Atomic force microscopic investigation of respiratory syncytial virus infection in HEp-2 cells.
Respiratory syncytial virus (RSV) primarily causes bronchiolitis and pneumonia in infants. In spite of intense research, no safe and effective vaccine has been developed yet. For understanding its pathogenesis and development of anti-RSV drugs/therapeutics, it is indispensable to study the RSV-host interaction. Although, there are limited studies using electron microscopy to elucidate the infection process of RSV, to our knowledge, no study has reported the morphological impact of RSV infection using atomic force microscopy. We report the cytoplasmic and nuclear changes in human epidermoid cell line type 2 using atomic force microscopy. Human epidermoid cell line type 2 cells, grown on cover slips, were infected with RSV and fixed after various time periods, processed and observed for morphological changes using atomic force microscopy. RSV infected cells showed loss of membrane integrity, with degeneration in the cellular content and cytoskeleton. Nuclear membrane was disintegrated and nuclear volume was decreased. The chromatin of the RSV infected cells was condensed, progressing towards degeneration via pyknosis and apoptosis. Membrane protrusions of ~150-200 nm diameter were observed on RSV infected cells after 6 h, suggestive of prospective RSV budding sites. To our knowledge, this is the first study of RSV infection process using atomic force microscopy. Such morphological studies could help explore viral infection process aiding the development of anti-RSV therapies.


ICAM-1 and beta2 integrin deficiency impairs fat oxidation and insulin metabolism during fasting.
Intercellular adhesion molecule 1 (ICAM-1) and beta2 integrins play critical roles in immune responses. ICAM-1 may also participate in regulation of energy balance because ICAM-1-deficient mice become obese on a high-fat diet. We show that mice deficient in these adhesion receptors are unable to respond to fasting by up-regulation of fatty acid oxidation. Normal mice, when fasted, exhibit reduced circulating neutrophil counts and increased ICAM-1 expression and neutrophil recruitment in liver. Mice lacking ICAM-1 or beta2 integrins fail to show these responses--instead they become hypoglycemic with steatotic livers. Fasting ICAM-1-deficient mice reduce insulin more slowly than wild-type mice. This produces fasting hyperinsulinemia that prevents activation of adenosine mono-phosphate (AMP)-activated protein kinase in muscles and liver, which results in decreased import of long chain fatty acids into mitochondria. Thus, we show a new role for immune cells and their adhesion receptors in regulating metabolic response to fasting.


[Neuropeptides, Cytokines and Thymus Peptides as Effectors of Interactions Between Thymus and Neuroendocrine System].
The review presents data on mutual influence of nervous system and thymus, realized through the neuroendocrine-immune interactions. The pres- ence of adrenergic and peptidergic nerves in thymus creates conditions for implementation of the effect of neuropeptides secreted by them. These neuropeptides induce activation of thymus cells receptors and influence on the main processes in thymus, including T-lymphocyte maturation, cytokine and hormones production. In turn, thymuspeptides and/or cytokines, controlled by them, enter the brain and exert influence on neuro- nalfunction, which creates the basis for changes of behavior and homeostasis maintenance in response to infection. Ageing and some infectious, autoimmune, neurodegenerative and cancer diseases are accompanied by distortion of interactions between thymus and central nervous system. Mechanisms of signaling pathways, which determine these interactions, are not revealed yet, and their understanding will promote the development of effective therapeutic strategies.


Sickle cell disease and the general surgeon.
Sickle cell disease (SCD) is a common genetic disorder which represents a major medical problem in certain parts of the world. It is characterized by chronic haemolytic anaemia and vaso-occlusive crises, which can lead to widespread vascular occlusion by sickled red blood cells leading to multiple organ infarctions. In this respect, SCD can be considered as a multisystem disease presenting, to various surgical disciplines, a wide variety of surgical ailments. As surgical management of these patients is associated with high morbidity and mortality, close collaboration between medical and surgical services is essential. Greater population mobility makes recognition of the surgical implications of this disease of paramount importance in surgical practice. This article reviews surgical problems with which SCD patients may present to general surgeons and discusses their management.


Efficiency of ferredoxins and flavodoxins as mediators in systems for hydrogen evolution.
1. The efficiencies of ferredoxins and flavodoxins from a range of sources as mediators in systems for hydrogen evolution were assessed. 2. In supporting electron transfer from dithionite to hydrogenase of the bacterium Clostridium pasteurianum, highest activity was shown by the ferredoxin from the cyanobacterium Chlorogloeopsis fritschii and flavodoxin from the bacterium Megasphaera elsdenii. The latter was some twenty times as active as comparable concentrations of Methyl Viologen. Ferredoxins from the cyanobacterium Anacystis nidulans and the red alga Porphyra umbilicalis also showed high activity. 3. In mediating electron transfer from chloroplast membranes to Clostridium pasteurianum hydrogenase the flavodoxin from Anacystis nidulans proved the most active with Nostoc strain MAC flavodoxin and Porphyra umbilicalis ferredoxin also being appreciably more active than other cyanobacterial and higher plant ferredoxins. 4. In both hydrogenase systems the ferredoxin and flavodoxin from the red alga Chondrus crispus and the ferredoxin from another red alga Gigartina stellata showed very low activity. 5. There appeared to be no apparent correlation of efficiency in supporting hydrogenase activity with midpoint redox potential (Em) of the mediators, though some correlation of Em with the efficiency of the mediators in supporting NADP+ photoreduction by chloroplasts, or pyruvate oxidation by a Clostridium pasteurianum system, was evident. 6. Activity of the mediators in the hydrogenase systems therefore primarily reflects differences in tertiary structure conferring differing affinities for the other components of the systems.


Progression of Enamel Demineralisation Around Fissure Sealants: Optical Coherence Tomography Study.
To evaluate the progression of enamel demineralisation around fissure sealants using cross-polarisation optical coherence tomography (CP-OCT). Three fissure sealants; Fuji Triage (FJ, GC), Beautisealant (BT, Shofu) and Helioseal resin sealant (HL, Ivoclar Vivadent) were placed in cavities (3 × 0.5 × 1 mm) prepared in bovine enamel blocks (n = 5). After 7-day artificial saliva incubation, specimens were subjected to demineralisation for 4 weeks (pH 4.5). CP-OCT scans (Santec) were acquired for each specimen after 1, 2 and 4 weeks of demineralisation and lesions were quantitatively measured to analyse lesion progression. Repeated-measures ANOVA demonstrated that lesion size was statistically significantly affected by demineralisation period, fissure sealant type, and their interaction (P < 0.001). At 4 weeks, the highest mean value of lesion size was observed in HL group while FJ group showed the lowest. The rate of lesion progression was slower in FJ and statistically significantly different from BT (P < 0.05), which in turn was statistically significantly different than HL (P < 0.001). Fissure sealants that actively release ions are capable of improving the acid resistance of adjacent enamel. Beautisealant showed increased demineralisation inhibition compared to conventional resin sealant, but less than that provided by glass-ionomer sealant.


Ion channels as key partners of cytoskeleton in cancer disease.
Several processes occur during tumor development including changes in cell morphology, a reorganization of the expression and distribution of the cytoskeleton proteins as well as ion channels. If cytoskeleton proteins and ion channels have been widely investigated in understanding cancer mechanisms, the interaction between these two elements and the identification of the associated signaling pathways are only beginning to emerge. In this review, we summarize the work published over the past 15 years relating to the roles played by ion channels in these mechanisms of reorganization of the cellular morphology, essential to metastatic dissemination, both through the physical interactions with elements of the cytoskeleton and by intracellular signaling pathways involved.


