and the survival intervals χ2 (18) = 30.996, P = 0.029. Alternative multivariate assessment using correspondence analysis might supplement the traditional survival analysis in patients with GB. A close follow-up of the variability of FA in the peritumoral edema region is predictive of the OS within specific six-month interval subgroup. Further studies should focus on predictive models combining surgical and DTI biomarkers.


Long-term pattern of opioid prescriptions after femoral shaft fractures.
The use of opioids in non-cancer-related pain following skeletal trauma is controversial due to the presumed risk of dose escalation and dependence. We therefore examined the pattern of opioid prescriptions, that is, those actually dispensed, in patients with femoral shaft fractures. We analysed data from the Swedish National Hospital Discharge Register and the Swedish Prescribed Drug Register between 2005 and 2008. We identified 1471 patients with isolated femoral shaft fractures. The median age was 75 (16-102) years and 56% were female. In this cohort, 891 patients (61%) received dispensed opioid prescriptions during a median follow-up of 20 months (interquartile range 11-32). In the age- and sex-matched comparison cohort (7339 individuals) without fracture, 25% had opioid prescriptions dispensed during the same period. The proportions of patients receiving opioid analgesics at 6 and 12 months after the fracture were 45% (95% CI 42-49) and 36% (32-39), respectively. The median daily morphine equivalent dose (MED) was between 15 and 17 mg 1-12 months post-fracture. After 3 months, less than 5% used prescription doses higher than 20 mg MED per day. Older age (≥ 70 compared with < 70 years) was a significant predictor of earlier discontinuation of opioid use (Hazard ratio [HR] 1.9). A notable proportion of patients continued to receive dispensed prescriptions for opioids for over 6 months (45%) and more than a third of them (36%) continued treatment for at least 12 months. However, the risk of dose escalation seems to be small in opioid-naïve patients.


Retrograde axonal transport of BDNF and proNGF diminishes with age in basal forebrain cholinergic neurons.
Basal forebrain cholinergic neurons (BFCNs) are critical for learning and memory and degenerate early in Alzheimer's disease (AD). BFCNs depend for their survival and function on nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), which are retrogradely transported from BFCN targets. Age is the greatest risk factor for developing AD, yet the influence of age on BFCN axonal transport is poorly understood. To model aging, embryonic rat basal forebrain or cortical neurons were cultured in microfluidic chambers. Senescence-associated beta-galactosidase staining indicated an aging phenotype only in BFCNs cultured for 18+ days in vitro. BDNF axonal transport impairments were observed exclusivley in aged BFCNs. BFCNs displayed robust proNGF transport, which also diminished with in vitro age. The expression of NGF receptor tropomyosin-related kinase-A and BDNF receptor tropomyosin-related kinase-B also decreased significantly with in vitro age in BFCNs only. These results suggest a unique vulnerability of BFCNs to age-induced transport deficits. These deficits, coupled with the reliance of BFCNs on neurotrophin transport, may explain their vulnerability to age-related neurodegenerative disorders like AD.


Combination therapy with pulsed methylprednisolone in rheumatoid arthritis.
Pulsed methylprednisolone (PMP) has been shown to produce clinical improvement and reduction in the ESR and acute phase protein concentrations in patients with active rheumatoid arthritis and has been advocated for use either as an alternative to slow-acting antirheumatoid drugs (SAARDs) or in conjunction with SAARDs to accelerate the response to treatment. To test these potential roles for PMP 45 patients with active RA were randomly allocated to treatment with PMP alone, PMP + sulphasalazine (SAS - at a maintenance dose of 2.0 g/day), or PMP + D-penicillamine (DPA - at a maintenance dose of 500 mg/day). In each case three 1 g intravenous infusions were given on alternate days during the first week of the trial. Patients were monitored for 24 weeks by standard clinical and laboratory measurements. All three treatment groups showed significant clinical and laboratory improvements at two weeks. With PMP + DPA and PMP + SAS these improvements were sustained and were not significantly different in these two treatment groups. However, in the 'PMP only' group ESR and CRP rose to pretreatment values by eight weeks. Twelve patients withdrew from the study owing to a relapse of the RA. No serious adverse effects were seen in the 'PMP only' group. Both combination regimens were well tolerated; adverse effects seen were attributable to either DPA or SAS. We conclude that PMP alone is insufficient for treatment of RA but can be used successfully in combination with either DPA or SAS. A comparison between these results obtained from two previous groups of 15 patients treated with DPA alone and SAS alone (using the same study design) shows that PMP accelerated the response to therapy by at least six weeks.


HIV type 1 glycoprotein 120 amplifies tumor necrosis factor-induced NF-kappa B activation in Jurkat cells.
This article demonstrates that human immunodeficiency virus type 1 (HIV-1) gp120 amplifies the activity of tumor necrosis factor alpha (TNF-alpha), a cytokine that stimulates HIV-1 replication through activation of NF-kappa B. In CD4-positive Jurkat cells, gp120 potentiates TNF-induced NF-kappa B activation. TNF-mediated activation of NF-kappa B is known to involve the intracellular formation of reactive oxygen intermediates (ROIs). Accordingly, we examined the influence of gp120 on the cellular redox state. We found that gp 120-modulated TNF-induced NK-kappa B activation was inhibited by the antioxidant butylated hydroxyanisole, indicating the involvement of redox-dependent mechanisms. In addition, we showed that gp120 induces intracellular formation of hydrogen peroxide, which is accompanied by a decrease in the ratio of glutathione to glutathione disulfide. In contrast, in the p56lck-deficient J.CaM1.6 T cell line, a derivative of the Jurkat cell line, gp120 was unable to stimulate hydrogen peroxide, to decrease the ratio of GSH to GSSG, and has no effect on TNF-induced NF-kappa B activation. This demonstrated that p56lck protein tyrosine kinase plays an active role in transmitting a signal that increases the oxidative state of the cell and as a consequence amplifies TNF-mediated NF-kappa B DNA binding. We have demonstrated that Tat protein decreased both the Mn-dependent superoxide dismutase (MnSOD) and the cellular glutathione content (GSH). Here we show that, in contrast to Tat, gp120 is unable to inhibit activity and expression of MnSOD and to decrease GSH content. Taken together, our data suggest that gp120 potentiates TNF-induced NF-kappa B activation by stimulating a signal pathway that involves p56lck and the increased formation of reactive oxygen intermediates such as H2O2. These findings may be relevant for the regulation of HIV-1 replication in T cells.


