Postextubation dysphagia is persistent and associated with poor outcomes in survivors of critical illness.
Dysphagia is common among survivors of critical illness who required mechanical ventilation during treatment. The risk factors associated with the development of postextubation dysphagia, and the effects of dysphagia on patient outcomes, have been relatively unexplored. We conducted a retrospective, observational cohort study from 2008 to 2010 of all patients over 17 years of age admitted to a university hospital ICU who required mechanical ventilation and subsequently received a bedside swallow evaluation (BSE) by a speech pathologist. A BSE was performed after mechanical ventilation in 25% (630 of 2,484) of all patients. After we excluded patients with stroke and/or neuromuscular disease, our study sample size was 446 patients. We found that dysphagia was present in 84% of patients (n = 374) and classified dysphagia as absent, mild, moderate or severe in 16% (n = 72), 44% (n = 195), 23% (n = 103) and 17% (n = 76), respectively. In univariate analyses, we found that statistically significant risk factors for severe dysphagia included long duration of mechanical ventilation and reintubation. In multivariate analysis, after adjusting for age, gender and severity of illness, we found that mechanical ventilation for more than seven days remained independently associated with moderate or severe dysphagia (adjusted odds ratio (AOR) = 2.84 [interquartile range (IQR) = 1.78 to 4.56]; P < 0.01). The presence of severe postextubation dysphagia was significantly associated with poor patient outcomes, including pneumonia, reintubation, in-hospital mortality, hospital length of stay, discharge status and surgical placement of feeding tubes. In multivariate analysis, we found that the presence of moderate or severe dysphagia was independently associated with the composite outcome of pneumonia, reintubation and death (AOR = 3.31 [IQR = 1.89 to 5.90]; P < 0.01). In a large cohort of critically ill patients, long duration of mechanical ventilation was independently associated with postextubation dysphagia, and the development of postextubation dysphagia was independently associated with poor patient outcomes.


Evaluating the Association Between the Three Different Ejection Fraction Measurement Techniques and Left Ventricle Global Strain.
The prognosis of cardiovascular diseases (CVDs) is directly associated with systolic function based on the measurement of ejection fraction (EF), and many studies have indicated that the left ventricular global strain (LVGS) provides better predictivity than the EF measurement in the diagnosis, prognosis, survival, and CVD staging. However, these studies did not investigate the correlation between the EF measurement and the LVGS parameters, or which parameters are better correlated with LVGS, but we analyzed the association between three EF measurement methods and LVGS. This study included 62 patients that applied to the clinic between October 2015 and March 2016. An echocardiography examination of these patients was performed. The exclusion criteria were atrial fibrillation and suboptimal image quality. Sixty-two patients (the average age 61.0±12.6 years; 56% male and 44% female) were enrolled in the study. A statistically significant association was found between the visual EF and Simpson EF measurements and the LVGS parameters (p<0.001). While the visual EF was moderately correlated with the LVGS parameters (r=0.44), there was a good correlation between the Simpson EF and the LVGS parameters (r=0.710). In this study, we demonstrate that the Simpson's rule LVEF correlates better with LVGS than the Teicholtz method or visual EF and that it has a better area under the curve value for determining an abnormal LVGS. Therefore, we recommend the use of the Simpson EF for the EF measurement that has a better correlation with the LVGS values in the patients whose ventricle functions should be evaluated.


Electric Field Controlled Self-Assembly of Hierarchically Ordered Membranes.
Self-assembly in the presence of external forces is an adaptive, directed organization of molecular components under nonequilibrium conditions. While forces may be generated as a result of spontaneous interactions among components of a system, intervention with external forces can significantly alter the final outcome of self-assembly. Superimposing these intrinsic and extrinsic forces provides greater degrees of freedom to control the structure and function of self-assembling materials. In this work we investigate the role of electric fields during the dynamic self-assembly of a negatively charged polyelectrolyte and a positively charged peptide amphiphile in water leading to the formation of an ordered membrane. In the absence of electric fields, contact between the two solutions of oppositely charged molecules triggers the growth of closed membranes with vertically oriented fibrils that encapsulate the polyelectrolyte solution. This process of self-assembly is intrinsically driven by excess osmotic pressure of counterions, and the electric field is found to modify the kinetics of membrane formation, and also its morphology and properties. Depending on the strength and orientation of the field we observe a significant increase or decrease of up to nearly 100% in membrane thickness, as well as the controlled rotation of nanofiber growth direction by 90 degrees, resulting in a significant increase in mechanical stiffness. These results suggest the possibility of using electric fields to control structure in self-assembly processes involving diffusion of oppositely charged molecules.


The TNF-derived TIP peptide activates the epithelial sodium channel and ameliorates experimental nephrotoxic serum nephritis.
In mice, the initial stage of nephrotoxic serum-induced nephritis (NTN) mimics antibody-mediated human glomerulonephritis. Local immune deposits generate tumor necrosis factor (TNF), which activates pro-inflammatory pathways in glomerular endothelial cells (GECs) and podocytes. Because TNF receptors mediate antibacterial defense, existing anti-TNF therapies can promote infection; however, we have previously demonstrated that different functional domains of TNF may have opposing effects. The TIP peptide mimics the lectin-like domain of TNF, and has been shown to blunt inflammation in acute lung injury without impairing TNF receptor-mediated antibacterial activity. We evaluated the impact of TIP peptide in NTN. Intraperitoneal administration of TIP peptide reduced inflammation, proteinuria, and blood urea nitrogen. The protective effect was blocked by the cyclooxygenase inhibitor indomethacin, indicating involvement of prostaglandins. Targeted glomerular delivery of TIP peptide improved pathology in moderate NTN and reduced mortality in severe NTN, indicating a local protective effect. We show that TIP peptide activates the epithelial sodium channel(ENaC), which is expressed by GEC, upon binding to the channel's α subunit. In vitro, TNF treatment of GEC activated pro-inflammatory pathways and decreased the generation of prostaglandin E2 and nitric oxide, which promote recovery from NTN. TIP peptide counteracted these effects. Despite the capacity of TIP peptide to activate ENaC, it did not increase mean arterial blood pressure in mice. In the later autologous phase of NTN, TIP peptide blunted the infiltration of Th17 cells. By countering the deleterious effects of TNF through direct actions in GEC, TIP peptide could provide a novel strategy to treat glomerular inflammation.


Oral care in cancer nursing: Practice and barriers.
Oral health awareness and oral care are crucial aspects of oncology nursing practice. However, very few studies concentrate on the oral care of cancer patients undergoing cancer treatment and nursing practice in the Indian subcontinent scenario/situation. Most of the published studies have been conducted in the Western and European countries. This study aimed to determine the nurses' practice and barriers regarding oral care in cancer patients undergoing chemotherapy and radiation therapy. A cross-sectional descriptive survey was conducted among 158 staff nurses working in oncology-related areas from four different hospitals of Dakshina Kannada district and Udupi district of Karnataka state, India. Descriptive and inferential statistics was used by using SPSS 16 version. More than half of respondents [54 (34.2%)] did not perform oral care as a part of routine duties. Maintenance of various records, lack of manpower, and lack of standard operating procedures were major barriers in providing oral care. Documentation audit revealed that nurses recorded oral care in the chart only when order was present in the care sheet, but oral problem assessment was not recorded at all. In all four hospitals surveyed, there was no protocol specifically designed for oral care of cancer patients. Nurses expressed that oral care in cancer patients was one of the most ignored aspect in oncology nursing. Our result highlights the need to develop evidence-based oral care intervention protocol and motivate staff nurses to attend continuing nursing educations regularly to keep themselves abreast of the latest trends in order to render comprehensive care to the patients.


Chemical Genomics Approach Leads to the Identification of Hesperadin, an Aurora B Kinase Inhibitor, as a Broad-Spectrum Influenza Antiviral.
Influenza viruses are respiratory pathogens that are responsible for annual influenza epidemics and sporadic influenza pandemics. Oseltamivir (Tamiflu®) is currently the only FDA-approved oral drug that is available for the prevention and treatment of influenza virus infection. However, its narrow therapeutic window, coupled with the increasing incidence of drug resistance, calls for the next generation of influenza antivirals. In this study, we discovered hesperadin, an aurora B kinase inhibitor, as a broad-spectrum influenza antiviral through forward chemical genomics screening. Hesperadin inhibits multiple human clinical isolates of influenza A and B viruses with single to submicromolar efficacy, including oseltamivir-resistant strains. Mechanistic studies revealed that hesperadin inhibits the early stage of viral replication by delaying the nuclear entry of viral ribonucleoprotein complex, thereby inhibiting viral RNA transcription and translation as well as viral protein synthesis. Moreover, a combination of hesperadin with oseltamivir shows synergistic antiviral activity, therefore hesperadin can be used either alone to treat infections by oseltamivir-resistant influenza viruses or used in combination with oseltamivir to delay resistance evolution among oseltamivir-sensitive strains. In summary, the discovery of hesperadin as a broad-spectrum influenza antiviral offers an alternative to combat future influenza epidemics and pandemics.


Treatment of arrhythmias by external charged particle beams: a Langendorff feasibility study.
Hadron therapy has already proven to be successful in cancer therapy, and might be a noninvasive alternative for the ablation of cardiac arrhythmias in humans. We present a pilot experiment investigating acute effects of a 12C irradiation on the AV nodes of porcine hearts in a Langendorff setup. This setup was adapted to the requirements of charged particle therapy. Treatment plans were computed on calibrated CTs of the hearts. Irradiation was applied in units of 5 and 10 Gy over a period of about 3 h until a total dose of up to 160 Gy was reached. Repeated application of the same irradiation field helped to mitigate motion artifacts in the resulting dose distribution. After irradiation, PET scans were performed to verify accurate dose application. Acute AV blocks were identified. No other acute effects were observed. Hearts were kept in sinus rhythm for up to 6 h in the Langendorff setup. We demonstrated that 12C ions can be used to select a small target in the heart and, thereby, influence the electrical conduction system. Second, our pilot study seems to suggest that no adverse effects have to be expected immediately during heavy ion irradiation in performing subsequent experiments with doses of 30-60 Gy and intact pigs.