A Placebo-Controlled Trial of Percutaneous Coronary Intervention for Stable Angina.
Percutaneous coronary intervention (PCI) is frequently performed to reduce the symptoms of stable angina. Whether PCI relieves angina more than a placebo procedure in patients who are not receiving antianginal medication remains unknown. We conducted a double-blind, randomized, placebo-controlled trial of PCI in patients with stable angina. Patients stopped all antianginal medications and underwent a 2-week symptom assessment phase before randomization. Patients were then randomly assigned in a 1:1 ratio to undergo PCI or a placebo procedure and were followed for 12 weeks. The primary end point was the angina symptom score, which was calculated daily on the basis of the number of angina episodes that occurred on a given day, the number of antianginal medications prescribed on that day, and clinical events, including the occurrence of unblinding owing to unacceptable angina or acute coronary syndrome or death. Scores range from 0 to 79, with higher scores indicating worse health status with respect to angina. A total of 301 patients underwent randomization: 151 to the PCI group and 150 to the placebo group. The mean (±SD) age was 64±9 years, and 79% were men. Ischemia was present in one cardiac territory in 242 patients (80%), in two territories in 52 patients (17%), and in three territories in 7 patients (2%). In the target vessels, the median fractional flow reserve was 0.63 (interquartile range, 0.49 to 0.75), and the median instantaneous wave-free ratio was 0.78 (interquartile range, 0.55 to 0.87). At the 12-week follow-up, the mean angina symptom score was 2.9 in the PCI group and 5.6 in the placebo group (odds ratio, 2.21; 95% confidence interval, 1.41 to 3.47; P<0.001). One patient in the placebo group had unacceptable angina leading to unblinding. Acute coronary syndromes occurred in 4 patients in the PCI group and in 6 patients in the placebo group. Among patients with stable angina who were receiving little or no antianginal medication and had objective evidence of ischemia, PCI resulted in a lower angina symptom score than a placebo procedure, indicating a better health status with respect to angina. (Funded by the National Institute for Health and Care Research Imperial Biomedical Research Centre and others; ORBITA-2 ClinicalTrials.gov number, NCT03742050.).


Adolescents' perceived benefits and barriers related to diabetes self-management--Part 1.
Knowledge is limited about adolescents' decision making regarding their assumption of diabetes management from parents. This study's purpose was to identify adolescents' perceived benefits and barriers about this process. Sixteen adolescents with type 1 diabetes were interviewed. Questions were adapted from the Janis and Mann (1977) decisional balance sheet. Interviews were audiotaped and transcribed verbatim. Descriptive expressions were identified and categories inductively generated. Adolescents' perceived benefits for themselves were having knowledge of or confidence in self-management abilities, more freedoms, and approval of others. They perceived benefits for parents as relief from responsibility, stress, and worry. Burden of responsibility was perceived as a barrier to adolescents for self-management. Adolescents perceived barriers for parents as worry/guilt and loss of control. Interrater agreement was initially 74%. Categories with disagreements were recategorized by interraters and then there was 100% agreement. Understanding adolescents' perceptions would be beneficial to their health care professionals.


Objective Structured Clinical Examination technical skill stations correlate more closely with postgraduate year level than do clinical skill stations.
Validity of an examination format is supported by its ability to distinguish levels of training among examinees. The Objective Structured Clinical Examination (OSCE) is a developing format generally composed of various types of task-oriented stations used to evaluate clinical skills of students and residents. The ideal composition of OSCE stations to maximize validity has not been determined. We examined the relative correlation between selected types of stations and level of resident postgraduate year (PGY). A 12-station OSCE was administered to surgical residents of all PGY levels at a university program. Individual station scores were correlated with PGY level. The overall correlation of the total examination score with PGY level was good (R = 0.681). Technical skill stations exhibited a significantly greater correlation with PGY level (0.679 vs 0.203) as compared with clinical skill stations (P < 0.05). These data suggest that technical skill evaluation is more sensitive in distinguishing level of training of surgical residents than is clinical skill evaluation.


Amino acid management of Parkinson's disease: a case study.
An extensive list of side effects and problems are associated with the administration of l-dopa (l-3, 4-dihydroxyphenylalanine) during treatment of Parkinson's disease. These problems can preclude achieving an optimal response with l-dopa treatment. To present a case study outlining a novel approach for the treatment of Parkinson's disease that allows for management of problems associated with l-dopa administration and discusses the scientific basis for this treatment. The case study was selected from a database containing 254 Parkinson's patients treated in developing and refining this novel approach to its current state. The spectrum of patients comprising this database range from newly diagnosed, with no previous treatment, to those who were diagnosed more than 20 years before and had virtually exhausted all medical treatment options. Parkinson's disease is associated with depletion of tyrosine hydroxylase, dopamine, serotonin, and norepinephrine. Exacerbating this is the fact that administration of l-dopa may deplete l-tyrosine, l-tryptophan, 5-hydroxytryptophan (5-HTP), serotonin, and sulfur amino acids. The properly balanced administration of l-dopa in conjunction with 5-HTP, l-tyrosine, l-cysteine, and cofactors under the guidance of organic cation transporter functional status determination (herein referred to as "OCT assay interpretation") of urinary serotonin and dopamine, is at the heart of this novel treatment protocol. When 5-HTP and l-dopa are administered in proper balance along with l-tyrosine, l-cysteine, and cofactors under the guidance of OCT assay interpretation, the long list of problems that can interfere with optimum administration of l-dopa becomes controllable and manageable or does not occur at all. Patient treatment then becomes more effective by allowing the implementation of the optimal dosing levels of l-dopa needed for the relief of symptoms without the dosing value barriers imposed by side effects and adverse reactions seen in the past.


Machine-Learning Classification of a Number of Contaminant Sources in an Urban Water Network.
In the case of a contamination event in water distribution networks, several studies have considered different methods to determine contamination scenario information. It would be greatly beneficial to know the exact number of contaminant injection locations since some methods can only be applied in the case of a single injection location and others have greater efficiency. In this work, the Neural Network and Random Forest classifying algorithms are used to predict the number of contaminant injection locations. The prediction model is trained with data obtained from simulated contamination event scenarios with random injection starting time, duration, concentration value, and the number of injection locations which varies from 1 to 4. Classification is made to determine if single or multiple injection locations occurred, and to predict the exact number of injection locations. Data was obtained for two different benchmark networks, medium-sized network Net3 and large-sized Richmond network. Additionally, an investigation of sensor layouts, demand uncertainty, and fuzzy sensors on model accuracy is conducted. The proposed approach shows excellent accuracy in predicting if single or multiple contaminant injections in a water supply network occurred and good accuracy for the exact number of injection locations.


[Primary prevention of hypertension in childhood (the population approach)].
A substantial prevalence of clinical hypertension with a large portion of the adult population requiring daily antihypertensive medication makes this disease an important public health problem. The known and unknown side effects of life-long pharmacologic therapy directs searches for methods of prevention. Because the origins of high blood pressure are found in children, at a point in life where health habits are established, preventive programs directed to youth seem particularly relevant. Increased sodium intake is the most widespread nutritional factor related to the development of high blood pressure. The prevalence of hypertension and the ubiquitous nature of sodium in many processed foods, well in excess of that needed for life seem to indicate the need for a broad public health approach. A test of this approach in school children is described below. The overall goals are to better understand the nature of health education and nutrition education in this age and, if successful, evaluate the effect of moderate sodium reduction on blood pressure.


Mathematical Modeling of Optical Radiation Emission as a Function of Welding Power during Gas Shielded Metal Arc Welding.
Arc welding is accompanied by intense optical radiation emission that can be detrimental not only for the welder himself but also for people working nearby or for passersby. Technological progress advances continuously in the field of joining, so an up-to-date radiation database is necessary. Additionally, many literature irradiance data have been measured for a few welding currents or for parts of the optical spectral region only. Within this paper, a comprehensive study of contemporary metal active gas, metal inert gas, and cold metal transfer welding is presented covering optical radiation emission from 200 up to 2,700 nm by means of (spectro-) radiometric measurements. The investigated welding currents range from 70 to 350 A, reflecting values usually applied in industry. Based upon these new irradiance data, three mathematical models were derived in order to describe optical radiation emission as a function of welding power. The linear, exponential, and sigmoidal emission models depend on the process variant (standard or pulsed) as well as on the welding material (mild and stainless steel, aluminum). In conjunction with the corresponding exposure limit values for incoherent optical radiation maximum permissible exposure durations were calculated as a function of welding power. Typical times are shorter than 1 s for the ultraviolet spectral region and range from 1 to 10 s for visible radiation. For the infrared regime, exposure durations are of the order of minutes to hours. Finally, a validation of the metal active gas emission models was carried out with manual arc welding.