Effect of a multidisciplinary amyotrophic lateral sclerosis (ALS) clinic on ALS survival: a population based study, 1996-2000.
In recent years, there has been a paradigm shift in the method of healthcare delivery to amyotrophic lateral sclerosis (ALS) patients with the emergence of multidisciplinary ALS clinics that cater exclusively for patients with this condition. The impact of multidisciplinary management has not been previously evaluated. Using data from the Irish ALS Register, we conducted a prospective, population based study of all ALS cases diagnosed in Ireland over a five year period to evaluate the effectiveness of a multidisciplinary clinic on ALS survival. Eighty two (24%) patients attended the multidisciplinary ALS clinic, with the remaining 262 (76%) cases followed in a general neurology clinic. The ALS clinic cohort was an average of five years younger (60.1 v 65.6 years) and were more likely to receive riluzole than the general neurology cohort (99% v 61%). The median survival of the ALS clinic cohort was 7.5 months longer than for patients in the general neurology cohort (logrank = 15.4, p < 0.0001). Overall, one year mortality was decreased by 29.7%. Prognosis of bulbar onset patients was extended by 9.6 months if they attended the ALS clinic. Using multivariate analysis, attendance at the ALS clinic was an independent covariate of survival (HR = 1.47, p = 0.02). ALS patients who received their care at a multidisciplinary clinic had a better prognosis than patients attending a general neurology clinic. The data suggest that active and aggressive management enhances survival, particularly among ALS patients with bulbar dysfunction. The effect of clinic type must be considered in future clinical trials design.


Apigenin inhibits TNFα/IL-1α-induced CCL2 release through IKBK-epsilon signaling in MDA-MB-231 human breast cancer cells.
Mortality associated with breast cancer is attributable to aggressive metastasis, to which TNFα plays a central orchestrating role. TNFα acts on breast tumor TNF receptors evoking the release of chemotactic proteins (e.g. MCP-1/CCL2). These proteins direct inward infiltration/migration of tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), myeloid-derived suppressor cells (MDSCs), T-regulatory cells (Tregs), T helper IL-17-producing cells (Th17s), metastasis-associated macrophages (MAMs) and cancer-associated fibroblasts (CAFs). Tumor embedded infiltrates collectively enable immune evasion, tumor growth, angiogenesis, and metastasis. In the current study, we investigate the potential of apigenin, a known anti-inflammatory constituent of parsley, to downregulate TNFα mediated release of chemokines from human triple-negative cells (MDA-MB-231 cells). The results show that TNFα stimulation leads to large rise of CCL2, granulocyte macrophage colony-stimulating factor (GMCSF), IL-1α and IL-6, all suppressed by apigenin. While many aspects of the transcriptome for NFkB signaling were evaluated, the data show signaling patterns associated with CCL2 were blocked by apigenin and mediated through suppressed mRNA and protein synthesis of IKBKe. Moreover, the data show that the attenuation of CCL2 by apigenin in the presence TNFα paralleled the suppression of phosphorylated extracellular signal-regulated kinase 1 (ERK 1/ 2). In summary, the obtained findings suggest that there exists a TNFα evoked release of CCL2 and other LSP recruiting cytokines from human breast cancer cells, which can be attenuated by apigenin.


Regulation of tyrosinase in human melanocytes grown in culture.
Tyrosinase, the enzyme that controls the synthesis of melanin, is a unique product of melanocytes. Normal and malignant human melanocytes grown in culture were used to study the factors that regulate the expression of tyrosinase. Immunoprecipitation experiments showed that newly synthesized tyrosinase appeared as a protein with an apparent molecular weight of 70,000 that was processed to a protein with an apparent molecular weight of 80,000. Neither tunicamycin nor 2-deoxy-D-glucose inhibited this conversion, suggesting that O-glycosylation is the major biochemical event in the posttranslational modification of tyrosinase. Agents that stimulated the proliferation of normal melanocytes also stimulated tyrosinase activity. Melanocytes with low levels of tyrosinase activity synthesized less tyrosinase, processed the enzyme more slowly, and degraded it more rapidly than melanocytes with high levels of tyrosinase activity. We conclude that tyrosinase activity in cultures of human melanocytes derived from different donors is determined predominantly by its abundance.


Divergent unsaturated fatty acid synthesis in two highly related model pseudomonads.
The genomes of the best-studied pseudomonads, Pseudomonas aeruginosa and Pseudomonas putida, which share 85% of the predicted coding regions, contain a fabA fabB operon (demonstrated in P. aeruginosa, putative in P. putida). The enzymes encoded by the fabA and fabB genes catalyze the introduction of a double bond into a 10-carbon precursor which is elongated to the 16:1Δ9 and 18:1Δ11 unsaturated fatty acyl chains required for functional membrane phospholipids. A detailed analysis of transcription of the P. putida fabA fabB gene cluster showed that fabA and fabB constitute an operon and disclosed an unexpected and essential fabB promoter located within the fabA coding sequence. Inactivation of the fabA fabB operon fails to halt the growth of P. aeruginosa PAO1 but blocks growth of P. putida F1 unless an exogenous unsaturated fatty acid is provided. We report that the asymmetry between these two species is due to the P. aeruginosa PAO1 desA gene which encodes a fatty acid desaturase that introduces double bonds into the 16-carbon acyl chains of membrane phospholipids. Although P. putida F1 encodes a putative DesA homolog that is 84% identical to the P. aeruginosa PAO1, the protein fails to provide sufficient unsaturated fatty acid synthesis for growth when the FabA FabB pathway is inactivated. We report that the P. putida F1 DesA homolog can functionally replace the P. aeruginosa DesA. Hence, the defect in P. putida F1 desaturation is not due to a defective P. putida F1 DesA protein but probably to a weakly active component of the electron transfer process.