MALDI-TOF MS as an innovative tool for detection of Plasmodium parasites in Anopheles mosquitoes.
Malaria is still a major public health issue worldwide, and one of the best approaches to fight the disease remains vector control. The current methods for mosquito identification include morphological methods that are generally time-consuming and require expertise, and molecular methods that require laboratory facilities with relatively expensive running costs. Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) technology, routinely used for bacterial identification, has recently emerged in the field of entomology. The aim of the present study was to assess whether MALDI-TOF MS could successfully distinguish Anopheles stephensi mosquitoes according to their Plasmodium infection status. C57BL/6 mice experimentally infected with Plasmodium berghei were exposed to An. stephensi bites. For the determination of An. stephensi infection status, mosquito cephalothoraxes were dissected and submitted to mass spectrometry analyses and DNA amplification for molecular analysis. Spectra were grouped according to mosquitoes' infection status and spectra quality was validated based on intensity and reproducibility within each group. The in-lab MALDI-TOF MS arthropod reference spectra database, upgraded with representative spectra from both groups (infected/non-infected), was subsequently queried blindly with cephalothorax spectra from specimens of both groups. The MALDI TOF MS profiles generated from protein extracts prepared from the cephalothorax of An. stephensi allowed distinction between infected and uninfected mosquitoes. Correct classification was obtained in blind test analysis for (79/80) 98.75% of all mosquitoes tested. Only one of 80 specimens, an infected mosquito, was misclassified in the blind test analysis. Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry appears to be a promising, rapid and reliable tool for the epidemiological surveillance of Anopheles vectors, including their identification and their infection status.


Effects of volume on outcome in hepatobiliary surgery: a review with guidelines proposal.
The positive relationship between volume and outcome in hepatobiliary surgery has been demonstrated for many years. As for other complex surgical procedures, both improved short- and long-term outcomes have been associated with a higher volume of procedures. However, whether the centralization of complex hepatobiliary procedures makes full sense because it should be associated with higher quality of care, as reported in the literature, precise criteria on what to centralize, where to centralize, and who should be entitled to perform complex procedures are still missing. Indeed, despite the generalized consensus on centralization in hepatobiliary surgery, this topic remains very complex because many determinants are involved in such a centralization process, of which some of them cannot be easily controlled. In the context of different health systems worldwide, such as national health systems and private insurance, there are different stakeholders that demand different needs: politicians, patients, surgeons, institutions and medical associations do not always have the same needs. Starting from a review of the literature on centralization in hepatobiliary surgery, we will propose some guidelines that, while not data-driven due to low evidence in the literature, will be based on good clinical practice.


Linear Ubiquitin Chains: Cellular Functions and Strategies for Detection and Quantification.
Ubiquitination of proteins is a sophisticated post-translational modification implicated in the regulation of an ever-growing abundance of cellular processes. Recent insights into different layers of complexity have shaped the concept of the ubiquitin code. Key players in determining this code are the number of ubiquitin moieties attached to a substrate, the architecture of polyubiquitin chains, and post-translational modifications of ubiquitin itself. Ubiquitination can induce conformational changes of substrates and alter their interactive profile, resulting in the formation of signaling complexes. Here we focus on a distinct type of ubiquitination that is characterized by an inter-ubiquitin linkage through the N-terminal methionine, called M1-linked or linear ubiquitination. Formation, recognition, and disassembly of linear ubiquitin chains are highly specific processes that are implicated in immune signaling, cell death regulation and protein quality control. Consistent with their role in influencing signaling events, linear ubiquitin chains are formed in a transient and spatially regulated manner, making their detection and quantification challenging.


Unconventional microbial systems for the cost-efficient production of high-quality protein therapeutics.
Both conventional and innovative biomedical approaches require cost-effective protein drugs with high therapeutic potency, improved bioavailability, biocompatibility, stability and pharmacokinetics. The growing longevity of the human population, the increasing incidence and prevalence of age-related diseases and the better comprehension of genetic-linked disorders prompt to develop natural and engineered drugs addressed to fulfill emerging therapeutic demands. Conventional microbial systems have been for long time exploited to produce biotherapeutics, competing with animal cells due to easier operation and lower process costs. However, both biological platforms exhibit important drawbacks (mainly associated to intracellular retention of the product, lack of post-translational modifications and conformational stresses), that cannot be overcome through further strain optimization merely due to physiological constraints. The metabolic diversity among microorganisms offers a spectrum of unconventional hosts, that, being able to bypass some of these weaknesses, are under progressive incorporation into production pipelines. In this review we describe the main biological traits and potentials of emerging bacterial, yeast, fungal and microalgae systems, by comparing selected leading species with well established conventional organisms with a long run in protein drug production.


First Report of Colletotrichum sansevieriae Causing Anthracnose of Sansevieria trifasciata in Florida.
Sansevieria Thunberg, a member of the Agavaceae, contains around 60 species indigenous to Africa, Arabia, and India. Several species and their cultivars are commercially produced for use as interior and landscape foliage plants. During August 2010, several local nurseries submitted Sansevieria trifasciata samples to the Florida Extension Plant Diagnostic Clinic in Homestead. Leaves had round, water-soaked lesions and as the disease progressed, lesions rapidly enlarged and coalesced, resulting in severe leaf blight. Both young and mature leaves were affected. Closer examination of mature lesions revealed numerous brownish black acervuli that were produced in concentric rings, which is characteristic of anthracnose. The fungus was identified as Colletotrichum sansevieriae Nakamura based on typical cultural characteristics, conidial and appressoria morphology (1). Conidia were straight, cylindrical, obtuse at the apex, slightly acute at the base with a truncate attachment point, and 12.5 to 33 (18.4) × 4 to 8.9 (6.5) μm (n = 50). Hyphopodia were ovate, dark brown, single celled, and 6.2 to 8.7 (7.7) × 6.3 to 7.5 (7.3) μm (n = 25). Colonies on potato dextrose agar (PDA) were grayish white, felted with aerial mycelium, reverse gray to dark olivaceous gray, and partly cream in color. Sequences of the rDNA internal transcribed spacer (ITS) regions of two isolates (GenBank Accession Nos. JF911349 and JF911350) exhibited 99% nucleotide identity to an isolate of C. sansevieriae (GenBank Accession No. HQ433226) collected from diseased sansevieria in Australia. In addition, a maximum parsimony analysis (MEGA v.5.0) indicates that the two C. sansevieriae isolates from Florida are monophyletic (86% bootstrap support) with the type species from Japan (SA-1-2 AB212991; SA-1-1 AB212990) and the Australian isolate. Pathogenicity of our sequenced isolates was evaluated in greenhouse experiments. Twelve- to fourteen-week-old sansevieria plants were inoculated with conidial suspensions (1 × 106 conidia/ml) of C. sansevieriae. Inoculum or autoclaved water was sprayed over the foliage until runoff. Four plants of each of two economically important cultivars, Laurentii and Moonshine, were sprayed per treatment and the experiment was repeated twice. Inoculated plants were placed in a greenhouse at 29°C with 70 to 85% relative humidity. Plants were observed for disease development, which occurred within 10 days of inoculation for both cultivars. No symptoms developed on the control plants. Foliar lesions closely resembled those observed in the affected nurseries. C. sansevieriae was consistently reisolated from symptomatic tissue collected from greenhouse experiments. On the basis of molecular phylogenetics and distinguishing morphological characters, Nakamura et al. erected C. sansevieriae as a novel species that appears to be restricted to the host sansevieria (1). To our knowledge, this is the first report of C. sansevieriae causing anthracnose of sansevieria in Florida. Reference: (1) M. Nakamura et al. J. Gen. Plant Pathol. 72:253, 2006.


Novel inhibitors of 17beta-hydroxysteroid dehydrogenase type 1: templates for design.
The 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) catalyze the interconversion between the oxidized and reduced forms of androgens and estrogens at the 17 position. The 17beta-HSD type 1 enzyme (17beta-HSD1) catalyzes the reduction of estrone (E1) to estradiol and is expressed in malignant breast cells. Inhibitors of this enzyme thus have potential as treatments for hormone dependent breast cancer. Syntheses and biological evaluation of novel non-steroidal inhibitors designed to mimic the E1 template are reported using information from potent steroidal inhibitors. Of the templates investigated biphenyl ethanone was promising and led to inhibitors with IC(50) values in the low micromolar range.


Budesonide and terbutaline or terbutaline alone in children with mild asthma: effects on bronchial hyperresponsiveness and diurnal variation in peak flow.
The effects of treatment with budesonide (200 micrograms twice daily) and terbutaline (500 micrograms four times daily) has been compared with the effects of placebo and terbutaline in 27 children with mild asthma, aged 7-14 years, in a double blind, randomised placebo controlled study over eight weeks. Bronchial responsiveness (PC20 histamine), lung function, the amplitude of diurnal variation in peak expiratory flow (PEF), and symptom scores were measured. Baseline FEV1 was over 70% predicted and PC20 histamine less than 8 mg/ml. Twelve children were treated with budesonide and terbutaline and 15 with placebo and terbutaline. After four and eight weeks of treatment the change in PC20 was significantly greater after budesonide and terbutaline than after terbutaline alone by 2.1 (95% CI 0.5-3.8) and 1.3 (95% CI 0.1-2.5) doubling doses respectively. Mean FEV1 did not change in either group. The change in afternoon and nocturnal PEF was significantly greater after budesonide and terbutaline than after terbutaline alone. The amplitude of diurnal variation in PEF did not change significantly in either group. Peak flow reversibility decreased in the budesonide group. There were no differences between treatments for cough and dyspnoea, but wheeze improved in the budesonide group. The children with mild asthma treated with budesonide and terbutaline showed improvement in bronchial responsiveness, afternoon and nocturnal PEF, and symptoms of wheeze and a fall in peak flow reversibility by comparison with those who received terbutaline alone.