Evaluation of microwave steam bags for the decontamination of filtering facepiece respirators.
Reusing filtering facepiece respirators (FFRs) has been suggested as a strategy to conserve available supplies for home and healthcare environments during an influenza pandemic. For reuse to be possible, used FFRs must be decontaminated before redonning to reduce the risk of virus transmission; however, there are no approved methods for FFR decontamination. An effective method must reduce the microbial threat, maintain the function of the FFR, and present no residual chemical hazard. The method should be readily available, inexpensive and easily implemented by healthcare workers and the general public. Many of the general decontamination protocols used in healthcare and home settings are unable to address all of the desired qualities of an efficient FFR decontamination protocol. The goal of this study is to evaluate the use of two commercially available steam bags, marketed to the public for disinfecting infant feeding equipment, for FFR decontamination. The FFRs were decontaminated with microwave generated steam following the manufacturers' instructions then evaluated for water absorption and filtration efficiency for up to three steam exposures. Water absorption of the FFR was found to be model specific as FFRs constructed with hydrophilic materials absorbed more water. The steam had little effect on FFR performance as filtration efficiency of the treated FFRs remained above 95%. The decontamination efficacy of the steam bag was assessed using bacteriophage MS2 as a surrogate for a pathogenic virus. The tested steam bags were found to be 99.9% effective for inactivating MS2 on FFRs; however, more research is required to determine the effectiveness against respiratory pathogens.


Neurophysiological and behavioral responses of blacklegged ticks to host odors.
The blacklegged tick, Ixodes scapularis (Ixodida, Ixodidae), is one of the major disease vectors in the United States, and due to multiple human impact factors, such as decreasing forest size for land development and climate change, it has expanded its range and established across the United States. Throughout the life cycle, ticks locate hosts for their blood-meal, and although the ecologies of this tick and their hosts have been studied in depth, the sensory physiology behind host location largely remains unexplored. Here, we report establishing a robust paradigm to isolate and identify odors from the natural milieu for I. scapularis. We performed single sensillum recordings (SSR) from the olfactory sensilla on the tick tarsi, and used the SSR system as a biological detector to isolate natural compounds that elicited biological activity. The SSR setup was further tested in tandem with gas chromatography (GC) wherein the ticks' olfactory sensillum activity served as a biological detector. The GC-SSR recordings from the wall pore sensilla in the Haller's organ, and further identification of the biologically active deer gland constituents by GC-mass spectrometry (GC-MS) revealed methyl substituted phenols as strong chemostimuli, as compared to ethyl or propyl substitutions. The strongest electrophysiological activity was elicited by m- cresol followed by p- cresol. Ethyl- and propylphenols with any of the three substitutions (ortho, meta or para), did not induce any neurophysiological activity. Finally, a behavioral analysis in a dual-choice olfactometer of all these phenols at three different doses revealed no significant behavioral response, except for p- cresol at -3 dilution. Overall, this study contributes to our understanding of I. scapularis tick's neurophysiology and provides a robust platform to isolate and identify natural attractants and repellents.


Support Immersion Endoscopy in Post-Extraction Alveolar Bone Chambers: A New Window for Microscopic Bone Imaging In Vivo.
Using an endoscopic approach, small intraoral bone chambers, which are routinely obtained during tooth extraction and implantation, provide visual in vivo access to internal bone structures. The aim of the present paper is to present a new method to quantify bone microstructure and vascularisation in vivo. Ten extraction sockets and 6 implant sites in 14 patients (6 men / 8 women) were examined by support immersion endoscopy (SIE). After tooth extraction or implant site preparation, microscopic bone analysis (MBA) was performed using short distance SIE video sequences of representative bone areas for off-line analysis with ImageJ. Quantitative assessment of the microstructure and vascularisation of the bone in dental extraction and implant sites in vivo was performed using ImageJ. MBA revealed bone morphology details such as unmineralised and mineralised areas, vascular canals and the presence of bleeding through vascular canals. Morphometric examination revealed that there was more unmineralised bone and less vascular canal area in the implant sites than in the extraction sockets.


Treatment of severe beta-thalassemia (patients) with myleran.
We previously reported that myleran, a cell cycle nonspecific drug, can stimulate gamma-globin gene expression in anemic adult rhesus monkeys. This finding prompted us to treat two patients with severe beta-thalassemia with myleran. Both patients received an initial course of therapy, constantly of myleran at a dosage of 0.2 mg/kg/d for 9 days followed by 0.15 mg/kg/d for the next 11 days. One patient received an additional 20-day course of myleran at a dosage of 0.2 mg/kg/d beginning 44 days after completion of the first course. No severe ill effects related to the drug were observed during or after drug administration. After 20 days of myleran treatment, levels of HbF and reticulocytes increased in both patients and level of F cells increased in patient 1. In patient 1, Hb concentration rose from 42g/L (9 days after transfusion) before treatment to a maximum of 65g/L afterward; in patient 2, it rose from 74g/L to a maximum of 106g/L. A value of 15g/L above baseline lasted for about 5 months in both patients. Hypomethylation of bone marrow DNA near the gamma-globin gene was demonstrated in patient 1.


Inducer expulsion and the occurrence of an HPr(Ser-P)-activated sugar-phosphate phosphatase in Enterococcus faecalis and Streptococcus pyogenes.
Inducer expulsion, a phenomenon in which rapidly metabolizable sugars cause cytoplasmic dephosphorylation and efflux of pre-accumulated sugar-phosphates (sugar-P), has been documented for Streptococcus pyogenes, Streptococcus bovis, and Lactococcus lactis, but not for other Gram-positive bacteria. Using intact cells and membrane vesicles, we show that Enterococcus faecalis exhibits both inducer exclusion and inducer expulsion, and that the latter phenomenon is dependent on the metabolite-activated ATP-dependent HPr(Ser) kinase that phosphorylates Ser-46 in HPr of the phosphotransferase system. A small, heat-stable, membrane-associated, HPr(Ser-P)-activated sugar-P phosphatase (Pase II), previously identified only in Lc. lactis, is shown to be present in extracts of Enterococcus faecalis and Streptococcus pyogenes but not in those of Staphylococcus aureus, Streptococcus mutans, Streptococcus salivarius, or Bacillis subtilis, organisms that do not exhibit the inducer expulsion phenomenon. Further, Lactobacillus brevis, an organism that exhibits inducer expulsion by a different mechanism, also apparently lacks Pase II. The results reveal that Pase II is present in those organisms that exhibit the coupled sugar-P hydrolysis/expulsion mechanism but not those that lack this mechanism. They provide correlative evidence that Pase II initiates inducer expulsion in species of enterococci, streptococci and lactococci.


[Studies on the constituents of cucurbitaceous plants].
I have an opportunity to have co-operative studies with Chinese group of Kunming Institute of Botany. Since then, I have investigated the chemical constituents of a number of Chinese plants of Araliaceae, Umbelliferae, Labiatae, Cucurbitaceae and other families. This review describes the structural elucidation of the cucurbitane, oleanane and dammarane glycosides, and their biological activities under the joint studies on cucurbitaceous plants, Bolbostemma, Hemsleya, Siraitia and Neoalsomitra species. New oleanane glycosides having novel cyclic structure were isolated from Bolbostemma paniculatum. The potent solubilizing effect of these compounds was investigated. A number of cucurbitane glycosides were isolated from Hemsleya carnosiflora, H. panacis-scandens, Siraitia grosvenorii and S. siamensis. Some of these glycosides taste sweet, bitter or tasteless. The structure-taste relationships of the glycosides of a 3-alpha-hydroxy-cucurbit-5-ene-type triterpene have been discussed. Anti-tumor-promotion effects as well as the ecdysteroid agonist and antagonist activities of these cucurbitane glycosides were investigated. New dammarane glycosides were isolated from Neoalsomitra integrifoliola.