IL28B polymorphism and the control of hepatitis C virus infection: ready for clinical use?
Polymorphisms in the region of the interleukin-28B (IL28B) gene have recently been associated with spontaneous and treatment induced clearance of hepatitis C virus infection. The specific mechanisms of how IL28B polymorphisms affect HCV suppression remain unknown. It is a matter of ongoing debate how to incorporate the IL28B data into the current treatment algorithms with pegylated interferon-alpha and ribavirin. The eventual role of the IL28B genotype in new therapeutic regimes with direct antiviral agents needs to be explored in the ongoing and future clinical studies with these agents.


[Breast-feeding training programme as intervention approach at the hospital level: results of evaluation of the first phase of the STELLA study].
The health-promoting short- and long-term effects of breast-feeding are supported by a vast scientific literature. The Bavarian prospective cohort study 2005/2006 showed regional variations of breast-feeding rates. Furthermore, improvement in counselling mothers, particularly with regard to prevention and handling of breast-feeding problems, has a great potential for health promotion. The objective of this study is to promote breast-feeding in maternity clinics by improving the surrounding conditions. A clinic-based and community-related intervention trial was carried out in Lower Bavaria as a model region with below average breast-feeding rates. (1) INTERVENTION: an advanced training of maternity ward professionals of 10 hospitals and after-care midwives was performed from May until December 2008. The training programme was based on the WHO/UNICEF criteria of the "Ten Steps to Successful Breastfeeding" to deepen the breast-feeding knowledge and to improve the breast-feeding management. (2) EVALUATION: A singular assessment of the advanced training programme was undertaken concerning an increase of knowledge and the practicability in clinical everyday life by participants; improvements of the maternity wards were assessed via structural interviews with maternity ward staff before and after intervention. Approximately 85% (n=378) of the staff of the 10 maternity clinics attended the training course. The survey after the training programme (response rate 83%) indicated that more than 80% of the participants stated to have learned something new and to be able to use the knowledge acquired for their own practice. Results of the clinic interviews showed a transfer of training contents into clinical work routines. Improvements at interview date were shown best for bonding (all 10 maternity clinics), for additional feeding (8 maternity clinics) as well as for 24-h rooming-in and mother counselling (7 maternal clinics each). Training of maternity ward professionals accounts for baby-friendly conditions in maternity clinics. Most willingness for improvements was shown by maternity ward staff particularly for bonding between mother and child as well as for additional feeding. Considerable differences between the hospitals were observed for the promotion of infant formulas before and after the intervention.


Involvement of insulin-like growth factor-I and insulin-like growth factor binding proteins in pro-B-cell development.
Insulin-like growth factor (IGF)-binding proteins (IGFBPs) are a family of proteins thought to modulate IGF function. By employing an in vitro culture system of human hematopoietic stem cells cocultured with murine bone marrow stromal cells, we examined the effects of IGF-I and IGFBPs on early B-cell development. Human CD34(+) bone marrow cells were cocultured with murine stromal MS-5 cells for 4 weeks, and pro-B-cell number was analyzed by flow cytometry. After administration of reagents that are supposed to modulate IGF-I or IGFBP function to the culture, the effect on pro-B-cell development was examined. After cultivation for 4 weeks, effective induction of pro-B-cell proliferation was observed. Experiments using several distinct factors, all of which neutralize IGF-I function, revealed that impairment of IGF-I function results in a significant reduction in pro-B-cell development from CD34(+) cells. In addition, when the effect of recombinant proteins of IGFBPs and antibodies against IGFBPs were tested, IGFBP-3 was found to inhibit pro-B-cell development, while IGFBP-6 was required for pro-B-cell development. IGF-I is essential for development of bone marrow CD34(+) cells into pro-B cells. Moreover, IGFBPs are likely involved in regulation of pro-B-cell development.


Role of ferulic acid in the amelioration of ionizing radiation induced inflammation: a murine model.
Ionizing radiation is responsible for oxidative stress by generating reactive oxygen species (ROS), which alters the cellular redox potential. This change activates several redox sensitive enzymes which are crucial in activating signaling pathways at molecular level and can lead to oxidative stress induced inflammation. Therefore, the present study was intended to assess the anti-inflammatory role of ferulic acid (FA), a plant flavonoid, against radiation-induced oxidative stress with a novel mechanistic viewpoint. FA was administered (50 mg/kg body wt) to Swiss albino mice for five consecutive days prior to exposing them to a single dose of 10 Gy 60Co γ-irradiation. The dose of FA was optimized from the survival experiment and 50 mg/kg body wt dose showed optimum effect. FA significantly ameliorated the radiation induced inflammatory response such as phosphorylation of IKKα/β and IκBα and consequent nuclear translocation of nuclear factor kappa B (NF-κB). FA also prevented the increase of cycloxygenase-2 (Cox-2) protein, inducible nitric oxide synthase-2 (iNOS-2) gene expression, lipid peroxidation in liver and the increase of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in serum. It was observed that exposure to radiation results in decreased activity of superoxide dismutase (SOD), catalase (CAT) and the pool of reduced glutathione (GSH) content. However, FA treatment prior to irradiation increased the activities of the same endogenous antioxidants. Thus, pretreatment with FA offers protection against gamma radiation induced inflammation.


Modeling Schistosoma japonicum Infection under Pure Specification Bias: Impact of Environmental Drivers of Infection.
Uncertainties in spatial modeling studies of schistosomiasis (SCH) are relevant for the reliable identification of at-risk populations. Ecological fallacy occurs when ecological or group-level analyses, such as spatial aggregations at a specific administrative level, are carried out for an individual-level inference. This could lead to the unreliable identification of at-risk populations, and consequently to fallacies in the drugs' allocation strategies and their cost-effectiveness. A specific form of ecological fallacy is pure specification bias. The present research aims to quantify its effect on the parameter estimates of various environmental covariates used as drivers for SCH infection. This is done by (i) using a spatial convolution model that removes pure specification bias, (ii) estimating group and individual-level covariate regression parameters, and (iii) quantifying the difference between the parameter estimates and the predicted disease outcomes from the convolution and ecological models. We modeled the prevalence of Schistosoma japonicum using group-level health outcome data, and city-level environmental data as a proxy for individual-level exposure. We included environmental data such as water and vegetation indexes, distance to water bodies, day and night land surface temperature, and elevation. We estimated and compared the convolution and ecological model parameter estimates using Bayesian statistics. Covariate parameter estimates from the convolution and ecological models differed between 0.03 for the nearest distance to water bodies (NDWB), and 0.28 for the normalized difference water index (NDWI). The convolution model presented lower uncertainties in most of the parameter estimates, except for NDWB. High differences in uncertainty were found in night land surface temperature (0.23) and elevation (0.13). No significant differences were found between the predicted values and their uncertainties from both models. The proposed convolution model is able to correct for a pure specification bias by presenting less uncertain parameter estimates. It shows a good predictive performance for the mean prevalence values and for a positive number of infected people. Further research is needed to better understand the spatial extent and support of analysis to reliably explore the role of environmental variables.