Physiological and pathophysiological concentrations of fatty acids induce lipid droplet accumulation and impair functional performance of tissue engineered skeletal muscle.
Fatty acids (FA) exert physiological and pathophysiological effects leading to changes in skeletal muscle metabolism and function, however, in vitro models to investigate these changes are limited. These experiments sought to establish the effects of physiological and pathophysiological concentrations of exogenous FA upon the function of tissue engineered skeletal muscle (TESkM). Cultured initially for 14 days, C2C12 TESkM was exposed to FA-free bovine serum albumin alone or conjugated to a FA mixture (oleic, palmitic, linoleic, and α-linoleic acids [OPLA] [ratio 45:30:24:1%]) at different concentrations (200 or 800 µM) for an additional 4 days. Subsequently, TESkM morphology, functional capacity, gene expression and insulin signaling were analyzed. There was a dose response increase in the number and size of lipid droplets within the TESkM (p < .05). Exposure to exogenous FA increased the messenger RNA expression of genes involved in lipid storage (perilipin 2 [p < .05]) and metabolism (pyruvate dehydrogenase lipoamide kinase isozyme 4 [p < .01]) in a dose dependent manner. TESkM force production was reduced (tetanic and single twitch) (p < .05) and increases in transcription of type I slow twitch fiber isoform, myosin heavy chain 7, were observed when cultured with 200 µM OPLA compared to control (p < .01). Four days of OPLA exposure results in lipid accumulation in TESkM which in turn results in changes in muscle function and metabolism; thus, providing insight ito the functional and mechanistic changes of TESkM in response to exogenous FA.


The contractile basis of amoeboid movement. V. The control of gelation, solation, and contraction in extracts from Dictyostelium discoideum.
Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.


The H3 receptor agonist immepip does not affect l-dopa-induced abnormal involuntary movements in 6-OHDA-lesioned rats.
The treatment of dyskinesia in Parkinson׳s disease remains poor but H3 receptor agonists have been suggested as a novel pharmacological approach. We examined the effects of the H3 agonist, immepip, in 6-OHDA-lesioned rats exhibiting AIMs (abnormal involuntary movements), a rat analogue of dyskinesia, in response to l-dopa compared to the known anti-dyskinetic agents amantadine, MK-801 and 8-OHDPAT. We then attempted to extend these studies in to dyskinetic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated common marmosets. Amantadine, MK-801 and 8-OHDPAT all dose-dependently reduced l-dopa-induced axial, lingual and oral (ALO) AIMs in 6-OHDA-lesioned animals accompanied by a reduction in contralateral rotation with higher doses of amantadine and MK-801. By contrast, immepip had no effect on AIMs expression or contralateral rotation. In the MPTP-treated common marmoset exhibiting dyskinesia to l-dopa, immepip alone induced retching and in combination with l-dopa administered subcutaneously or orally induced the rapid onset of retching and vomiting which was not controlled by pretreatment with domperidone. Administration of the unrelated H3 agonist, imetit had the same effect. Despite causing negative side-effects, it appears that both agonists reduced the antiparkinsonian response to l-dopa resulting in reduced dyskinesia. H3 agonists appear unlikely candidates for the treatment of dyskinesia in PD based on lack of evidence of efficacy and potential adverse effects.


Excess deaths during the COVID-19 pandemic in Iran.
The actual number of deaths during the COVID-19 pandemic is expected to be higher than the reported deaths. We aimed to estimate the number of deaths in Iran during the COVID-19 pandemic from December 22, 2019 to March 20, 2022. We compared the number of age- and sex-specific deaths reported by Iran's Bureau of Vital Statistics with the predicted deaths estimated using an improved Lee-Carter model. We estimated the number of all-cause excess deaths in three scenarios, including the baseline scenario (without any undercounting of deaths) and 4% and 8% undercounting of all-cause deaths. We estimated 282,378 (95% confidence intervals [CI]: 225,439; 341,951) excess deaths in the baseline model. This number was 303,148 (95% CI: 246,417; 357,823) and 308,486 (95% CI: 250,607; 364,417) in the 4% and 8% scenarios, respectively. During the same period, Iran reported 139,610 deaths as being directly related to COVID-19. The ratio of reported COVID-19 deaths to total excess deaths ranged from 45.2% to 49.4% in the various scenarios. Most excess deaths occurred in the baseline scenario in males (157,552 [95% CI: 125,142; 191,265]) and those aged ≥75 years (102,369 [95% CI: 93,894; 111,188]). The reported number of COVID-19 deaths was less than half of Iran's estimated number of excess deaths. The results of this study will be helpful for health policymakers' planning, and call for strengthening the timeliness and accuracy of Iran's death registration systems, planning for more accurate monitoring of epidemics, and planning to provide support services for survivors' families.


Recognition of interspecies hybrid class I histocompatibility antigens by antigen-specific cytolytic T lymphocytes.
Two reciprocal interspecies hybrid class I histocompatibility genes have been constructed between genomic clones of human HLA-A2 and murine H-2Kb. The proteins encoded by these genes have been designated A21+2/Kb, where the polymorphic domains, alpha 1 and alpha 2, of HLA-A2 are linked to the carboxyl-terminal domains (alpha 3, transmembrane, and intracytoplasmic domains) of H-2Kb, and Kb1+2/A2, where the alpha 1 and alpha 2 domains of the H-2Kb antigen are linked to the carboxyl-terminal domains of HLA-A2. These genes have been transfected and expressed in recipient mouse L cells and human RD (rhabdomyosarcoma) cells. Both hybrid antigens were found to be serologically intact when tested with a panel of antigen-specific monoclonal antibodies. The monoclonal antibody W6/32, which recognizes a monomorphic determinant on all HLA-A, -B, and -C antigens, recognizes the alpha 1 and/or the alpha 2 domain, rather than the more conserved alpha 3 domain. Human cytolytic T lymphocytes (CTL) specific for the HLA-A2 antigen recognized the A2 and A21+2/Kb proteins only when expressed in human cells and not when expressed in mouse cells, even when surface antigen levels were 10-fold greater on the mouse cells than on the human cells. In contrast, a long-term, murine anti-H-2b CTL line not only lysed mouse L-cell lines that expressed the parental Kb and hybrid Kb1+2/A2 antigens but also lysed the Kb and Kb1+2/A2 human cell RD transformants as well. In both cases, the level of CTL recognition and lysis of the transformants that expressed the native antigen Kb was greater than of those transformants that expressed the hybrid antigen Kb1+2/A2. These data suggest that the carboxyl-terminal domains play some role in CTL allorecognition. The lack of human CTL recognition of HLA molecules expressed in mouse L cells, however, cannot be explained by the presence of a xenogeneic carboxyl terminus. Since murine CTL can recognize their target antigen when expressed on the surface of human cells, the possibility remains either that a ligand necessary for other molecular interactions of human CTL may be absent on mouse target cells or that murine and human CTL differ in affinity of binding to target antigens in the absence of accessory-molecule interactions.


Urinary pyridinoline cross-links as biomarkers of osteogenesis imperfecta.
Osteogenesis imperfecta (OI) is a group of genetic heterogeneous connective tissue disorders characterized by increased bone fragility and susceptibility to fractures. Laboratory diagnosis relies on time-consuming and cost-intensive biochemical and molecular genetics analyses. Therefore, it is desirable to identify and establish new diagnostic markers for OI that are reliable, cost-effective and easily accessible. In our study we have identified the ratio of the urinary pyridinoline cross-links lysyl-pyridinoline and hydroxylysyl-pyridinoline as a promising, time- and cost-effective biomarker for osteogenesis imperfecta, that could be used furthermore to investigate cases of suspected non-accidental injury in infants.


Arabidopsis immunity regulator EDS1 in a PAD4/SAG101-unbound form is a monomer with an inherently inactive conformation.
In plant innate immunity, enhanced disease susceptibility 1 (EDS1) integrates all pathogen-induced signals transmitted by TIR-type NLR receptors. Driven by an N-terminal α/β-hydrolase-fold domain with a protruding interaction helix, EDS1 assembles with two homologs, phytoalexin-deficient 4 (PAD4) and senescence-associated gene 101 (SAG101). The resulting heterodimers are critical for EDS1 function and structurally well characterized. Here, we resolve solution and crystal structures of unbound Arabidopsis thaliana EDS1 (AtEDS1) using nanobodies for crystallization. These structures, together with gel filtration and immunoprecipitation data, show that PAD4/SAG101-unbound AtEDS1 is stable as a monomer and does not form the homodimers recorded in public databases. Its PAD4/SAG101 anchoring helix is disordered unless engaged in protein/protein interactions. As in the complex with SAG101, monomeric AtEDS1 has a substrate-inaccessible esterase triad with a blocked oxyanion hole and without space for a covalent acyl intermediate. These new structures suggest that the AtEDS1 monomer represents an inactive or pre-activated ground state.


Rationale for co-targeting IGF-1R and ALK in ALK fusion-positive lung cancer.
Crizotinib, a selective tyrosine kinase inhibitor (TKI), shows marked activity in patients whose lung cancers harbor fusions in the gene encoding anaplastic lymphoma receptor tyrosine kinase (ALK), but its efficacy is limited by variable primary responses and acquired resistance. In work arising from the clinical observation of a patient with ALK fusion-positive lung cancer who had an exceptional response to an insulin-like growth factor 1 receptor (IGF-1R)-specific antibody, we define a therapeutic synergism between ALK and IGF-1R inhibitors. Similar to IGF-1R, ALK fusion proteins bind to the adaptor insulin receptor substrate 1 (IRS-1), and IRS-1 knockdown enhances the antitumor effects of ALK inhibitors. In models of ALK TKI resistance, the IGF-1R pathway is activated, and combined ALK and IGF-1R inhibition improves therapeutic efficacy. Consistent with this finding, the levels of IGF-1R and IRS-1 are increased in biopsy samples from patients progressing on crizotinib monotherapy. Collectively these data support a role for the IGF-1R-IRS-1 pathway in both ALK TKI-sensitive and ALK TKI-resistant states and provide a biological rationale for further clinical development of dual ALK and IGF-1R inhibitors.