A school fluoride varnish program for Aboriginal children in rural New South Wales, Australia.
Australian Aboriginal children are less likely to access preventive oral health services such as fluoride varnish. The regular application of fluoride varnish can prevent dental caries. This study aims to determine if a school-based fluoride varnish program can provide Aboriginal children with at least three fluoride varnish applications over 12 months and whether the routine application of fluoride varnish for Aboriginal children in schools is a feasible approach for oral health promotion in Aboriginal communities. A school fluoride varnish program was co-designed with Aboriginal communities in Central Northern New South Wales, Australia and implemented in three schools where majority of enrolled children are Aboriginal. Four "fluoride varnish days," 3 months apart, were held at each school over the 12-month study period. On each "fluoride varnish day" an oral health therapist applied fluoride varnish to all children included in the study. This study took place between January and December 2017. About 153 children were eligible to participate and 131 (86%) were consented into the program by a parent or guardian. A total of 104 children were enrolled for the entire 12-month study period and included in the analysis. Majority of children (65.4%) received at least three fluoride varnish applications, with an average of 70% of students receiving an application of fluoride varnish on each "fluoride varnish day." School-based fluoride varnish programs, co-designed with local Aboriginal communities, may be a feasible approach to oral health promotion aimed at improving the oral health of Aboriginal children. This study aimed to improve access to fluoride varnish for Aboriginal children in Central Northern NSW. Four "fluoride varnish days," each three months apart, were held in three schools over 12 months. Majority of children (65.4%) received at least three fluoride varnish applications during the fluoride varnish program.


Predicting the Naturalistic Course of Major Depressive Disorder Using Clinical and Multimodal Neuroimaging Information: A Multivariate Pattern Recognition Study.
A chronic course of major depressive disorder (MDD) is associated with profound alterations in brain volumes and emotional and cognitive processing. However, no neurobiological markers have been identified that prospectively predict MDD course trajectories. This study evaluated the prognostic value of different neuroimaging modalities, clinical characteristics, and their combination to classify MDD course trajectories. One hundred eighteen MDD patients underwent structural and functional magnetic resonance imaging (MRI) (emotional facial expressions and executive functioning) and were clinically followed-up at 2 years. Three MDD trajectories (chronic n = 23, gradual improving n = 36, and fast remission n = 59) were identified based on Life Chart Interview measuring the presence of symptoms each month. Gaussian process classifiers were employed to evaluate prognostic value of neuroimaging data and clinical characteristics (including baseline severity, duration, and comorbidity). Chronic patients could be discriminated from patients with more favorable trajectories from neural responses to various emotional faces (up to 73% accuracy) but not from structural MRI and functional MRI related to executive functioning. Chronic patients could also be discriminated from remitted patients based on clinical characteristics (accuracy 69%) but not when age differences between the groups were taken into account. Combining different task contrasts or data sources increased prediction accuracies in some but not all cases. Our findings provide evidence that the prediction of naturalistic course of depression over 2 years is improved by considering neuroimaging data especially derived from neural responses to emotional facial expressions. Neural responses to emotional salient faces more accurately predicted outcome than clinical data.


Appropriateness and preferential use of different seasonal influenza vaccines: A pilot study on the opinion of vaccinating physicians in Italy.
In Italy, several types of influenza vaccine are on the market. Available evidence suggests that no single vaccine type is universally appropriate; rather, different types may be more appropriate for different population strata. However, while the concept of appropriateness/preferential use of single vaccines is usually adopted at the central level, little is known about the attitudes of physicians on the matter. A pilot survey of Italian physicians (N = 372) revealed that most (about 90%) were aware that the available vaccines were different, and that particular vaccines were more appropriate for specific groups. The availability of explicit guidelines on which vaccine to administer to a given population group was deemed desirable by 93.2% of respondents. The results were consistent with the 2018 Italian and UK normative documents, which indicate adjuvanted vaccines as the most appropriate choice for the elderly, and quadrivalent formulations for the younger age-classes. Public health policy implications are discussed.


Calmodulinopathy: A Novel, Life-Threatening Clinical Entity Affecting the Young.
Sudden cardiac death (SCD) in the young may often be the first manifestation of a genetic arrythmogenic disease that had remained undiagnosed. Despite the significant discoveries of the genetic bases of inherited arrhythmia syndromes, there remains a measurable fraction of cases where in-depth clinical and genetic investigations fail to identify the underlying SCD etiology. A few years ago, 2 cases of infants with recurrent cardiac arrest episodes, due to what appeared to be as a severe form of long QT syndrome (LQTS), came to our attention. These prompted a number of clinical and genetic research investigations that allowed us to identify a novel, closely associated to LQTS but nevertheless distinct, clinical entity that is now known as calmodulinopathy. Calmodulinopathy is a life-threatening arrhythmia syndrome, affecting mostly young individuals, caused by mutations in any of the 3 genes encoding calmodulin (CaM). Calmodulin is a ubiquitously expressed Ca2+ signaling protein that, in the heart, modulates several ion channels and participates in a plethora of cellular processes. We will hereby provide an overview of CaM's structure and function under normal and disease states, highlighting the genetic etiology of calmodulinopathy and the related disease mechanisms. We will also discuss the phenotypic spectrum of patients with calmodulinopathy and present state-of-the art approaches with patient-derived induced pluripotent stem cells that have been thus far adopted in order to accurately model calmodulinopathy in vitro, decipher disease mechanisms and identify novel therapies.


The vascular complications of diabetes: a review of their management, pathogenesis, and prevention.
This review highlights the pathogenesis of both microvascular and macrovascular complications of diabetes and how these mechanisms influence both the management and preventative strategies of these complications. The cumulative data shown in this review suggest hyperglycemic and blood pressure control remain central to this intricate process. We reviewed the literature including retrospective, prospective trials as well as meta-analysis, and post hoc analysis of randomized trials on microvascular andmacrovascular complications. Further research is needed to explore the ideal intervention targets and preventative strategies needed to prevent macrovascular complications. Furthermore, as the data for trials looking at microvascular complications lengthen more long-term data will further elucidate the role that the duration of diabetes has on these complications. Additionally, trials looking to maximize hyperglycemic control with multiple agents in diabetes, such as metformin, SGL2isand GLP-1 receptor agonists are currently in process, which will have implications for rates of microvascular as well as macrovascular complications.


Germline and somatic DNA repair gene alterations in prostate cancer.
Emerging evidence has suggested that DNA repair gene alterations may be important in prostate cancer pathogenesis. In the current study, the authors sought to characterize alterations in DNA repair pathway genes in both primary and metastatic prostate tumors with attention to tissue distribution as well as specific genomic alterations. The authors studied the distribution and type of alterations in 24 genes that are considered important for DNA repair in 944 prostate cancers harvested from localized and metastatic tumors. Tumor DNA underwent hybrid capture for all coding exons of 287 or 395 cancer-related genes plus select introns from 19 or 31 genes frequently rearranged in cancer. Captured libraries were sequenced to a median exon coverage depth of >×500. Specific genomic alterations were characterized and the frequencies of mutations by tissue site (prostate vs metastases) were compared using logistic regression. A total of 152 patients from the cohort of 944 men (16%) harbored a germline or somatic mutation in ≥1 DNA repair genes. The most frequently mutated genes were BRCA2 (11.4%) and ATM (5.8%), followed by MSH6 (2.5%) and MSH2 (2.1%). Mutations were identified in approximately 20.1% of primary prostate tumors compared with 18.8% of bone metastases. When stratified by tissue site, the highest rates of DNA repair mutations were found in solid organ metastases, including brain and visceral metastases, compared with prostate. DNA repair gene mutations are more common in metastatic than localized prostate tumors. Visceral and other solid organ metastases appear enriched for these mutations compared with localized tumors or bone and lymph node metastases.


Stakeholder perspectives on the demand and supply factors driving substandard and falsified blood pressure lowering medications in Nigeria: a qualitative study.
Although substandard and falsified (SF) blood pressure (BP) lowering medications are a global problem, qualitative research exploring factors driving this in Nigeria has not been reported. This study provides information on factors driving demand for and supply of low-quality BP lowering medications in Nigeria and potential strategies to address these factors. This was a cross-sectional qualitative study. Between August 2020 and September 2020, we conducted 11 in-depth interviews and 7 focus group discussions with administrators of health facilities, major manufacturers and distributors of BP lowering medications, pharmacists, drug regulators, patients and primary care physicians purposively sampled from the Federal Capital Territory, Nigeria. Data were analysed using directed content analysis, with the aid of Dedoose. We found that demand for SF BP lowering medications in Nigeria was driven by high out-of-pocket expenditure and stockouts of quality-assured BP lowering medications. Supply of low-quality BP lowering medications was driven by limited in-country manufacturing capacity, non-adherence to good manufacturing and distribution practices, under-resourced drug regulatory systems, ineffective healthcare facility operations, poor distribution practices, limited number of trained pharmacists and the COVID-19 pandemic which led to stockouts. Central medicine store procurement procedures, active pharmaceutical ingredient quality check and availability of trained pharmacists were existing strategies perceived to lower the risk of supply and demand of SF BP lowering medications. Our findings suggest that demand for and supply of SF BP lowering medications in Nigeria are driven by multi-level, interrelated factors. Multi-pronged strategies need to target stakeholders and systems involved in drug production, distribution, prescription, consumption, regulation and pricing.