Cerebrospinal fluid lymphocyte subpopulations before and after pneumoencephalography.
Pneumoencephalography (PEG) did not cause significant changes in cerebrospinal fluid (CSF) lymphocyte subpopulations. Sheep erythrocyte rosette-forming cells (E-RFC) percentage was higher in CSF before (84.0) and after (81.1) PEG than in blood (63.7). The percentage of complement receptor bearing cells was significantly higher in CSF (65.3 and 64.6) than in blood (33.1). Fcgamma receptor bearing cell levels were only slightly higher in CSF (35.6%) than in blood (31.5%), and increased after PEG to 43.9%. The high percentages of both E-RFC and complement receptor bearing cells in CSF, suggest that a considerable number of these cells bear both receptors, in contrast to blood.


Development and validation of a UPLC-MS/MS method for determination of SYHA1807 in a first-in-human study.
Background: SYHA1807 is a novel lysine specific demethylase 1 inhibitor being developed for the treatment of small-cell lung cancer. Aim: This study aimed to establish a ultra-performance liquid chromatography-mass spectrometry (UPLC-MS)/MS method for measuring SYHA1807 in human plasma, supporting its application in a first-in-human study. Methods: SYHA1807 was separated on an ACQUITY UPLC BEH® C18 Column (2.1 × 50 mm, 1.7 μm) after protein precipitation of plasma samples. Mass spectrometry analysis was performed with a Xevo TQS triple quadrupole mass spectrometer utilizing a positive electronic spray ionization source. The established method was fully validated according to bioanalytical guidelines. Results & conclusion: A rapid, specific and robust UPLC-MS/MS method was first established for quantifying SYHA1807 and successfully applied in a first-in-human study.


Pilot study on the urinary excretion of the glyphosate metabolite aminomethylphosphonic acid and breast cancer risk: The Multiethnic Cohort study.
Breast cancer is the most commonly diagnosed female cancer and the second leading cause of death in women in the US, including Hawaii. Accumulating evidence suggests that aminomethylphosphonic acid (AMPA), the primary metabolite of the herbicide glyphosate-a probable human carcinogen, may itself be carcinogenic. However, the relationship between urinary AMPA excretion and breast cancer risk in women is unknown. In this pilot study, we investigated the association between pre-diagnostic urinary AMPA excretion and breast cancer risk in a case-control study of 250 predominantly postmenopausal women: 124 cases and 126 healthy controls (individually matched on age, race/ethnicity, urine type, date of urine collection, and fasting status) nested within the Hawaii biospecimen subcohort of the Multiethnic Cohort. AMPA was detected in 90% of cases and 84% of controls. The geometric mean of urinary AMPA excretion was nearly 38% higher among cases vs. controls (0.087 vs 0.063 ng AMPA/mg creatinine) after adjusting for race/ethnicity, age and BMI. A 4.5-fold higher risk of developing breast cancer in the highest vs. lowest quintile of AMPA excretion was observed (ORQ5 vs. Q1: 4.49; 95% CI: 1.46-13.77; ptrend = 0.029). To our knowledge, this is the first study to prospectively examine associations between urinary AMPA excretion and breast cancer risk. Our preliminary findings suggest that AMPA exposure may be associated with increased breast cancer risk; however, these results require confirmation in a larger population to increase study power and permit careful examinations of race/ethnicity differences.


Medicare cost limits ... is there any relief?
Medicare cost limits stem from 20-year-old legislation that determines "reasonable" rates for reimbursement. Although those rates have periodically been adjusted, they often do not represent a reasonable limitation on a provider's reimbursement. How can agencies challenge limits and appeal unfavorable rulings? What are the most successful means of appeal?


A Latex Metabolite Benefits Plant Fitness under Root Herbivore Attack.
Plants produce large amounts of secondary metabolites in their shoots and roots and store them in specialized secretory structures. Although secondary metabolites and their secretory structures are commonly assumed to have a defensive function, evidence that they benefit plant fitness under herbivore attack is scarce, especially below ground. Here, we tested whether latex secondary metabolites produced by the common dandelion (Taraxacum officinale agg.) decrease the performance of its major native insect root herbivore, the larvae of the common cockchafer (Melolontha melolontha), and benefit plant vegetative and reproductive fitness under M. melolontha attack. Across 17 T. officinale genotypes screened by gas and liquid chromatography, latex concentrations of the sesquiterpene lactone taraxinic acid β-D-glucopyranosyl ester (TA-G) were negatively associated with M. melolontha larval growth. Adding purified TA-G to artificial diet at ecologically relevant concentrations reduced larval feeding. Silencing the germacrene A synthase ToGAS1, an enzyme that was identified to catalyze the first committed step of TA-G biosynthesis, resulted in a 90% reduction of TA-G levels and a pronounced increase in M. melolontha feeding. Transgenic, TA-G-deficient lines were preferred by M. melolontha and suffered three times more root biomass reduction than control lines. In a common garden experiment involving over 2,000 T. officinale individuals belonging to 17 different genotypes, high TA-G concentrations were associated with the maintenance of high vegetative and reproductive fitness under M. melolontha attack. Taken together, our study demonstrates that a latex secondary metabolite benefits plants under herbivore attack, a result that provides a mechanistic framework for root herbivore driven natural selection and evolution of plant defenses below ground.