Expression features of SOX9 associate with tumor progression and poor prognosis of hepatocellular carcinoma.
SOX9 as a member of the SOX (SRY [sex determining region Y] box) gene superfamily has been previously demonstrated to be a proto-oncogene in a variety of malignancies. However, the clinical significance of SOX9 expression in hepatocellular carcinoma (HCC) remains unclear. The aim of this study was to investigate the expression of SOX9 in HCC and determine its correlation with tumor progression and prognosis. One-hundred and thirty HCC patients who had undergone curative liver resection were selected and immunohistochemistry, Western blotting, and quantitative real time polymerase chain reaction (Q-PCR) were performed to analyze SOX9 expression in the respective tumors. Immunohistochemistry, Western blotting, and Q-PCR consistently confirmed SOX9 overexpression in HCC tissues compared with their adjacent nonneoplastic tissues (P ≪ 0.01). Additionally, immunostaining showed more SOX9 positive cells in the higher tumor stage (T3 ~ 4) and tumor grade (G3) than in the lower tumor stage (T1 ~ 2, P = 0.03) and tumor grade (G1 ~ 2, P = 0.01), respectively. Moreover, HCC patients with high SOX9 expression were significantly associated with lower 5-year overall survival (P ≪ 0.01) and lower 5-year disease-free survival (P ≪ 0.01), respectively. The Cox proportional hazards model further showed that SOX9 over-expression was an independent poor prognostic factor for both 5-year disease-free survival (hazards ratio [HR] = 2.621, 95% confidence interval[CI] = 1.548-5.829, P = 0.01) and 5-year overall survival (HR = 3.825, CI = 1.638-7.612, P = 0.003) in HCC. Our data suggest for the first time that the overexpression of SOX9 protein in HCC tissues is of predictive value on tumor progression and poor prognosis. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/9029740396926377.


Management and survival in advanced prostate cancer in Nairobi.
To evaluate the management and survival of patients with advanced prostate cancer in this locality. A prospective case study. Kenyatta National Referral Hospital and the Nairobi and Mater Hospitals. Fifty nine patients with advanced cancer of prostate (extra prostatic locally advanced and metastatic cancer). Transperineal trucut needle biopsies of the prostate revealed 15 patients (25.42%) had poorly differentiated cancers with Gleasons scores greater than 7. Fifteen patients (25.42%) had moderately differentiated cancers with Gleason scores of 6; and twenty nine other patients (49.2%) had well differentiated cancers with Gleason scores of 4 and below. Surgical castration was effected on 15 patients four of whom also had 50 mg of oral bicalutamide (casodex) daily. Thirty six patients were treated with subcutaneous goserelin (zoladex) depot 3.6 mg every 28 days. Ten of these patients also had 50 mg oral casodex daily in addition to the zoladex. Three patients in this group also had external radiotherapy for severe bone pains. Only eight patients were treated with oral diethylstilboestrol 3 mg daily. All the 15 patients with undifferentiated cancers died within 12 months. Of the 22 patients surviving at 48 months irrespective of the method of treatment, 20 of them had well differentiated cancers with Gleasons scores of 4 or less. Survival in the undifferentiated and poorly differentiated prostrate cancer Gleasons grades 4 and 5 with a score of over 7 is poor irrespective of the mode of treatment as all the patients in this group were dead within 12 months of diagnosis. Twenty patients (90.90%) of the surviving patients at 48 months had well differentiated cancers Gleasons grades 1 and 2 with scores of 4 or less indicating better prognosis for these tumours which are known to be slow growing with a much longer tumour doubling time.


Efficient gene targeting by homology-directed repair in rat zygotes using TALE nucleases.
The generation of genetically modified animals is important for both research and commercial purposes. The rat is an important model organism that until recently lacked efficient genetic engineering tools. Sequence-specific nucleases, such as ZFNs, TALE nucleases, and CRISPR/Cas9 have allowed the creation of rat knockout models. Genetic engineering by homology-directed repair (HDR) is utilized to create animals expressing transgenes in a controlled way and to introduce precise genetic modifications. We applied TALE nucleases and donor DNA microinjection into zygotes to generate HDR-modified rats with large new sequences introduced into three different loci with high efficiency (0.62%-5.13% of microinjected zygotes). Two of these loci (Rosa26 and Hprt1) are known to allow robust and reproducible transgene expression and were targeted for integration of a GFP expression cassette driven by the CAG promoter. GFP-expressing embryos and four Rosa26 GFP rat lines analyzed showed strong and widespread GFP expression in most cells of all analyzed tissues. The third targeted locus was Ighm, where we performed successful exon exchange of rat exon 2 for the human one. At all three loci we observed HDR only when using linear and not circular donor DNA. Mild hypothermic (30°C) culture of zygotes after microinjection increased HDR efficiency for some loci. Our study demonstrates that TALE nuclease and donor DNA microinjection into rat zygotes results in efficient and reproducible targeted donor integration by HDR. This allowed creation of genetically modified rats in a work-, cost-, and time-effective manner.


Accident at the Fukushima Dai-ichi nuclear power stations of TEPCO--outline & lessons learned.
The severe accident that broke out at Fukushima Dai-ichi nuclear power stations on March 11, 2011, caused seemingly infinite damage to the daily life of residents. Serious and wide-spread contamination of the environment occurred due to radioactive materials discharged from nuclear power stations (NPSs). At the same time, many issues were highlighted concerning countermeasures to severe nuclear accidents. The accident is outlined, and lessons learned are extracted with respect to the safety of NPSs, as well as radiation protection of residents under the emergency involving the accident. The materials of the current paper are those released by governmental agencies, academic societies, interim reports of committees under the government, and others.


Transcriptional regulation of the major histocompatibility complex (MHC) class I heavy chain, TAP1 and LMP2 genes by the human papillomavirus (HPV) type 6b, 16 and 18 E7 oncoproteins.
We have examined the possibility that the E7 proteins of the high-risk human papillomavirus (HPV) type 16 and 18 and the oncogenic adenovirus (Ad) type 12 E1A protein share the ability to down-regulate the expression of components of the antigen processing and presentation pathway, as a common strategy in the evasion of immune surveillance during the induction of cell transformation. Expression of the HPV 18 E7 oncoprotein, like Ad 12 E1A, resulted in repression of the major histocompatibility complex (MHC) class I heavy chain promoter, as well as repression of a bidirectional promoter that regulates expression of the genes encoding the transporter associated with antigen processing subunit 1 (TAP1) and a proteasome subunit, low molecular weight protein 2 (LMP2). HPV 16 E7 also caused a reduction in class I heavy chain promoter activity, however it did not have any significant effect on the activity of the bidirectional promoter. Interestingly, expression of the low-risk HPV 6b E7 protein resulted in an increase in MHC class I heavy chain promoter activity, while repressing the TAP1/LMP2 promoter. Interference with the class I pathway could also explain the ability of low-risk HPVs in inducing benign lesions.


[Comparison of several expressions of two-way ANOVA for a latent factor: analysis of the relation between music and mood].
The purpose of this research is to provide a two-way analysis of variance (ANOVA) model for a latent factor. Typical psychological studies measure mental states with questionnaires and analyze the variance of the measures into the portions attributable to various sources. This type of research, when conducted under regular ANOVA designs, uses total score as the dependent measures. However, this method is based on the unrealistic presumption that every item on the questionnaire has the same factor loading on the attribute being measured. In this research, we incorporated factor analysis model, and used a latent factor instead of total score as the dependent measure, thereby applying ANOVA under a more realistic assumption. Structural Equation Modeling (SEM) was used to express statistical models. This paper also examined a relation between music and mood, which is a quite popular area of research in psychology of music. To study the possible effects of tonality and key-signature on mood, music was chosen to represent tonality and key-signature conditions.


Current Clinical Applications and Future Potential of Diffusion Tensor Imaging in Traumatic Brain Injury.
In the setting of acute central nervous system (CNS) emergencies, computed tomography (CT) and conventional magnetic resonance imaging (MRI) play an important role in the identification of life-threatening intracranial injury. However, the full extent or even presence of brain damage frequently escapes detection by conventional CT and MRI. Advanced MRI techniques such as diffusion tensor imaging (DTI) are emerging as important adjuncts in the diagnosis of microstructural white matter injury in the acute and postacute brain-injured patient. Although DTI aids in detection of brain injury pathology, which has been repeatedly associated with typical adverse clinical outcomes, the evolution of acute changes and their long-term prognostic implications are less clear and the subject of much active research. A major aim of current research is to identify imaging-based biomarkers that can identify the subset of TBI patients who are at risk for adverse outcome and can therefore most benefit from ongoing care and rehabilitation as well as future therapeutic interventions.The aim of this study is to introduce the current methods used to obtain DTI in the clinical setting, describe a set of common interpretation strategies with their associated advantages and pitfalls, as well as illustrate the clinical utility of DTI through a set of specific patient scenarios. We conclude with a discussion of future potential for the management of TBI.


The hanta hunting study: underdiagnosis of Puumala hantavirus infections in symptomatic non-travelling leptospirosis-suspected patients in the Netherlands, in 2010 and April to November 2011.
Leptospirosis and haemorrhagic fever with renal syndrome (HFRS) are hard to distinguish clinically since these two important rodent-borne zoonoses share hallmark symptoms such as renal failure and haemorrhage. Leptospirosis is caused by infection with a spirochete while HFRS is the result of an infection with certain hantaviruses. Both diseases are relatively rare in the Netherlands. Increased incidence of HFRS has been observed since 2007 in countries that border the Netherlands. Since a similar rise in incidence has not been registered in the Netherlands, we hypothesise that due to overlapping clinical manifestations, hantavirus infections may be confused with leptospirosis, leading to underdiagnosis. Therefore, we tested a cohort of non-travelling Dutch patients with symptoms compatible with leptospirosis, but with a negative diagnosis, during 2010 and from April to November 2011. Sera were screened with pan-hantavirus IgG and IgM enzyme-linked immunosorbent assays (ELISAs). Sera with IgM reactivity were tested by immunofluorescence assay (IFA). ELISA (IgM positive) and IFA results were confirmed using focus reduction neutralisation tests (FRNTs). We found hantavirus-specific IgG and/or IgM antibodies in 4.3% (11/255) of samples taken in 2010 and in 4.1% (6/146) of the samples during the 2011 period. After FRNT confirmation, seven patients were classed as having acute Puumala virus infections. A review of hantavirus diagnostic requests revealed that at least three of the seven confirmed acute cases as well as seven probable acute cases of hantavirus infection were missed in the Netherlands during the study period.