Highly Specific Sewage-Derived Bacteroides Quantitative PCR Assays Target Sewage-Polluted Waters.
The identification of sewage contamination in water has primarily relied on the detection of human-associated Bacteroides using markers within the V2 region of the 16S rRNA gene. Despite the establishment of multiple assays that target the HF183 cluster (i.e., Bacteroides dorei) and other Bacteroides organisms (e.g., Bacteroides thetaiotaomicron), the potential for more human-associated markers in this genus has not been explored in depth. We examined the Bacteroides population structure in sewage and animal hosts across the V4V5 and V6 hypervariable regions. Using near-full-length cloned sequences, we identified the sequences in the V4V5 and V6 hypervariable regions that are linked to the HF183 marker in the V2 region and found these sequences were present in multiple animals. In addition, the V4V5 and V6 regions contained human fecal marker sequences for organisms that were independent of the HF183 cluster. The most abundant Bacteroides in untreated sewage was not human associated but pipe derived. Two TaqMan quantitative PCR (qPCR) assays targeting the V4V5 and V6 regions of this organism were developed. Validation studies using fecal samples from seven animal hosts (n = 76) and uncontaminated water samples (n = 30) demonstrated the high specificity of the assays for sewage. Freshwater Bacteroides were also identified in uncontaminated water samples, demonstrating that measures of total Bacteroides do not reflect fecal pollution. A comparison of two previously described human Bacteroides assays (HB and HF183/BacR287) in municipal wastewater influent and sewage-contaminated urban water samples revealed identical results, illustrating the assays target the same organism. The detection of sewage-derived Bacteroides provided an independent measure of sewage-impacted waters.IMPORTANCEBacteroides are major members of the gut microbiota, and host-specific organisms within this genus have been used extensively to gain information on pollution sources. This study provides a broad view of the population structure of Bacteroides within sewage to contextualize the well-studied HF183 marker for a human-associated Bacteroides The study also delineates host-specific sequence patterns across multiple hypervariable regions of the 16S rRNA gene to improve our ability to use sequence data to assess water quality. Here, we demonstrate that regions downstream of the HF183 marker are nonspecific but other potential human-associated markers are present. Furthermore, we show the most abundant Bacteroides in sewage is free living, rather than host associated, and specifically found in sewage. Quantitative PCR assays that target organisms specific to sewer pipes offer measures that are independent of the human microbiome for identifying sewage pollution in water.


Electrophoretic evaluation of sera from dogs with cancer.
A quantitative electrophoretic method was developed for evaluating sera from dogs with naturally occurring malignant tumors. Electrophoretic migration ratios (EMR) were devised to standardize characteristic protein components and to analyze alterations in mobility of these protein components, using a computerized densitometric technique. Sera values from dogs with lymphosarcomas, osteosarcomas, and mammary tumors were compared, using conventional methods of statistical analysis. Results indicate a statistical difference existed between mean values of EMR and alpha2-globulins of sera from dogs with tumors and those values from normal dogs. In addition, unique EMR and concentration of protein components were identified for dogs with lymphosarcomas, osteosarcomas, and mammary tumors.


Solvent/detergent-treated plasma has decreased antitrypsin activity and absent antiplasmin activity.
Solvent/detergent (S/D)-treated plasma is currently marketed by the American Red Cross as a virally inactivated alternative to fresh-frozen plasma (FFP). The serpin-type serine proteinase inhibitors have a flexible reactive site loop (RSL) that can convert from the active conformation to the inactive latent or polymerized conformations when exposed to heat and/or detergents. We have compared the conformational stability and inhibitory activity of 3 plasma serpins-antithrombin, antitrypsin, and antiplasmin-in S/D plasma and FFP. In S/D plasma, virtually 100% of the antiplasmin and approximately 50% of the antitrypsin are in either the latent or polymerized conformation and lack inhibitory activity, while in FFP only the active conformation is present. Interestingly, antithrombin is not affected by S/D treatment and remains fully active. These data demonstrate that S/D plasma is not simply a virally inactivated equivalent of FFP. The lack of antiplasmin activity and decreased antitrypsin activity in S/D plasma suggest that it may not be as effective as FFP for the treatment of bleeding in patients with systemic activation of proteolytic cascades, such as disseminated intravascular coagulation and sepsis, acquired fibrinolytic states, and large-volume transfusion. Although there has been extensive use of S/D plasma in several European countries with no reports of adverse effects, clinical studies directly comparing the efficacy of these 2 plasma products are needed to directly evaluate the relative therapeutic efficacy of FFP and S/D plasma for the treatment of these diseases.


The usual suspects, dopamine and alpha-synuclein, conspire to cause neurodegeneration.
Parkinson's disease (PD) is primarily a movement disorder driven by the loss of dopamine-producing neurons in the substantia nigra (SN). Early identification of the oxidative properties of dopamine implicated it as a potential source of oxidative stress in PD, yet few studies have investigated dopamine neurotoxicity in vivo. The discovery of PD-causing mutations in α-synuclein and the presence of aggregated α-synuclein in the hallmark Lewy body pathology of PD revealed another important player. Despite extensive efforts, the precise role of α-synuclein aggregation in neurodegeneration remains unclear. We recently manipulated both dopamine levels and α-synuclein expression in aged mice and found that only the combination of these 2 factors caused progressive neurodegeneration of the SN and an associated motor deficit. Dopamine modified α-synuclein aggregation in the SN, resulting in greater abundance of α-synuclein oligomers and unique dopamine-induced oligomeric conformations. Furthermore, disruption of the dopamine-α-synuclein interaction rescued dopaminergic neurons from degeneration in transgenic Caenorhabditis elegans models. In this Perspective, we discuss these findings in the context of known α-synuclein and dopamine biology, review the evidence for α-synuclein oligomer toxicity and potential mechanisms, and discuss therapeutic implications. © 2019 International Parkinson and Movement Disorder Society.


The Role and Significance of Wnt5a in Regulating Epithelial-Mesenchymal Transition in Endometrioid Adenocarcinoma.
As a non-classical ligand of Wnt, the abnormal regulation of Wnt5a contributes to the progression of malignant tumors; however, its effects differ depending on tumor type. Here, we evaluated the expression and significance of Wnt5a in endometrioid adenocarcinoma and its relationship with epithelial-mesenchymal transition (EMT)-related proteins. Immunohistochemical streptavidin-peroxidase method and reverse transcription polymerase chain reaction (RT-PCR) method were used to analyze the expression and correlation of Wnt5a, and EMT-related protein β-catenin, E-cadherin and enhancer of zeste homolog 2 (EZH2) in endometrial cancer tissues and cell samples of each group. The expression of Wnt5a and E-cadherin decreased in the following order, normal endometrium > atypical hyperplasia endometrium > endometrioid adenocarcinoma. In contrast, the expression of β-catenin and EZH2 increased gradually. Moreover, Wnt5a expression was associated with the degree of tissue differentiation, International Federation of Gynecology and Obstetrics (FIGO) stage, and lymph node metastasis (all P<0.05). Wnt5a expression was also negatively correlated with β-catenin and EZH2 expression and positively correlated with E-cadherin expression. RT-PCR results further indicated that E-cadherin mRNA expression was upregulated in a Wnt5a-overexpressing Ishikawa cell line compared to cells transfected with an empty vector or negative control cells (P<0.01). Furthermore, the expression of EZH2 and β-catenin mRNA was downregulated in overexpressing cells compared to empty vector and negative control cells (P<0.01). Wnt5a may elicit a suppressive effect on endometrioid adenocarcinoma by inhibiting EMT. This study provides a theoretical basis for the pathological diagnosis and targeted therapy of endometrioid adenocarcinoma and extends our understanding of the Wnt5a signaling pathway.