Role of snail activation in alcohol-induced iNOS-mediated disruption of intestinal epithelial cell permeability.
Chronic alcohol use results in many pathological effects including alcoholic liver disease (ALD). ALD pathogenesis requires endotoxemia. Our previous studies showed that increased intestinal permeability is the major cause of endotoxemia, and that this gut leakiness is dependent on alcohol stimulation of inducible nitric oxide synthase (iNOS) in both alcoholic subjects and rodent models of alcoholic steatohepatitis. The mechanism of the alcohol-induced, iNOS-mediated disruption of the intestinal barrier function is not known. We have recently shown that alcohol stimulates activation of the transcription factor Snail and biomarkers of epithelial mesenchymal transition. As activated Snail disrupts tight junctional proteins, we hypothesized that activation of Snail by iNOS might be one of the key signaling pathways mediating alcohol-stimulated intestinal epithelial cell hyperpermeability. We measured intestinal permeability in alcohol-fed C57BL/6 control and iNOS knockout (KO) mice, and measured Snail protein expression in the intestines of these mice. We then examined intestinal epithelial permeability using the Caco-2 cell model of the intestinal barrier ± small interfering RNA (siRNA) inhibition of Snail. We assessed Snail activation by alcohol in Caco-2 cells ± inhibition of iNOS with L-NIL or siRNA. Finally, we assessed Snail activation by alcohol ± inhibition with siRNA for p21-activated kinase (PAK1). Our data show that chronic alcohol feeding promotes intestinal hyperpermeability in wild-type BL/6, but not in iNOS KO mice. Snail protein expression was increased in the intestines of alcohol-treated wild-type mice, but not in iNOS KO mice. siRNA inhibition of Snail significantly inhibited alcohol-induced hyperpermeability in Caco-2 cell monolayers. Alcohol stimulation of Snail(pS246) activation was blocked by inhibition of iNOS with L-NIL or with siRNA. siRNA inhibition of PAK1 significantly inhibited alcohol-mediated activation of Snail in Caco-2 cells. Our data confirmed our prior results and further demonstrated that alcohol-induced gut leakiness in rodents and intestinal epithelial cell monolayers is iNOS dependent. Our data also support a novel role for Snail activation in alcohol-induced, iNOS-mediated intestinal hyperpermeability and that PAK1 is responsible for activation of Snail at Ser246 with alcohol stimulation. Identification of these mechanisms for alcohol-induced intestinal hyperpermeability may provide new therapeutic targets for prevention and treatment of alcohol-induced leaky gut, endotoxemia, and endotoxin-associated complications of alcoholism such as ALD.


Twenty years of molecular analyses in amyotrophic lateral sclerosis: genetic landscape of Italian patients.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with a heterogeneous genetic background. Because mutation analysis by Sanger sequencing is costly and time-consuming, in recent years, next-generation sequencing (NGS) techniques have become of much interest. This study analyses the results of 20 years of molecular analyses in ALS patients in our laboratory using traditional methods and NGS. Almost 300 ALS patients underwent genetic analysis with Sanger sequencing of 7 genes or with an NGS panel of 23 genes. The C9orf72 expansion was tested by fragment size analysis. Sanger sequencing revealed mutations in 23.8% of familial and 3.8% of sporadic cases, whereas NGS detected potentially pathogenic variants in 45.5% of familial and 5.4% of sporadic cases and variants of unknown significance in 30.3% of patients. In 11.8% of patients, potentially causative mutations were found in 2 or more ALS genes. Compared to traditional methods, NGS is more effective in revealing possibly causal variants, but counseling patients becomes more complicated due to frequent variants of unknown significance and potentially oligogenic cases.


Mechanisms of the inflammatory response.
The physiological alterations induced by acute inflammation present significant management challenges for anaesthesiologists. Major surgery, trauma, burns and sepsis all have large inflammatory components. Acute inflammation is characterized by vasodilatation, fluid exudation and neutrophil infiltration. These processes are activated and amplified by a series of intracellular and extracellular factors that tightly co-ordinate the inflammatory process. The innate immune system responds rapidly to infection or injury. Macrophages, natural killer cells, CD8 + T-lymphocytes and neutrophils provide an early response to injurious factors in an effort to contain and eliminate harmful stimuli. The adaptive immune response requires prior exposure to microbial antigens, is mediated primarily by CD4 + T-lymphocytes and serves to further amplify acute inflammation. Although acute inflammation is fundamentally beneficial, severe inflammation can precipitate the systemic inflammatory response syndrome. This syndrome is characterized by hyperinflammation and can cause organ injury, shock and death in its most severe forms. Overall, our understanding of inflammation has increased tremendously during the past 20 years. However, these basic science advances have not yet translated into widespread benefit for patients suffering from trauma, sepsis and systemic inflammation.


The Hospital for the Ruptured and Crippled Eugene H. Pool, fourth Surgeon-in-Chief 1933-1935 followed by Philip D. Wilson, fifth Surgeon-in-Chief 1935.
In 1933, for the second time in the history of the Hospital for the Ruptured and Crippled (R & C), a general surgeon, Eugene Hillhouse Pool, MD, was appointed Surgeon-in-Chief by the Board of Managers of the New York Society for the Relief of the Ruptured and Crippled. R & C (whose name was changed to the Hospital for Special Surgery in 1940), then the oldest orthopaedic hospital in the country, was losing ground as the leading orthopaedic center in the nation. The R & C Board charged Dr. Pool with the task of recruiting the nation's best orthopaedic surgeon to become the next Surgeon-in-Chief. Phillip D. Wilson, MD, from the Massachusetts General Hospital in Boston and the Harvard Medical School was selected and agreed to accept this challenge. He joined the staff of the Hospital for the Ruptured and Crippled in the spring of 1934 as Director of Surgery and replaced Dr. Pool as Surgeon-in-Chief the next year. It was the time of the Great Depression, which added a heavy financial toll to the daily operations of the hospital. With a clear and courageous vision, Dr. Wilson reorganized the hospital, its staff responsibilities, professional education and care of patients. He established orthopaedic fellowships to support young orthopaedic surgeons interested in conducting research and assisted them with the initiation of their new practices. Recognizing that the treatment of crippling conditions and hernia were becoming separate specialties, one of his first decisions was to restructure the Hernia Department to become the General Surgery Department. His World War I experiences in Europe helped develop his expertise in the fields of fractures, war trauma and amputations, providing a broad foundation in musculoskeletal diseases that was to be beneficial to him in his future role as the leader of R & C.