Cell phone use and visual attention.
The purpose of the present study was to evaluate how much cell phones and just speaking (similar to speaking to someone in the car vs a hands-free cell phone task) interfere with visual attention skills as might be required in a driving situation. Influence of cell phones on attention has been noted but little research has been completed. Licensed adult drivers were divided into three groups (ns = 15) with all subjects taking the Connors Continuous Performance Test II. Group 1 performed without any distractions: those in Group 2 performed with someone in the same room talking to them: Group 3 engaged in a cell phone conversation during the task. Overall, there were substantial differences among groups on all variables, but primarily between the control group and the two experimental groups. While the cell phone group had lower mean scores than the talking group overall, the differences were not significant. Thus, while cell phones were distracting to visual attention functions on the Connors task, they were not more distracting than a similarly active conversation without a cell phone.


Community-associated meticillin-resistant Staphylococcus aureus as a cause of hospital-acquired infections.
The worldwide emergence of community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA) during the last decade represents a significant change in the biology of MRSA strains and is changing the epidemiology of MRSA infections. CA-MRSA infections are caused by strains belonging to lineages distinct from HA-MRSA. In the community, CA-MRSA strains typically cause skin and soft tissue infections in children and younger adults. However, CA-MRSA strains increasingly cause healthcare-acquired infections including surgical site infections, ventilator-associated pneumonia and bacteraemia. A mathematical model showing the influence of MRSA transmission in the community on the prevalence of MRSA in hospitals is presented. The increasing prevalence of MRSA in the community also results in an increase in community-onset MRSA (CO-MRSA) among S. aureus bacteraemia and other invasive infections. These patients do not have typical risk factors for MRSA. Such changes may have profound implications for the choice of empirical therapy for serious infections where S. aureus is a possible cause. The new and potentially very large reservoir of MRSA in production animals with subsequent transmission to humans represents an additional serious threat to the control of MRSA both in general and as a cause of healthcare-acquired infections. CA-MRSA is thus a matter of serious concern and should be suppressed.


The outcome of unretrieved gallstones in the peritoneal cavity during laparoscopic cholecystectomy. A prospective analysis.
Gallbladder perforation during laparoscopic cholecystectomy (LC) with spillage of bile and gallstones occurs in a substantial number of patients (up to 40%). Most surgeons believe that free intraperitoneal stones are not a justification for conversion to laparotomy even if a large number of stones are left in situ. There are, however, a number of reports demonstrating that, on occasion, these unretrieved gallstones may cause infection or abscess, inflammation, fibrosis, adhesions, cutaneous sinuses, small bowel obstruction, or generalized septicemia. The aim of this study was to determine the outcome of unretrieved gallstones in the peritoneal cavity after gallbladder perforation during LC. In a 7-year period between 1989 and 1996, prospective data were maintained on 856 patients who underwent LCs by a single surgeon (R. J.F.). Of the 856 patients, 165 (16%) had gallbladder perforations resulting in lost gallstones in the peritoneal cavity. A concerted attempt was made to remove the lost stones using a variety of extraction devices. Of these 165 patients, 106 (64%) were available for follow-up through mail (76%) and by telephone (24%). The mean age of these patients was 64.9 years (range, 18 to 98 years), and the mean follow-up was 44.8 months (range 4.9 to 92.3 months). Of the 106 patients with unretrieved gallstones, we identified four patients with short-term complications and one patient with a long-term complication. The first patient with a short-term complication had pyrexia for 10 days postoperatively. Diagnostic evaluation, which included computed tomography (CT) scan, failed to reveal any abnormality. The patient was treated conservatively with a course of oral antibiotics. In the second patient, cellulitis developed at a drain site after its removal, which resolved with oral antibiotics. The third patient acquired an umbilical wound abscess, which drained spontaneously, requiring no treatment. A sterile subphrenic collection developed in the fourth patient 1 month postoperatively, which was treated with percutaneous drainage under CT guidance. The only long-term complication was spontaneous erosion of a gallstone from the back of a patient with a questionable history of inflammatory bowel disease 8 months postoperatively. All of the patients made complete recoveries. In most patients, unretrieved gallstones are of no consequence, but complications occur occasionally. It is therefore advisable to retrieve as many gallstones as possible during LC short of converting to a laparotomy.


Improved Accuracy on Lateralized Spatial Judgments in Healthy Aging.
Healthy young adults often demonstrate a leftward spatial bias called "pseudoneglect" which often diminishes with aging. One hypothesis for this phenomenon is an age-related deterioration in right hemisphere functions (right hemi-aging). If true, then a greater rightward bias should be evident on all spatial attention tasks regardless of content. Another hypothesis is a decrease in asymmetrical hemispheric activation with age (HAROLD). If true, older participants may show reduced bias in all spatial tasks, regardless of leftward or rightward biasing of specific spatial content. Seventy right-handed healthy participants, 33 younger (21-40) and 37 older (60-78), were asked to bisect solid and character-letter lines as well as to perform left and right trisections of solid lines. Both groups deviated toward the left on solid line bisections and left trisections. Both groups deviated toward the right on right trisections and character line bisections. In all tasks, the older participants were more accurate than the younger participants. The finding that older participants were more accurate than younger participants across all bisection and trisection conditions suggests a decrease in the asymmetrical hemispheric activation of these specialized networks important in the allocation of contralateral spatial attention or spatial action intention.


The crystallization of ultralong normal paraffins: the onset of chain folding.
The nature of chain folding in polymers and the determination of the chain length at which folding occurs have been central questions in polymer science. The study of the formation of lamellar polymer crystals through chain folding has received a new impetus as a result of the recent synthesis of normal alkanes of strictly uniform chain lengths up to C(390) H(782). Chain folding is found in all such paraffins starting with C(150)H(302). As with polyethylenes obtained by conventional polymerization, the fold length in the normal alkanes varies with crystallization temperature, but it is always an integral reciprocal of the full chain length. This behavior indicates that the methyl end groups are located at the lamellar surface and that the fold itself must be sharp and adjacently reentrant.


HCN4 subunit expression in fast-spiking interneurons of the rat spinal cord and hippocampus.
Hyperpolarisation-activated (Ih) currents are considered important for dendritic integration, synaptic transmission, setting membrane potential and rhythmic action potential (AP) discharge in neurons of the central nervous system. Hyperpolarisation-activated cyclic nucleotide-gated (HCN) channels underlie these currents and are composed of homo- and hetero-tetramers of HCN channel subunits (HCN1-4), which confer distinct biophysical properties on the channel. Despite understanding the structure-function relationships of HCN channels with different subunit stoichiometry, our knowledge of their expression in defined neuronal populations remains limited. Recently, we have shown that HCN subunit expression is a feature of a specific population of dorsal horn interneurons that exhibit high-frequency AP discharge. Here we expand on this observation and use neuroanatomical markers to first identify well-characterised neuronal populations in the lumbar spinal cord and hippocampus and subsequently determine whether HCN4 expression correlates with high-frequency AP discharge in these populations. In the spinal cord, HCN4 is expressed in several putative inhibitory interneuron populations including parvalbumin (PV)-expressing islet cells (84.1%; SD: ±2.87), in addition to all putative Renshaw cells and Ia inhibitory interneurons. Similarly, virtually all PV-expressing cells in the hippocampal CA1 subfield (93.5%; ±3.40) and the dentate gyrus (90.9%; ±6.38) also express HCN4. This HCN4 expression profile in inhibitory interneurons mirrors both the prevalence of Ih sub-threshold currents and high-frequency AP discharge. Our findings indicate that HCN4 subunits are expressed in several populations of spinal and hippocampal interneurons, which are known to express both Ih sub-threshold currents and exhibit high-frequency AP discharge. As HCN channel function plays a critical role in pain perception, learning and memory, and sleep as well as the pathogenesis of several neurological diseases, these findings provide important insights into the identity and neurochemical status of cells that could underlie such conditions.


Personality and individual difference correlates of positive body image.
In the present study, 101 women and 106 men from a community sample of British adults completed the Body Appreciation Scale (BAS), along with a battery of individual difference measures and demographics. Contrary to previous findings, there were no sex differences in BAS scores, either before or after controlling for individual differences in other measures. The results also showed that, moderating for participants' sex, self-assessed attractiveness, educational qualifications, neuroticism, extraversion, and BMI were all significant predictors of body appreciation. In addition, higher media consumption and higher (male-stereotypic) instrumentality were associated with, but did not predict, higher body appreciation. These results are discussed in relation to the extant work on body image.


Thromboprophylaxis in non-surgical patients: who, when and how?
Because of the serious lack of useable, relevant information, most recommendations for prevention of thrombosis in non-surgical patients are extrapolations from much larger clinical trials experienced in surgery. Directly relevant evidence comes predominantly from very small randomized trials, many of them open label and carried out more than 20 years before the introduction of more recent and important changes in clinical care that may have substantially reduced the baseline thrombosis risk. In these early studies, low-dose heparin and low-molecular-weight heparins prevented subclinical deep vein thrombosis in ischaemic stroke, myocardial infarction and among elderly medical inpatients. Although it is likely that these drugs also prevent subclinical deep vein thrombosis after spinal cord injury or other major trauma, and when patients require intensive medical care, the supporting evidence in these circumstances comes mainly from cohort studies and poorly controlled comparisons. In contrast, the heparins have not reduced mortality or demonstrably prevented pulmonary embolism after ischaemic stroke or among elderly medical inpatients in large and well-conducted clinical endpoint trials, from which no clinically important benefit could be demonstrated. From analyses it is suggested that such benefit is probably more difficult to demonstrate for medical than for surgical patients. In the absence of sufficient information that is specific to medical patients, various forms of prophylaxis known to be effective in surgery will continue to be applied in high-risk individuals. After venous thromboembolism, it now appears that the best duration of oral anticoagulant therapy to prevent a recurrence is determined to a greater extent by whether the thrombotic episode was idiopathic or triggered by a clinically recognizable cause, whether it was transient or continuing, and whether the deep vein thrombosis was extensive, limited to the calf veins or was a first or recurrent event.