The control of ribonucleic acid synthesis in bacteria. Fluctuations in messenger ribonucleic acid synthesis in cultures recovering from amino acid starvation.
Changes in the cell content and rate of synthesis of mRNA were studied in auxotrophs of Escherichia coli recovering from a period of amino acid deprivation. Parallel studies were carried out on bacterial strains inhibited with trimethoprim, when glycine and methionine were added to relieve an amino acid deficiency. In the latter case, protein synthesis was still severely inhibited through a lack of N-formylmethionyl-tRNA(fMet) for chain initiation, so that fewer ribosomes were attached to mRNA chains. (1) In RC(str) strains recovering from amino acid starvation, there was a transient oversynthesis of mRNA, but the amounts returned to normal after about a 15-min period of recovery. RC(rel) strains did not show this effect; any extra mRNA accumulated during the previous starvation period was rapidly lost, but no oversynthesis occurred during the resumption of growth. (2) In trimethoprim-inhibited cultures supplemented with glycine and methionine, mRNA was produced at the same rate, relative to stable RNA species, as during normal growth. The evidence implied that decreased rates of ribosome attachment had no effect on the functional or chemical lifetime of the mRNA fraction. This suggests that mRNA stability does not depend on the frequency of translation by ribosomes. (3) Changes in the mRNA contents of trimethoprim-inhibited RC(str) and RC(rel) cultures were noted soon after supplementation with glycine and methionine. These closely followed those observed in cultures recovering from simple amino acid withdrawal.


Mechanisms of nitric oxide synthase uncoupling in endotoxin-induced acute lung injury: role of asymmetric dimethylarginine.
Acute lung injury (ALI) is associated with severe alterations in lung structure and function and is characterized by hypoxemia, pulmonary edema, low lung compliance and widespread capillary leakage. Asymmetric dimethylarginine (ADMA), a known cardiovascular risk factor, has been linked to endothelial dysfunction and the pathogenesis of a number of cardiovascular diseases. However, the role of ADMA in the pathogenesis of ALI is less clear. ADMA is metabolized via hydrolytic degradation to l-citrulline and dimethylamine by the enzyme, dimethylarginine dimethylaminohydrolase (DDAH). Recent studies suggest that lipopolysaccharide (LPS) markedly increases the level of ADMA and decreases DDAH activity in endothelial cells. Thus, the purpose of this study was to determine if alterations in the ADMA/DDAH pathway contribute to the development of ALI initiated by LPS-exposure in mice. Our data demonstrate that LPS exposure significantly increases ADMA levels and this correlates with a decrease in DDAH activity but not protein levels of either DDAH I or DDAH II isoforms. Further, we found that the increase in ADMA levels cause an early decrease in nitric oxide (NO(x)) and a significant increase in both NO synthase (NOS)-derived superoxide and total nitrated lung proteins. Finally, we found that decreasing peroxynitrite levels with either uric acid or Manganese (III) tetrakis (1-methyl-4-pyridyl) porphyrin (MnTymPyp) significantly attenuated the lung leak associated with LPS-exposure in mice suggesting a key role for protein nitration in the progression of ALI. In conclusion, this is the first study that suggests a role of the ADMA/DDAH pathway during the development of ALI in mice and that ADMA may be a novel therapeutic biomarker to ascertain the risk for development of ALI.


Cognitive predictors of parent-rated inattention in very preterm children: The role of working memory and processing speed.
Inattention is one of the most common neurobehavioral problems following very preterm birth. Attention problems can persist into adulthood and are associated with negative socio-emotional and educational outcomes. This study aimed to determine whether the cognitive processes associated with inattention differ between term-born and very preterm children. Sixty-five children born very preterm (<33+0 weeks' gestation) aged 8-11 years were recruited alongside 48 term-born controls (?37 20 +0 weeks' gestation). Both groups included children with a wide spectrum of parent-rated inattention (above average attention to severe inattention) measured as a continuous dimension using the Strengths and Weaknesses of ADHD and Normal-Behavior (SWAN) scale. The children completed tests to assess basic cognitive processes and executive function. A hierarchical multiple regression analysis was implemented to assess which neurocognitive processes explained variance in parent-rated inattention and whether these differed between preterm and term-born children. In both groups, poorer verbal and visuospatial short-term memory and poorer visuospatial working memory independently explained variance in parent-rated inattention. Slower motor processing speed explained variance in inattention among very preterm children only. The cognitive mechanisms associated with parent-rated inattention were predominantly overlapping between groups, but relationships between motor processing speed and inattention were unique to very preterm children. These associations may reflect risk factors for inattention in term and very preterm children. Future research should assess the efficacy of these cognitive processes as potential targets for intervention.


Possible involvement of regulatory T cell abnormalities and variational usage of TCR repertoire in children with autoimmune neutropenia.
Autoimmune neutropenia (AIN) in childhood is characterized by chronic neutropenia and positivity for anti-neutrophil antibodies, resulting in the excessive destruction of neutrophils. In this study, we investigated the involvement of regulatory T cells (Tregs ) in the pathogenesis of AIN in childhood. Tregs have been classified into three subpopulations based on the expressions of CD45RA and forkhead box protein 3 (FoxP3): resting Tregs , activated Tregs and non-suppressive Tregs . The frequency of activated Tregs (CD4+ CD25+ FoxP3high CD45RA- T cells) as well as that of total Tregs (CD4+ CD25+ FoxP3+ T cells) in peripheral blood was significantly decreased in patients with AIN. Analysis of the T cell receptor (TCR)-Vβ repertoire of CD4+ T cells revealed skewed usages in patients with AIN compared with that observed in age-matched control subjects. Regarding T cell subsets, the use of four of 24 TCR-Vβ families in Tregs and one in conventional T cells were increased in patients with AIN. The number of patients with AIN who showed skewed usages of TCR-Vβ family in conventional and Tregs was significantly higher than that reported in control subjects. When the preference between Tregs and conventional T cells in each TCR-Vβ family was individually compared, different use was prominently observed in the TCR-Vβ 9 family in patients with AIN. These results suggest that the quantitative abnormalities of Tregs and the skew of the TCR-Vβ repertoire in CD4+ T cells, including Tregs and conventional T cells, may be related to autoantibody production through a human neutrophil antigen-reactive T cell clone.


Endoscopic Ultrasound-Guided Biliary Access, with Focus on Technique and Practical Tips.
In 1980, endoscopic ultrasound (EUS) was introduced as a diagnostic tool for evaluation of the pancreas. Since the introduction of curvilinear-array echoendoscopy, EUS has been used for a variety of gastrointestinal interventions, including fine needle aspiration, tumor ablation, and pancreatobiliary access. One of the main therapeutic roles of EUS is biliary drainage as an alternative to endoscopic retrograde biliary drainage (ERBD) or percutaneous transhepatic biliary drainage (PTBD). This article summarizes three different methods of EUS-guided biliary access, with focus on technique and practical tips.


Label-free integrative pharmacology on-target of opioid ligands at the opioid receptor family.
In vitro pharmacology of ligands is typically assessed using a variety of molecular assays based on predetermined molecular events in living cells. Many ligands including opioid ligands pose the ability to bind more than one receptor, and can also provide distinct operational bias to activate a specific receptor. Generating an integrative overview of the binding and functional selectivity of ligands for a receptor family is a critical but difficult step in drug discovery and development. Here we applied a newly developed label-free integrative pharmacology on-target (iPOT) approach to systematically survey the selectivity of a library of fifty-five opioid ligands against the opioid receptor family. All ligands were interrogated using dynamic mass redistribution (DMR) assays in both recombinant and native cell lines that express specific opioid receptor(s). The cells were modified with a set of probe molecules to manifest the binding and functional selectivity of ligands. DMR profiles were collected and translated to numerical coordinates that was subject to similarity analysis. A specific set of opioid ligands were then selected for quantitative pharmacology determination. Results showed that among fifty-five opioid ligands examined most ligands displayed agonist activity in at least one opioid receptor expressing cell line under different conditions. Further, many ligands exhibited pathway biased agonism. We demonstrate that the iPOT effectively sorts the ligands into distinct clusters based on their binding and functional selectivity at the opioid receptor family.