[Mesenteric infarct--when is the vascular surgeon needed?].
The treatment of acute mesenteric infarction is a therpeutical challenge. The mortality rate of these, mainly beyond 70 year old patients has remained extraordinary high despite diagnostic and therapeutic efforts and progress in intensive care medicine. Additionally to the initial hypoxaemia of the bowel a more severe tissue damage is induced via the "reperfusion injury", with increase of the vital risk. Besides the detection of occlusive vascular lesions of different etiology, which have to be treated immediately, "non occlusive mesenteric ischemia" can be identified. Visceral biplane angiography is the diagnostic tool of choice, giving basic informations on concomitant lesions, for a rational vascular surgical intervention and additionally the opportunity to administer vasoactive drugs. Despite increasing treatment modalities influencing the severity of the "reperfusion injury" the most important criteria for the patients' survival is the early diagnosis and immediate treatment. Therefore, close cooperation of general practitioners, internists, radiologists, angiologists, general surgeons and vascular surgeons is required. The support of a vascular surgeon is necessary for complex arterial reconstructions as success and patency rate are significantly influenced by experience. Reocclusion and "redo"-surgery increase the risk substantially.


[Toxic epidermal necrolysis after the use of intermediate dose of cytosine arabinoside].
Toxic epidermal necrolysis is a drug-induced dermatologic disease related to Lyell syndrome, erythema multiforme and Stevens-Johnson syndrome. To report a fatal case of toxic epidermal necrolysis owing to intermediate dose of cytarabine. A 16 year-old female patient with acute lymphocytic leukemia (LLA-L1) treated with the Protocol of the Brazilian Group for Treatment of Leukemia of Childwood (GBTLI-85-AR). On the second day after the administration of intermediate dose of cytarabine (1.5 g/m2 i.v. every 12 hours for 3 days), she presented bullous lesions in the left buttock that disseminated envolving to necrosis, sepsis, and death on the 13th day. Cytarabine is frequently associated with dermatologic toxicity but, until now, there is no other case of toxic epidermal necrolysis described.


Metabolomic fingerprint reveals that metformin impairs one-carbon metabolism in a manner similar to the antifolate class of chemotherapy drugs.
Metabolomic fingerprint of breast cancer cells treated with the antidiabetic drug metformin revealed a significant accumulation of 5-formimino-tetrahydrofolate, one of the tetrahydrofolate forms carrying activated one-carbon units that are essential for the de novo synthesis of purines and pyrimidines. De novo synthesis of glutathione, a folate-dependent pathway interconnected with one-carbon metabolism was concomitantly depleted in response to metformin. End-product reversal studies demonstrated that thymidine alone leads to a significant but incomplete protection from metformin's cytostatic effects. The addition of the substrate hypoxanthine for the purine salvage pathway produces major rightward shifts in metformin's growth inhibition curves. Metformin treatment failed to activate the DNA repair protein ATM kinase and the metabolic tumor suppressor AMPK when thymidine and hypoxanthine were present in the extracellular milieu. Our current findings suggest for the first time that metformin can function as an antifolate chemotherapeutic agent that induces the ATM/AMPK tumor suppressor axis secondarily following the alteration of the carbon flow through the folate-related one-carbon metabolic pathways.


Development of digital mirror therapy for stroke-severe patients.
Mirror therapy (MT), which is used in the existing stroke rehabilitation environment, has significant limitations for use with severe stroke patients. Since mirrors only reflect symmetrical movement, allowing a patient to observe precise asymmetrical movement is impossible. This study proposes a new MT system by developing a pyramid hologram technology that uses delayed motion to create realistic images. Significant differences, observed via electroencephalogram, were shown in all motor cortex channels immediately after the event in the delayed condition when compared to before the event (C3: p < 0.001; Cz: p < 0.001, C4: p < 0.001). The illusion of asymmetrical movement using the proposed system can be applied to severe stroke patients to increase the positive outcome of rehabilitation.


Opportunities for promoting youth physical activity: an examination of youth summer camps.
Youth summer camp programs have the potential to provide opportunities for physical activity, but little to no research has been conducted to determine activity levels of campers. This study aimed to examine physical activity occurring in day and resident summer camps and how activity levels differed in these camps based upon demographic characteristics. Pedometer data were collected during hours of camp operation from 150 day campers and 114 resident campers between the ages of 8 and 12 years old. Independent t tests were used to compare physical activity by sex, race, and Body Mass Index. Campers at day camps averaged 11,916 steps per camp day, while resident campers averaged 19,699 steps per camp day. Day campers averaged 1586 steps per hour over 7.5 hour days and resident campers averaged 1515 steps per hour over 13 hour days. Male sex, Caucasian race, and normal Body Mass Index were significant correlates of more physical activity. Youth summer camps demonstrate the potential to provide ample opportunities for physical activity during the summer months. Traditional demographic disparities persisted in camps, but the structure of camp programs should allow for changes to increase physical activity for all participants.