Bioassay of 3-amino-9-ethylcarbazole hydrochloride for possible carcinogenicity.
A bioassay of 3-amino-9-ethylcarbazole (hydrochloride) for possible carcinogenicity was conducted by administering the test chemical in feed to Fischer 344 rats and B6C3F1 mice. Both the free amine and the hydrochloride salt were used. Groups of 50 rats of each sex and 50 mice of each sex were administered the test chemical at one of two doses, either 800 or 2,000 ppm for rats and either 800 or 1,200 ppm for mice, for 78 weeks. The rats were then observed for an additional 26-29 weeks, and the mice for an additional 16-17 weeks. Controls consisted of groups of 50 untreated rats of each sex and 50 untreated mice of each sex; separate controls were used for the groups of animals administered the different doses. All surviving rats were killed at 104-110 weeks; all surviving mice were killed at 94-97 weeks. Since the suppliers of the low-dose rats and mice differed from those of the corresponding low-dose controls, while the suppliers for the high-dose rats and mice were the same as those of the corresponding high-dose controls, comparisons of the high-dose groups with their corresponding controls were the most appropriate. Furthermore, since the low-dose animals did not receive the same regimen of administration of the test compound as that received by the high-dose animals, and since tests using the low-dose groups were not performed concurrently with those using the high-dose groups, analyses of the dose-related trends were not possible. Although the interpretation of results of the study was based primarily on comparisons of high-dose groups with their respective controls, the results obtained with the low-dose groups, regardless of the indicated complicating factors, supported the interpretation. Neoplasms of the liver were observed in significant incidences in rats and mice of both sexes. In male rats, hepatocellular carcinomas alone were significantly higher (P</=0.020) in both the low- and high-dose groups. When neoplastic nodules of the liver were combined with hepatocellular carcinomas, the combination occurred at significant incidences (P</= 0.012) in the low- and high-dose male rats and in the high-dose female rats (males: low-dose controls 0/36, low-dose 12/42; high-dose controls 1/48, high-dose 22/48; females: high-dose controls 0/50, high-dose 6/48). Hepatocellular carcinomas alone similarly occurred at significant incidences (P<0.001) in the low- and high-dose male and female mice (males: low-dose controls 7/48, low-dose 32/44; high-dose controls 6/44, high-dose 41/49; females: low-dose controls 1/47, low-dose 36/43; high-dose controls 1/45; high-dose 43/49). Papillomas or carcinomas of the integumentary system occurred at significant incidences (P</= 0.013) in both the low- and high-dose male rats (low-dose controls 0/36, low-dose 8/44; high-dose controls 0/48, high-dose 6/48); these tumors also occurred in low- and high-dose female rats, but not at incidences high enough to be statistically significant (low-dose control 0/39, low-dose 4/44; high-dose controls 0/50, high-dose 4/49). Carcinomas of the Zymbal's glands of the ear occurred at significant incidences (P</= 0.045) in the low- and high-dose male and female rats (males: low-dose controls 0/36, low-dose 5/44; high-dose controls 0/48, high-dose 7/48; females: low-dose controls 0/39, low-dose 10/44; high-dose controls 0/50, high-dose 12/49). These tumors were first observed as early as week 47 on study. Adenocarcinomas of the uterus or endometrium occurred at a significant incidence (P=0.002) only in the high-dose female rats (high-dose controls 1/50, high-dose 11/49); these tumors also occurred in the low-dose females, but not at an incidence high enough to be statistically significant (low-dose controls 4/38, low dose 11/43). It is concluded that under the conditions of this bioassay, 3-amino-9-ethylcarbazole (hydrochloride) was carcinogenic for the liver, inducing hepatocellular carcinomas in Fischer 344 rats and B6C3F1 mice of both sexes. Other tumors induced in the rats were carcinomas or papillomas of the integumentary system in males, carcinomas of the Zymbal's gland of the ear in males and females, and adenocarcinomas of the uterus.


How to use unfractionated heparin to treat neonatal thrombosis in clinical practice.
Among children, neonates have the highest incidence of thrombosis due to risk factors such as catheter instrumentation, an evolving coagulation system and congenital heart disease. Unfractionated heparin (UFH) is one of the most commonly used anticoagulants in neonates. Published guidelines delineate dosing and monitoring protocols for UFH therapy in newborns. However, challenging clinical situations frequently present that warrants healthcare providers to think critically beyond the range of guidelines, and judiciously resolve specific problems. This review focuses briefly on the epidemiology of neonatal thrombosis and the use of UFH in this population. It is followed by a discussion on dosing of UFH in neonates, limited evidence that forms the basis of published guidelines with justification for a treatment regimen that precludes the use of a heparin loading dose in newborns and monitoring of UFH therapy with currently available tests such as antifactor Xa (anti-Xa) level and activated partial thromboplastin time (APTT). Multiple studies have demonstrated a lack of correlation between anti-Xa levels and APTT as well as between different anti-Xa assays. Many centers world-wide rely only on APTT for monitoring purposes and do not have access to anti-Xa assays. To address these difficulties, we propose two practical algorithms, with and without the use of anti-Xa levels that clinicians can follow when monitoring UFH therapy in neonates. The article concludes with an overview of the side-effects of UFH.


Resolution of spatial and temporal visual attention in infants with fragile X syndrome.
Fragile X syndrome is the most common cause of inherited intellectual impairment and the most common single-gene cause of autism. Individuals with fragile X syndrome present with a neurobehavioural phenotype that includes selective deficits in spatiotemporal visual perception associated with neural processing in frontal-parietal networks of the brain. The goal of the current study was to examine whether reduced resolution of spatial and/or temporal visual attention may underlie perceptual deficits related to fragile X syndrome. Eye tracking was used to psychophysically measure the limits of spatial and temporal attention in infants with fragile X syndrome and age-matched neurotypically developing infants. Results from these experiments revealed that infants with fragile X syndrome experience drastically reduced resolution of temporal attention in a genetic dose-sensitive manner, but have a spatial resolution of attention that is not impaired. Coarse temporal attention could have significant knock-on effects for the development of perceptual, cognitive and motor abilities in individuals with the disorder.


Mid-infrared 2000-nm bandwidth supercontinuum generation in suspended-core microstructured sulfide and tellurite optical fibers.
In this work, we report the experimental observation of supercontinua generation in two kinds of suspended-core microstructured soft-glass optical fibers. Low loss, highly nonlinear, tellurite and As2S3 chalcogenide fibers have been fabricated and pumped close to their zero-dispersion wavelength in the femtosecond regime by means of an optical parametric oscillator pumped by a Ti:Sapphire laser. When coupled into the fibers, the femtosecond pulses result in 2000-nm bandwidth supercontinua reaching the Mid-Infrared region and extending from 750 nm to 2.8 µm in tellurite fibers and 1 µm to 3.2 µm in chalcogenide fibers, respectively.


HIV and aging: insights from the Asia Pacific HIV Observational Database (APHOD).
The proportion of people living with HIV/AIDS in the ageing population (>50 years old) is increasing. We aimed to explore the relationship between older age and treatment outcomes in HIV-positive persons from the Asia Pacific region. Patients from the Australian HIV Observational Database (AHOD) and the TREAT Asia HIV Observational Database (TAHOD) were included in the analysis. We used survival methods to assess the association between older age and all-cause mortality, as well as time to treatment modification. We used regression analyses to evaluate changes in CD4 counts after combination antiretroviral therapy (cART) initiation and determined the odds of detectable viral load, up to 24 months of treatment. A total of 7142 patients were included in these analyses (60% in TAHOD and 40% in AHOD), of whom 25% were >50 years old. In multivariable analyses, those aged > 50 years were at least twice as likely to die as those aged 30-39 years [hazard ratio (HR) for 50-59 years: 2.27; 95% confidence interval (CI) 1.34-3.83; HR for > 60 years: 4.28; 95% CI 2.42-7.55]. The effect of older age on CD4 count changes was insignificant (p-trend=0.06). The odds of detectable viral load after cART initiation decreased with age (p-trend=< 0.0001). The effect of older age on time to first treatment modification was insignificant (p-trend=0.21). We found no statistically significant differences in outcomes between AHOD and TAHOD participants for all endpoints examined. The associations between older age and typical patient outcomes in HIV-positive patients from the Asia Pacific region are similar in AHOD and TAHOD. Our data indicate that 'age effects' traverse the resource-rich and resource-limited divide and that future ageing-related findings might be applicable to each setting.


Tyrosine phosphorylation of p62dok by p210bcr-abl inhibits RasGAP activity.
The t(9;22) chromosomal translocation is found in almost all patients with chronic myelogenous leukemia. The resultant Bcr-Abl fusion gene expresses a chimeric fusion protein p210(bcr-abl) with increased tyrosine kinase activity. Hematopoietic progenitors isolated from chronic myelogenous leukemia patients in the chronic phase contain constitutively tyrosine-phosphorylated p62(dok) protein. p62(dok) associates with the Ras GTPase-activating protein (RasGAP), but only when p62(dok) is tyrosine phosphorylated. Here we have investigated the interaction between p62(dok) and RasGAP and the consequences of p62(dok) tyrosine phosphorylation on the activity of RasGAP. We have found that p62(dok) is directly tyrosine phosphorylated by p210(bcr-abl), and the sites of phosphorylation are located in the C-terminal half of the p62(dok) molecule. We have identified five tyrosine residues that are involved in in vitro RasGAP binding and have found that tyrosine-phosphorylated p62(dok) inhibits RasGAP activity. Our results suggest that p210(bcr-abl) might lead to the activation of the Ras signaling pathway by inhibiting a key down-regulator of Ras signaling.