Effects of 4-nonylphenol on balance of steroid and thyroid hormones in sexually immature male yellowfin seabream (Acanthopagrus latus).
Nonylphenol (NP) is an endocrine disrupting chemical which has been shown to be able to modulate the endocrine system of various organisms by different mechanisms. The objective of this study was to investigate the potential effects of 4-NP on steroid and thyroid hormone levels in sexually immature male yellowfin seabream (Acanthopagrus latus), a protandrous hermaphrodite species. For this, the fish were injected with ascending doses (10, 50, 100, and 200 μg g(-1) body weight) of 4-nonylphenol (4-NP) or vehicle during 2 weeks. After 7 and 14 days the fish were anesthetized, blood sample were collected and plasma steroid and thyroid hormone concentrations were quantified by radioimmunoassay. The result showed that 4-NP induced a significant increase in 17β-etradiol levels at dose 10 μg g(-1) , while the levels of this hormone in the higher doses decreased compared with the control group. However, 4-NP treatment did not have any significant effect on plasma levels of testosterone. In addition, it was observed that 4-NP affect the level of thyroid hormones in fish. Plasma thyroxine levels increased in a dose-dependent manner after 7 and 14 days of the exposure. In contrast, a significant decrease in triiodothyronine levels was observed during the experiment period. Moreover, no significant change was detected for thyroid stimulating hormone levels in 4-NP-treated fish. These results indicated that 4-NP could lead to disturb the balance of steroid and thyroid hormones with potential consequences for sexually immature male yellowfin seabream.


Strategies for the identification of loci responsible for the pathogenesis of multiple sclerosis.
Multiple sclerosis (MS) is a chronic, debilitating disease, which manifests itself by de-myelination of the central nervous system (CNS). MS is predominantly found in Caucasians of European decent and is more prominent in females than males. MS is one of the most prevalent causes of disability of young adults in the world. The exact cause of MS is not known, however genetic susceptibility to MS is linked to the major histocompability complex (MHC). Self reactive CD4+ T cells, specific for CNS antigens, such as myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG) and proteolipid protein (PLP), are detectable in MS patients along with pathogenic autoantibodies specific to these CNS antigens produced by B cells. These observations suggest that MS is an autoimmune disease. Epidemiology of MS along with the analysis of sibling pairs and twins suggest that the multiple genetic factors and their interaction with environment contribute to disease susceptibility. Recent developments and advancements in genetic analysis may aid in accurate determination of genetic risk factors for the development of MS. We review these developments, advances in technology and discuss recent results in this article.


Hypersensitivity to Carboplatin in Children with Malignancy.
Purpose: Carboplatin-based regimens have proven efficacy in children with cancer. However, the development of hypersensitivity reactions (HSRs) may have a negative impact on treatment intensity and patients' outcome. The aim of this review is to summarize the incidence and the clinical features of HSRs occurring in children with cancer treated with carboplatin and their impact on treatment efficacy. Methods: Data were collected by searching for relevant studies on the incidence, clinical features and management of possible side effects about the use of carboplatin in children, published from March 1987 to October 2016 in the PubMed database. Results: Carboplatin HSRs present with mild/moderate to severe clinical patterns. The risk of HSR is related to the cumulative number of infusions. Moreover, a greater risk of developing an HSR has been observed in younger patients than in older age groups of children; risk is also greater in girls and in patients with a prior history of allergy to other drugs. Management options include cessation of carboplatin and switching to another agent, premedication with antihistamines and/or corticosteroids, and carboplatin desensitization. For sensitized patients who have obtained benefits from carboplatin, the continuation of the treatment is desirable and desensitization protocols have showed promising results. Conclusion: Clinicians must not underestimate the potential risk and occurrence of carboplatin HSRs in the pediatric population in order to outline adequate management strategies. Desensitization protocols should be considered for patients sensitive to carboplatin in order to avoid having to discontinue an effective chemotherapy.


Performance Evaluation of Biozentech Malaria Scanner in Plasmodium knowlesi and P. falciparum as a New Diagnostic Tool.
The computer vision diagnostic approach currently generates several malaria diagnostic tools. It enhances the accessible and straightforward diagnostics that necessary for clinics and health centers in malaria-endemic areas. A new computer malaria diagnostics tool called the malaria scanner was used to investigate living malaria parasites with easy sample preparation, fast and user-friendly. The cultured Plasmodium parasites were used to confirm the sensitivity of this technique then compared to fluorescence-activated cell sorting (FACS) analysis and light microscopic examination. The measured percentage of parasitemia by the malaria scanner revealed higher precision than microscopy and was similar to FACS. The coefficients of variation of this technique were 1.2-6.7% for Plasmodium knowlesi and 0.3-4.8% for P. falciparum. It allowed determining parasitemia levels of 0.1% or higher, with coefficient of variation smaller than 10%. In terms of the precision range of parasitemia, both high and low ranges showed similar precision results. Pearson's correlation test was used to evaluate the correlation data coming from all methods. A strong correlation of measured parasitemia (r2=0.99, P<0.05) was observed between each method. The parasitemia analysis using this new diagnostic tool needs technical improvement, particularly in the differentiation of malaria species.


[Signaling indicator of ventricular fibrillation].
The purpose of the work was to design an appliance for reliable detection of ventricular fibrillation. Frequency-amplitude spectrum of electrocardiograms taken in dogs with normal rhythm and fibrillation, as well as the impulse ratio in normalcy, tachycardia and in fibrillation were investigated. An appliance for automatic separation of ventricular fibrillation by referring to pulse ratio is described. The designed device is shown to have a high degree of discrimination and noise-immunity.


[Functional asymmetry of the cerebral hemispheres in clinical variants of schizophrenia].
The paper is concerned with a study of lefthandedness, the distribution and combinations of characteristics in laterality according to the signs of finger linkage, folding of hands and a directing eye in 610 females and 600 males with schizophrenia. A bimanual correlation of the finger ridges were determined in 560 females and 560 males. In patients of both sexes with continuous schizophrenia there were most frequent cases of lefthandedness, a left type of finger linkage and a left directing eye. Besides, there were more males with a higher ridge on the left hand or equal ridges on both hands. The author believes that the genotypical traits on the organism which are fenotypically expressed in the abovementioned signs exert a modifying influence on the clinical features of schizophrenia.


A novel primer set for multilocus phylogenetic inference in East African cichlid fishes.
The cichlid fishes in the East African Great Lakes are a prime model system for the study of adaptive radiation. Therefore, the availability of an elaborate phylogenetic framework is an important prerequisite. Previous phylogenetic hypotheses on East African cichlids are mainly based on mitochondrial and/or fragment-based markers, and, to date, no taxon-rich phylogeny exists that is based on multilocus DNA sequence data. Here, we present the design of an extensive new primer set (24 nuclear makers) for East African cichlids that will be used for multilocus phylogenetic analyses in the future. The primers are designed to work for both Sanger sequencing and next-generation sequencing with the 454 technology. As a proof of principle, we validate these primers in a phylogenetically representative set of 16 cichlid species from Lake Tanganyika and main river systems in the area and provide a basic evaluation of the markers with respect to marker length and diversity indices.


A liver-selective LXR inverse agonist that suppresses hepatic steatosis.
Fatty liver, which often accompanies obesity and type 2 diabetes, frequently leads to a much more debilitating hepatic disease including non-alcoholic steatohepatitis, cirrhosis, and hepatocellular carcinoma. Current pharmacological therapies lack conclusive efficacy and thus treatment options are limited. Novel therapeutics that suppress either hepatic lipogenesis and/or hepatic inflammation may be useful. Here, we describe the development of the first selective synthetic LXR inverse agonist (SR9238) and demonstrate that this compound effectively suppresses hepatic lipogenesis, inflammation, and hepatic lipid accumulation in a mouse model of non-alcoholic hepatosteatosis. SR9238 displays high potency for both LXRα and LXRβ (40-200 nM IC50) and was designed to display liver specificity so as to avoid potential side effects due to suppression of LXR in the periphery. Unexpectedly, treatment of diet-induced obese mice with SR9238 suppressed plasma cholesterol levels. These data indicate that liver-selective LXR inverse agonists may hold utility in the treatment of liver disease.