Distal hyperintense vessels on FLAIR: an MRI marker for collateral circulation in acute stroke?
Hyperintense vessels (HV) on fluid-attenuated inversion recovery imaging are frequently observed in acute ischemic stroke patients. However, the exact mechanism and clinical implications of this sign have not yet been clearly defined. The features of HV and its relevance to other imaging factors are presented here. Prominence and location of HV were documented in 52 consecutive patients with middle cerebral artery (MCA) territory infarction, before treatment with IV recombinant tissue plasminogen activator. Pretreatment ischemic lesion volume, perfusion lesion volume, and vessel occlusion were determined in addition to recanalization status and ischemic lesion volume on follow-up imaging. NIH Stroke Scale (NIHSS) was used as a measure of clinical severity. HV distal to arterial occlusion was observed in 73% of patients; more frequent in proximal than distal MCA occlusion patients. Among the 38 patients with proximal MCA occlusion, initial perfusion lesion volume was comparable among patients with different grade distal HV. However, patients with more prominent distal HV had smaller initial, 24-hour, and subacute ischemic lesion volumes and lower initial NIHSS scores. The presence of distal hyperintense vessels before thrombolytic treatment is associated with large diffusion-perfusion mismatch and smaller subacute ischemic lesion volumes in patients with proximal middle cerebral artery occlusion. DWI = diffusion-weighted imaging; FLAIR = fluid-attenuated inversion recovery; GRE = gradient recalled echo; HV = hyperintense vessels; MCA = middle cerebral artery; MRA = magnetic resonance angiography; MTT = mean transit time; NIHSS = NIH Stroke Scale; PWI = perfusion-weighted imaging; rt-PA = recombinant tissue plasminogen activator; TE = echo time; TI = inversion time; TIMI = thrombolysis in myocardial infarction; TR = repetition time.


Macrofilaricidal efficacy of single and repeated oral and subcutaneous doses of flubendazole in Litomosoides sigmodontis infected jirds.
Flubendazole (FBZ) is highly efficacious against filarial nematodes after parenteral administration and presents a promising macrofilaricidal drug candidate for the elimination of onchocerciasis and other filariae. In the present study the efficacy of a newly developed bioavailable amorphous solid dispersion (ASD) oral formulation of FBZ was investigated in the Litomosoides sigmodontis jird model. FBZ was administered to chronically infected, microfilariae-positive jirds by single (40mg/kg), repeated (2, 6 or 15mg/kg for 5 or 10 days) oral (OR) doses or single subcutaneous (SC) injections (2 or 10mg/kg). Jirds treated with 5 SC injections at 10mg/kg served as positive controls, with untreated animals used as negative controls. After OR doses, FBZ is rapidly absorbed and cleared and the exposures increased dose proportionally. SC administered FBZ was slowly released from the injection site and plasma levels remained constant up to necropsy eight weeks after treatment end. Increasing single SC doses caused less than dose-proportional exposures. At necropsy, all animals receiving 1x or 5x 10mg/kg SC FBZ had cleared all adult worms and the 1x 2mg/kg SC treatment had reduced the adult worm burden by 98%. 10x 15mg/kg OR FBZ reduced the adult worm burden by 95%, whereas 1x 40mg/kg and 5x 15mg/kg OR reduced the worm burden by 85 and 84%, respectively. Microfilaremia was completely cleared at necropsy in all animals of the SC treatment regimens, while all oral FBZ treatment regimens reduced the microfilaremia by >90% in a dose and duration dependent manner. In accordance, embryograms from female worms revealed a FBZ dose and duration dependent inhibition of embryogenesis. Histological analysis of the remaining female adult worms showed that FBZ had damaged the body wall, intestine and most prominently the uterus and uterine content. Results of this study demonstrate that single and repeated SC injections and repeated oral administrations of FBZ have an excellent macrofilaricidal effect.


Oestrogen-receptor-positive breast cancer: towards bridging histopathological and molecular classifications.
The oestrogen receptor (ER) pathway is key for survival and progression in a significant proportion of breast cancers. The ER can be activated by oestrogen or activated due to "crosstalk" with growth factor receptor pathways. Activated ER signals through transcriptional and non-transcriptional mechanisms. Immunohistochemistry (IHC), in spite of the shortcomings, remains the method of choice as it provides for in situ assessment of ER expression within the tumour cells. This capability is lost in tissue grinding methods that assess oestrogen-binding activity or messenger RNAs in tumours. IHC is also not influenced by the presence of non-tumoural cells or low amounts of tumour cells within samples examined. It is clear that ER-positive tumours do not represent a single entity. Irrespective of the terminology used, low-grade ER-positive (also known as luminal A) tumours need to be differentiated from high-grade/highly proliferative ER-positive tumours. This can be done in a variety of ways including but not limited to analysis of FOXA1 and GATA-3 by IHC, and limited molecular profiling by Oncotype DX, MGH2-gene signature, intrinsic gene signature or MapQuant Dx. Several areas of ER biology are still poorly understood; these include: its function in the cytoplasm/plasma membrane, its role in the differentiation to proliferation switch, and pathways associated with resistance to hormonal therapy. A detailed understanding of these areas will permit better classification and a personalised approach to management of ER-positive breast cancers.


The plastid ndh genes code for an NADH-specific dehydrogenase: isolation of a complex I analogue from pea thylakoid membranes.
The plastid genomes of several plants contain ndh genes-homologues of genes encoding subunits of the proton-pumping NADH:ubiquinone oxidoreductase, or complex I, involved in respiration in mitochondria and eubacteria. From sequence similarities with these genes, the ndh gene products have been suggested to form a large protein complex (Ndh complex); however, the structure and function of this complex remains to be established. Herein we report the isolation of the Ndh complex from the chloroplasts of the higher plant Pisum sativum. The purification procedure involved selective solubilization of the thylakoid membrane with dodecyl maltoside, followed by two anion-exchange chromatography steps and one size-exclusion chromatography step. The isolated Ndh complex has an apparent total molecular mass of approximately 550 kDa and according to SDS/PAGE consists of at least 16 subunits including NdhA, NdhI, NdhJ, NdhK, and NdhH, which were identified by N-terminal sequencing and immunoblotting. The Ndh complex showed an NADH- and deamino-NADH-specific dehydrogenase activity, characteristic of complex I, when either ferricyanide or the quinones menadione and duroquinone were used as electron acceptors. This study describes the isolation of the chloroplast analogue of the respiratory complex I and provides direct evidence for the function of the plastid Ndh complex as an NADH:plastoquinone oxidoreductase. Our results are compatible with a dual role for the Ndh complex in the chlororespiratory and cyclic photophosphorylation pathways.


Physician bonuses: tarnished awards?
Physician bonuses tied to meeting a managed care plan's profit or utilization targets reward doctors for stinting on care. Bonuses tied to patient satisfaction surveys measure service levels, not quality. Still, these deeply flawed payment systems are becoming more widespread.