High-resolution secondary ion mass spectrometry reveals the contrasting subcellular distribution of arsenic and silicon in rice roots.
Rice (Oryza sativa) takes up arsenite mainly through the silicic acid transport pathway. Understanding the uptake and sequestration of arsenic (As) into the rice plant is important for developing strategies to reduce As concentration in rice grain. In this study, the cellular and subcellular distributions of As and silicon (Si) in rice roots were investigated using high-pressure freezing, high-resolution secondary ion mass spectrometry, and transmission electron microscopy. Rice plants, both the lsi2 mutant lacking the Si/arsenite efflux transporter Lsi2 and its wild-type cultivar, with or without an iron plaque, were treated with arsenate or arsenite. The formation of iron plaque on the root surface resulted in strong accumulation of As and phosphorous on the epidermis. The lsi2 mutant showed stronger As accumulation in the endodermal vacuoles, where the Lsi2 transporter is located in the plasma membranes, than the wild-type line. As also accumulated in the vacuoles of some xylem parenchyma cells and in some pericycle cells, particularly in the wild-type mature root zone. Vacuolar accumulation of As is associated with sulfur, suggesting that As may be stored as arsenite-phytochelatin complexes. Si was localized in the cell walls of the endodermal cells with little apparent effect of the Lsi2 mutation on its distribution. This study reveals the vacuolar sequestration of As in rice roots and contrasting patterns of As and Si subcellular localization, despite both being transported across the plasma membranes by the same transporters.


Contribution of SAM and HD domains to retroviral restriction mediated by human SAMHD1.
The human SAMHD1 protein is a novel retroviral restriction factor expressed in myeloid cells. Previous work has correlated the deoxynucleotide triphosphohydrolase activity of SAMHD1 with its ability to block HIV-1 and SIV(mac) infection. SAMHD1 is comprised of the sterile alpha motif (SAM) and histidine-aspartic (HD) domains; however the contribution of these domains to retroviral restriction is not understood. Mutagenesis and deletion studies revealed that expression of the sole HD domain of SAMHD1 is sufficient to achieve potent restriction of HIV-1 and SIV(mac). We demonstrated that the HD domain of SAMHD1 is essential for the ability of SAMHD1 to oligomerize by using a biochemical assay. In agreement with previous observations, we mapped the RNA-binding ability of SAMHD1 to the HD domain. We also demonstrated a direct interaction of SAMHD1 with RNA by using enzymatically-active purified SAMHD1 protein from insect cells. Interestingly, we showed that double-stranded RNA inhibits the enzymatic activity of SAMHD1 in vitro suggesting the possibility that RNA from a pathogen might modulate the enzymatic activity of SAMHD1 in cells. By contrast, we found that the SAM domain is dispensable for retroviral restriction, oligomerization and RNA binding. Finally we tested the ability of SAMHD1 to block the infection of retroviruses other than HIV-1 and SIV(mac). These results showed that SAMHD1 blocks infection of HIV-2, feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV), Equine infectious anemia virus (EIAV), N-tropic murine leukemia virus (N-MLV), and B-tropic murine leukemia virus (B-MLV).


Prognostic value of self-reported fatigue on overall survival in patients with myelodysplastic syndromes: a multicentre, prospective, observational, cohort study.
The clinical presentation of myelodysplastic syndromes is highly variable and so accurate prediction of outcomes in these patients is crucial. We aimed to assess whether self-reported fatigue severity predicts overall survival beyond gold-standard prognostic indices in patients with higher-risk myelodysplastic syndromes. We did a multicentre, prospective, observational, cohort study of patients from 37 centres in Europe, USA, and east Asia. Adults (≥18 years) with myelodysplastic syndromes were consecutively enrolled within 6 months of diagnosis with an intermediate-2-risk or high-risk score according to the International Prognostic Scoring System (IPSS). Patients were enrolled irrespective of older age, comorbidities, performance status, and progression from a lower IPSS risk score category. All patients had to complete a quality of life assessment at baseline. With use of univariate and then multivariate Cox proportional hazards regression analysis, we constructed a multivariate model of how prognostic variables, including IPSS and fatigue score from the European Organisation for Research and Treatment of Cancer quality-of-life questionnaire-core 30, predicted overall survival. The primary endpoint was overall survival by baseline self-reported fatigue scale ratings. This study was registered with ClinicalTrials.gov, number NCT00809575. Between Nov 10, 2008, and Aug 13, 2012, we enrolled 280 patients with a median age of 71 years (IQR 64-77). The median follow-up was 15 months (IQR 8-27), and the last patient was assessed Feb 16, 2015. The median overall survival from diagnosis was 17 months (95% CI 15-19). In univariate analysis, the baseline factors that were significantly associated with reduced overall survival were increasing age, transfusion dependency (defined as having received at least one red blood cell transfusion every 8 weeks over a period of 4 months), Eastern Cooperative Oncology Group (ECOG) performance status of two or more, increased white blood cell count, high-risk IPSS score, and higher self-reported fatigue severity. In multivariate analysis, baseline factors independently associated with reduced overall survival were high-risk IPSS score (hazard ratio [HR] 2·525, 95% CI 1·357-4·697; p=0·0035) and a higher score for fatigue (1·110, 1·040-1·170, for every ten points of fatigue deterioration; p=0·0007). In further multivariate models for survival, including either the WHO-based prognostic scoring system or the revised version of the IPSS classification, fatigue remained a statistically significant independent prognostic factor with a HR of 1·120 (1·050-1·180, p=0.0003) and a HR of 1·130 (1·060-1·190, p=0·0002), respectively. In patients with newly diagnosed higher-risk myelodysplastic syndromes, self-reported fatigue severity provides prognostic information for survival independent from gold-standard risk classifications. Our findings suggest that fatigue assessment should be included in routine diagnostic investigation for these patients and considered as a standard baseline stratification factor in future randomised controlled trials. Associazione Italiana contro le Leucemie, Linfomi e Mieloma (AIL).


Adolescents Who Self-Harm: The Patterns in Their Interpersonal and Psychosocial Difficulties.
We proposed that having mutually hostile interactions with others is a strong environmental stress factor that, together with diverse psychosocial problems, characterizes adolescents who self-harm. Using cluster analysis, this study examined the naturally occurring patterns of hostility conditions and psychosocial difficulties in a normative sample of 2,029 adolescents (50% boys; Mage = 13.89). Results showed that self-harming behavior was significantly higher among the subgroup of adolescents with mutually hostile interactions who exhibited both internalizing and externalizing problems than among adolescents with other interpersonal-psychosocial configurations. Also, this subgroup of adolescents reported high impulsivity, anger dysregulation, and low self-esteem. These findings support recent research that indicates that adolescents who self-harm also tend to expose others to hostility and display externalizing symptoms.


Human MAIT cells are rapidly activated by Aspergillus spp. in an APC-dependent manner.
Mucosal associated invariant T cells (MAIT cells) are innate-like T cells (TC) which are known to be activated by several bacteria and viruses. However, activation of MAIT cells by moulds, such as the opportunistic human pathogen Aspergillus, is not well described. Stimulation of human PBMC with A. fumigatus, A. flavus, or A. terreus conidia revealed that in contrast to conventional CD4+ and CD8+ TC, MAIT cells responded already after 4 h of coincubation with upregulation of CD69. Furthermore, concurrent increase of CD107a expression and reduced intracellular expression of cytolytic proteins like perforin and granzyme indicated degranulation of intracellular vesicles. MAIT cell activation only occurred in the presence of APC and was dependent on cell-cell contact as separation of TC and APC abrogated MAIT cell activation. Furthermore, we observed that MAIT cell activation by moulds requires presentation of riboflavin metabolites and depends on TCR engagement as antibody blocking of MR1, the antigen presenting molecule for MAIT cells, prevented upregulation of CD69 and CD107a. In summary, we could demonstrate that MAIT cells are activated by Aspergillus conidia in a TCR-dependent manner by APC. These findings reveal MAIT cells as an interesting new target in antifungal defense.


Enhanced plant nutrient use efficiency with PGPR and AMF in an integrated nutrient management system.
A 3 year field study was conducted with field corn from 2005 to 2007 to test the hypothesis that microbial inoculants that increase plant growth and yield can enhance nutrient uptake, and thereby remove more nutrients, especially N, P, and K from the field as part of an integrated nutrient management system. The field trial evaluated microbial inoculants, which include a commercially available plant growth-promoting rhizobacteria (PGPR), arbuscular mycorrhiza fungi (AMF), and their combination across 2 tillage systems (no-till and conventional till) and 2 fertilization regimes (poultry litter and ammonium nitrate). Data were collected on plant height, yield (dry mass of ears and silage), and nutrient content of corn grain and silage. In addition, nutrient content of soil was determined, and bioavailability of soil nutrient was measured with plant root simulator probes. Results showed that inoculants promoted plant growth and yield. For example, grain yields (kg.ha(-1)) in 2007 for inoculants were 7717 for AMF, 7260 for PGPR+AMF, 7313 for PGPR, 5725 for the control group, and for fertilizer were 7470 for poultry litter and 6537 for NH4NO3. Nitrogen content per gram of grain tissues was significantly enhanced in 2006 by inoculant, fertilizer, and their interactions. Significantly higher amounts of N, P, and K were removed from the plots with inoculants, based on total nutrient content of grain per plot. These results supported the overall hypothesis and indicate that application of inoculants can lead to reduction in the build up of N, P, and K in agricultural soils. Further studies should be conducted to combine microbial inoculants with reduced rates of fertilizer.


mTOR complex 1 regulates lipin 1 localization to control the SREBP pathway.
The nutrient- and growth factor-responsive kinase mTOR complex 1 (mTORC1) regulates many processes that control growth, including protein synthesis, autophagy, and lipogenesis. Through unknown mechanisms, mTORC1 promotes the function of SREBP, a master regulator of lipo- and sterolgenic gene transcription. Here, we demonstrate that mTORC1 regulates SREBP by controlling the nuclear entry of lipin 1, a phosphatidic acid phosphatase. Dephosphorylated, nuclear, catalytically active lipin 1 promotes nuclear remodeling and mediates the effects of mTORC1 on SREBP target gene, SREBP promoter activity, and nuclear SREBP protein abundance. Inhibition of mTORC1 in the liver significantly impairs SREBP function and makes mice resistant, in a lipin 1-dependent fashion, to the hepatic steatosis and hypercholesterolemia induced by a high-fat and -cholesterol diet. These findings establish lipin 1 as a key component of the mTORC1-SREBP pathway.