Efferent connections of the parvalbumin-positive (PV1) nucleus in the lateral hypothalamus of rodents.
A solitary cluster of parvalbumin-positive neurons--the PV1 nucleus--has been observed in the lateral hypothalamus of rodents. In the present study, we mapped the efferent connections of the PV1 nucleus using nonspecific antero- and retrograde tracers in rats, and chemoselective, Cre-dependent viral constructs in parvalbumin-Cre mice. In both species, the PV1 nucleus was found to project mainly to the periaqueductal grey matter (PAG), predominantly ipsilaterally. Indirectly in rats and directly in mice, a discrete, longitudinally oriented cylindrical column of terminal fields (PV1-CTF) was identified ventrolateral to the aqueduct on the edge of the PAG. The PV1-CTF is particularly dense in the rostral portion, which is located in the supraoculomotor nucleus (Su3). It is spatially interrupted over a short stretch at the level of the trochlear nucleus and abuts caudally on a second parvalbumin-positive (PV2) nucleus. The rostral and the caudal portions of the PV1-CTF consist of axonal endings, which stem from neurons scattered throughout the PV1 nucleus. Topographically, the longitudinal orientation of the PV1-CTF accords with that of the likewise longitudinally oriented functional modules of the PAG, but overlaps none of them. Minor terminal fields were identified in a crescentic column of the lateral PAG, as well as in the Edinger-Westphal, the lateral habenular, and the laterodorsal tegmental nuclei. So far, no obvious functions have been attributed to this small, circumscribed column ventrolateral to the aqueduct, the prime target of the PV1 nucleus.


Effect of vertical banded gastroplasty on hypertension, diabetes and dyslipidemia.
Operations for morbid obesity that are effective in inducing weight loss improve blood pressure, glycemic control and dyslipidemia. Our purpose was to study the effectiveness of VBG in improving hypertension, diabetes and dyslipidemias in morbidly obese patients. Retrospective analysis of a cohort of 80 morbidly, obese patients was conducted. Characteristics of patients were: mean age 37 years; gender: women 65, men 15; mean initial BMI 49.5 kg/m2; follow-up > 5 years in 52 patients. Blood pressure, cholesterol and triglyceride levels and glycemia were assessed preoperatively and 1, 3, 6, 12, 18 months and every year after VBG. Hypertension resolved in 65.5% of patients (38 of 58), serum cholesterol level became normal in 34.28% of patients (12 of 35), triglyceride level became normal in 77.77% (7 of 9) and diabetes resolved in 55.55% (5 of 9). Weight loss induced by VBG improves blood pressure, triglyceride levels and glycemia. The beneficial changes occur early in the postoperative period and last 5 years or more, despite of the trend to regain some weight. Improvements in risk factors were greater in patients with higher preoperative values.


Yolk protein is expressed in the insect testis and interacts with sperm.
Male and female gametes follow diverse developmental pathways dictated by their distinct roles in fertilization. While oocytes of oviparous animals accumulate yolk in the cytoplasm, spermatozoa slough off most of their cytoplasm in the process of individualization. Mammalian spermatozoa released from the testis undergo extensive modifications in the seminal ducts involving a variety of glycoproteins. Ultrastructural studies suggest that glycoproteins are involved in sperm maturation in insects; however, their characterization at the molecular level is lacking. We reported previously that the circadian clock controls sperm release and maturation in several insect species. In the moth, Spodoptera littoralis, the secretion of glycoproteins into the seminal fluid occurs in a daily rhythmic pattern. The purpose of this study was to characterize seminal fluid glycoproteins in this species and elucidate their role in the process of sperm maturation. We collected seminal fluid proteins from males before and after daily sperm release. These samples were separated by 2-D gel electrophoresis, and gels were treated with a glycoprotein-detecting probe. We observed a group of abundant glycoproteins in the sample collected after sperm release, which was absent in the sample collected before sperm release. Sequencing of these glycoproteins by mass spectroscopy revealed peptides bearing homology with components of yolk, which is known to accumulate in developing oocytes. This unexpected result was confirmed by Western blotting demonstrating that seminal fluid contains protein immunoreactive to antibody against yolk protein YP2 produced in the follicle cells surrounding developing oocytes. We cloned the fragment of yp2 cDNA from S. littoralis and determined that it is expressed in both ovaries and testes. yp2 mRNA and YP2 protein were detected in the somatic cyst cells enveloping sperm inside the testis. During the period of sperm release, YP2 protein appears in the seminal fluid and forms an external coat on spermatozoa. One of the yolk protein precursors YP2, which in females accumulate in the oocytes to provision developing embryos, appears to have a second male-specific role. It is produced in the testes and released into the seminal fluid where it interacts with sperm. These data reveal unexpected common factor in the maturation of insect eggs and sperm.


Expression and Functional Analysis of the Compact Thermophilic Anoxybacillus flavithermus Cas9 Nuclease.
Research on Cas9 nucleases from different organisms holds great promise for advancing genome engineering and gene therapy tools, as it could provide novel structural insights into CRISPR editing mechanisms, expanding its application area in biology and medicine. The subclass of thermophilic Cas9 nucleases is actively expanding due to the advances in genome sequencing allowing for the meticulous examination of various microorganisms' genomes in search of the novel CRISPR systems. The most prominent thermophilic Cas9 effectors known to date are GeoCas9, ThermoCas9, IgnaviCas9, AceCas9, and others. These nucleases are characterized by a varying temperature range of the activity and stringent PAM preferences; thus, further diversification of the naturally occurring thermophilic Cas9 subclass presents an intriguing task. This study focuses on generating a construct to express a compact Cas9 nuclease (AnoCas9) from the thermophilic microorganism Anoxybacillus flavithermus displaying the nuclease activity in the 37-60 °C range and the PAM preference of 5'-NNNNCDAA-3' in vitro. Here, we highlight the close relation of AnoCas9 to the GeoCas9 family of compact thermophilic Cas9 effectors. AnoCas9, beyond broadening the repertoire of Cas9 nucleases, suggests application in areas requiring the presence of thermostable CRISPR/Cas systems in vitro, such as sequencing libraries' enrichment, allele-specific isothermal PCR, and others.