A novel transgenic mouse model of lysosomal storage disorder.
Knockout technology has proven useful for delineating functional roles of specific genes. Here we describe and provide an explanation for striking pathology that occurs in a subset of genetically engineered mice expressing a rat CaVβ2a transgene under control of the cardiac α-myosin heavy chain promoter. Lesions were limited to mice homozygous for transgene and independent of native Cacnb2 genomic copy number. Gross findings included an atrophied pancreas; decreased adipose tissue; thickened, orange intestines; and enlarged liver, spleen, and abdominal lymph nodes. Immune cell infiltration and cell engulfment by macrophages were associated with loss of pancreatic acinar cells. Foamy macrophages diffusely infiltrated the small intestine's lamina propria, while similar macrophage aggregates packed liver and splenic red pulp sinusoids. Periodic acid-Schiff-positive, diastase-resistant, iron-negative, Oil Red O-positive, and autofluorescent cytoplasm was indicative of a lipid storage disorder. Electron microscopic analysis revealed liver sinusoids distended by clusters of macrophages containing intracellular myelin "swirls" and hepatocytes with enlarged lysosomes. Additionally, build up of cholesterol, cholesterol esters, and triglycerides, along with changes in liver metabolic enzyme levels, were consistent with a lipid processing defect. Because of this complex pathology, we examined the transgene insertion site. Multiple transgene copies inserted into chromosome 19; at this same site, an approximate 180,000 base pair deletion occurred, ablating cholesterol 25-hydroxylase and partially deleting lysosomal acid lipase and CD95 Loss of gene function can account for the altered lipid processing, along with hypertrophy of the immune system, which define this phenotype, and serendipitously provides a novel mouse model of lysosomal storage disorder.


[Quality standard study on Tibetan medicine Gentianae Szechenyii Flos].
In order to efficiently control the quality of the Tibetan medicine Gentianae Szechenyii Flos, the quality standard was established in this study. The tests of water content, total ash and ethanol-soluble extractives of the crude drugs were carried out based on the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC method was established by using reference drug and gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid (7: 1.5: 1: 0.2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on a Ultimate XB-C18 (4.6 mm x 250 mm, 5 μm) column, using methanol-water (0.02% phosphoric acid) (52:48) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is 25 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 10.01-400.32 mg x L(-1) with the regression equation of Y = 1 539.5X - 33.339 (r = 0.999 7), and the average recovery was 99.68% (RSD 1.92%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 19 batches samples were varied in the ranges of 14.48-31.51 mg x g(-1), 11.25% -12.74% and 24.21% - 31.60%, respectively, and total ash was 4.64% - 6.12% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 15.0 mg x g(-1) (1.5%) and 21.0%, respectively; the water and total ash are not more than 13.0% and 6.0%, respectively.


Recent Progress on Chemical Biology of Pluripotent Stem Cell Self-renewal, Reprogramming and Cardiomyogenesis.
Pluripotent stem cells, such as embryonic stem (ES) cells and induced pluripotent stem (iPS) cells, hold great promise as a cell source for regenerative therapies to treat many major diseases characterized by an irreversible loss of functional tissues. However, the future clinical application of pluripotent stem cells faces a number of obstacles regarding the safety, efficiency and long-term benefits. Some of these challenges are being addressed by the chemical biology approach using small molecules. In this paper, we review the recent progress and patents on small molecules which promote pluripotent stem cell maintenance, reprogramming, and direct differentiation with a focus on cardiomyogenesis.


Evaluating the effectiveness of a patient-oriented hand rehabilitation programme.
This study evaluated the effectiveness of a patient-oriented, hand rehabilitation programme compared to a standard programme regarding functional outcomes, return to work, patient satisfaction and costs. Patients were recruited in two consecutive cohorts. One cohort received the standard treatment programme (n = 75) and the other a programme based on principles of patient orientation (n = 75). Data were collected at the beginning and end of rehabilitation and 6 months after discharge. Clinical variables included range of motion, grip and pinch strength. Self-reported measures included pain, upper extremity functioning, health status, satisfaction and job situation. Analysis of variance for repeated measurements was used to calculate the main effects. The patient-oriented group showed more favourable results with respect to DASH scores (P <.05), pain (P <.001) and patient satisfaction (P <.0001). More patients returned to their former jobs and time off sick was reduced. We concluded that the patient-oriented approach was more effective and cost-saving.


Neutralization of coral snake Micrurus nigrocinctus venom by a monovalent antivenom.
1. The neutralizing ability of a monovalent anti-Micrurus nigrocinctus (coral snake) antivenom produced in Costa Rica was tested against the lethal, myotoxic and phospholipase A2 activities of homologous venom. In addition, immunodiffusion and Western blot analyses were performed. 2. In experiments where venom and antivenom were incubated prior to the test, antivenom was effective in neutralizing lethal, myotoxic and phospholipase A2 activities, with Effective Doses 50% of 2700 microliters antivenom/mg venom, 1840 microliters antivenom/mg venom, and 3630 microliters antivenom/mg venom, respectively. 3. When coral snake antivenom was administered at different times after coral snake venom injection, neutralization of lethality was achieved when antivenom was injected i.v. immediately and 15 min after venom. In contrast, lethality was not reduced when antivenom was administered by the im route. Only partial neutralization of myotoxicity was observed even when antivenom was injected i.v. immediately after envenomation. 4. Immunodiffusion and immunoblot analyses demonstrated the presence of antibodies in antivenom against several, but not all, venom components.


Identification and characterization of vascular endothelial growth factor receptor (Flt) in bovine retinal pericytes.
Vascular endothelial growth factor (VEGF) plays an important role in the hypoxia-stimulated neovascularization of ischemic retinal diseases such as proliferative diabetic retinopathy. VEGF exerts its effect through two known high-affinity tyrosine kinase receptors, named kinase insert domain-containing receptor (KDR) and the fms-like tyrosine kinase (Flt). VEGF receptors are located primarily on endothelial cells, although receptors on a few other nonocular cell types also have been described. In the present study, we demonstrate the expression of Flt, but not KDR, in bovine retinal pericytes (BRPCs). Although KDR is expressed predominantly in retinal endothelial cells, Northern blot analysis demonstrated substantial expression of the Flt gene in BRPCs without detection of KDR despite using polyadenylated RNA. Hypoxia increased Flt gene expression in BRPCs (2.7-fold, P < 0.01). 125I-labeled VEGF binding analysis on BRPCs demonstrated two apparent high-affinity receptor subtypes (Kd = 14 and 215 pmol/l), with 2.9 x 10(4) and 1.4 x 10(5) receptors/cell, respectively. 125I-VEGF affinity cross-linking demonstrated VEGF-specific binding complexes at 150, 172, 187, and 200 kDa under reducing conditions. Western blot analysis using an anti-phosphotyrosine antibody demonstrated VEGF-induced tyrosine phosphorylation of several proteins. VEGF stimulation had little effect on initial BRPCs growth rates but significantly increased BRPCs number after 7 days. These results suggest that two classes of high-affinity VEGF receptors are present on BRPCs, at least one of which is analogous to Flt and is capable of intracellular protein phosphorylation. Thus, VEGF might regulate the function of both retinal endothelial cells and retinal pericytes to induce pathological angiogenesis and vascular remodeling during proliferative diabetic retinopathy and other ischemic retinal diseases.


Feasibility and safety of a 6-month exercise program to increase bone and muscle strength in children with juvenile idiopathic arthritis.
Arthritis in childhood can be associated with muscle weakness around affected joints, low bone mass and low bone strength. Exercise is recognized as an important part of management of children with juvenile idiopathic arthritis (JIA) but the exercise prescription to best promote bone and muscle health is unknown. We therefore aimed to: 1. assess feasibility and safety of a 6-month home- and group-based exercise program for children with JIA; 2. estimate the effect of program participation on bone mass and strength, muscle function and clinical outcomes and 3. determine if any positive changes in bone and muscle outcomes are maintained 6 months later. We recruited 24 children with JIA who were part of the Linking Exercise, Physical Activity and Pathophysiology in Childhood Arthritis (LEAP) study to participate in a 6-month home-based exercise program involving jumping and handgrip exercises, resistance training and one group exercise session per month. We assessed lumbar spine bone mass (dual energy X-ray absorptiometry), distal tibia and radius bone microarchitecture and strength (high-resolution peripheral quantitative computed tomography), muscle function (jumping mechanography, dynamometry) and clinical outcomes (joint assessment, function, health-related quality of life) at baseline, 6- and 12-months. Adherence was assessed using weekly activity logs. Thirteen children completed the 6-month intervention. Participants reported 9 adverse events and post-exercise pain was rare (0.4%). Fatigue improved, but there were no other sustained improvements in muscle, bone or clinical outcomes. Adherence to the exercise program was low (47%) and decreased over time. Children with JIA safely participated in a home-based exercise program designed to enhance muscle and bone strength. Fatigue improved, which may in turn facilitate physical activity participation. Prescribed exercise posed adherence challenges and efforts are needed to address facilitators and barriers to participation in and adherence to exercise programs among children with JIA. Data of the children with JIA are from the LEAP study (Canadian Institutes of Health Research (CIHR; GRANT# 107535 ). http://www.leapjia.com/.


Light emission in nonlocal plasmonic metamaterials.
We present analytical and computational studies of light emission in nonlocal metamaterials formed by arrays of aligned plasmonic nanowires. We demonstrate that the emission lifetime in these composites is a complex function of geometrical and material parameters of the system that cannot be reduced to the "trivial" hyperbolic or elliptical dispersion topology of a homogenised metamaterial. In particular, our studies suggest that the Purcell factor can often be maximized when the composite operates in the elliptic regime, with strong radiation coupling to an additional TM-polarized mode supported by the nonlocal composite, in contrast to the accepted "hyperbolicity related" enhancement.