An automated solid-phase 17 alpha-hydroxyprogesterone ELISA method using a microtitre plate.
We report an automated ELISA method for the measurement of 17 alpha-hydroxyprogesterone in plasma samples. A rabbit antiserum raised against 17 alpha-hydroxyprogesterone-3-(O-carboxymethyl)oxime-bovine serum albumin is used for the assay and a homologous competitor 17 alpha-hydroxyprogesterone-3-(O-carboxymethyl)oxime-bovine thyroglobulin is coated onto a microtitre plate. A goat anti-rabbit IgG-horse radish peroxidase is used as a probe for this solid-phase assay. The assay exhibits good sensitivity, precision and accuracy. The method is used routinely for the management of patients with congenital adrenal hyperplasia.


Evaluation of Clinical Drug Interaction Potential of Clofazimine Using Static and Dynamic Modeling Approaches.
The 2016 World Health Organization treatment recommendations for drug-resistant tuberculosis (DR-TB) positioned clofazimine as a core second-line drug. Being identified as a cytochrome P450 (P450) inhibitor in vitro, a P450-mediated drug interaction may be likely when clofazimine is coadministered with substrates of these enzymes. The P450-mediated drug interaction potential of clofazimine was evaluated using both static [estimation of the R1 and area under the plasma concentration-time curve ratio (AUCR) values] and dynamic [physiologically based pharmacokinetics (PBPK)] modeling approaches. For static and dynamic predictions, midazolam, repaglinide, and desipramine were used as probe substrates for CYP3A4/5, CYP2C8, and CYP2D6, respectively. The AUCR static model estimations for clofazimine with the substrates midazolam, repaglinide, and desipramine were 5.59, 1.34, and 1.69, respectively. The fold increases in the area under the curve (AUC) predicted for midazolam, repaglinide, and desipramine with clofazimine (based on PBPK modeling) were 2.69, 1.60, and 1.47, respectively. Clofazimine was predicted to be a moderate-to-strong CYP3A4/5 inhibitor and weak CYP2C8 and CYP2D6 inhibitor based on the calculated AUCR by static and PBPK modeling. Additionally, for selected antiretroviral, antitubercular, antihypertensive, antidiabetic, antileprotics, and antihyperlipidemic CYP3A4/5 substrate drugs, approximately 2- to 6-fold increases in the AUC were predicted with static modeling when coadministered with 100 mg of clofazimine. Therefore, the possibility of an increase in the AUC of CYP3A4/5 substrates when coadministered with clofazimine cannot be ignored.


The influence of road curvature on fatal crashes in New Zealand.
Bends in roads can cause crashes but a recent study in the UK found that areas with mostly curved roads had lower crash rates than areas with straighter roads. This present study aimed to replicate the previous research in a different country. Variations in the number of fatal road crashes occurring between 1996 and 2005 in 73 territorial local authorities across New Zealand were modelled against possible predictors. The predictors were traffic flow, population counts and characteristics, car use, socio-economic deprivation, climate, altitude and road characteristics including four measures of average road curvature. The best predictors of the number of fatal crashes on urban roads, rural state highways and other rural roads were traffic flow, speed limitation and socio-economic deprivation. Holding significant factors constant, there was no evidence that TLAs with the most curved roads had more crashes than elsewhere. Fatal crashes on urban roads were significantly and negatively related to two measures of road curvature: the ratio of road length to straight distance and the cumulative angle turned per kilometre. Weaker negative associations on rural state highways could have occurred by chance. These results offer limited support to the suggestion that frequently occurring road bends might be protective.


Flight-muscle polymorphism in the cricket Gryllus firmus: muscle characteristics and their influence on the evolution of flightlessness.
Flight muscles of the cricket Gryllus firmus are polymorphic, existing as pink or white phenotypes. White muscles are smaller in size, have reduced number and size of muscle fibers, and have reduced in vitro enzyme activities and respiration rates relative to pink muscles of newly molted, fully winged adults. G. firmus is also polymorphic for wing length. All newly molted long-winged adults exhibited the pink-muscle phenotype, while most newly molted short-winged adults exhibited the white-muscle phenotype, which resulted from arrested muscle growth. As long-winged adults aged, fully grown pink muscle was transformed into white muscle via histolysis. The substantially higher respiration rate of pink muscle likely contributes to the elevated whole-organism respiration rate of long-winged females, which has been documented previously and which is thought to divert nutrients from egg production. Histolyzed white flight muscle from long-winged crickets also exhibited significantly elevated respiration rate and enzyme activities compared with underdeveloped white muscle from short-winged adults, although these differences were not as great as those between pink and white muscles. Fecundity was much more elevated in females with white verus pink flight muscles than it was in females with short versus long wings. The fitness gain resulting from flightlessness has typically been estimated in previous studies by comparing enhanced egg production of short-winged and long-winged females, without considering the influence of flight-muscle variation. Our results suggest that the magnitude of this fitness gain has been substantially underestimated.


Effects of 3D virtual haptics force feedback on brand personality perception: the mediating role of physical presence in advergames.
This study gauged the effects of force feedback in the Novint Falcon haptics system on the sensory and cognitive dimensions of a virtual test-driving experience. First, in order to explore the effects of tactile stimuli with force feedback on users' sensory experience, feelings of physical presence (the extent to which virtual physical objects are experienced as actual physical objects) were measured after participants used the haptics interface. Second, to evaluate the effects of force feedback on the cognitive dimension of consumers' virtual experience, this study investigated brand personality perception. The experiment utilized the Novint Falcon haptics controller to induce immersive virtual test-driving through tactile stimuli. The author designed a two-group (haptics stimuli with force feedback versus no force feedback) comparison experiment (N = 238) by manipulating the level of force feedback. Users in the force feedback condition were exposed to tactile stimuli involving various force feedback effects (e.g., terrain effects, acceleration, and lateral forces) while test-driving a rally car. In contrast, users in the control condition test-drove the rally car using the Novint Falcon but were not given any force feedback. Results of ANOVAs indicated that (a) users exposed to force feedback felt stronger physical presence than those in the no force feedback condition, and (b) users exposed to haptics stimuli with force feedback perceived the brand personality of the car to be more rugged than those in the control condition. Managerial implications of the study for product trial in the business world are discussed.


Comparison of potassium currents in rabbit atrial and ventricular cells.
1. In rabbit and human hearts there are significant differences in the action potential configuration in atrium and ventricle, and the action potential waveform exhibits marked frequency dependence in both tissues. To study the ionic mechanism(s) of these phenomena, the size and time course of the potassium (K+) currents responsible for repolarization have been recorded from single cells using a whole-cell microelectrode voltage clamp method. 2. At physiological heart rates, the action potential in atrial cells has a short plateau phase; however, the rapid early repolarization is strongly frequency dependent. Ventricular myocytes have a long plateau (400-700 ms at 23 degrees C), and the late repolarizing phase of the action potential is much faster in ventricle than in atrium. 3. In both cell types, four different outward currents can be recorded: (i) a large transient outward current, It; (ii) IK(Ca), a smaller Ca2+-dependent K+ current; (iii) IK, a small, maintained time- and voltage-dependent delayed rectifier K+ current; (iv) IK1, an inwardly rectifying K+ current. 4. It, which is responsible for early repolarization, is much larger in atrium than in ventricle. It has very rapid activation and inactivation kinetics but a very slow time course of recovery from inactivation (tau = 5.4 s at 23 degrees C). Our results show that the reactivation kinetics of It are responsible for the pronounced dependence of the shape of the atrial action potential on stimulus frequency. 5. IK(Ca) is variable from cell to cell and is larger in atrium than in ventricle. In both cell types, IK(Ca) is much smaller than It. 6. The delayed rectifier current, IK, is very small and turns on relatively slowly in both cell types. It is therefore not activated strongly during the relatively short plateau of the atrial action potential. Even in ventricle, it contributes only a small repolarizing current. 7. IK1, the inward rectifier K+ current, is much larger in ventricle than in atrium. The current-voltage relationship for IK1 in ventricle exhibits a negative slope conductance between -50 and 0 mV. IK1 is the outward current which generates the resting membrane potential and it modulates the final repolarization phase of the action potential in both cell types. 8. These data strongly suggest that the action potential configuration and its frequency dependence in rabbit atrial and ventricular cells are mainly due to the differences in sizes and kinetics of It and IK1.


Neurocognitive dysfunction risk alleviation with the use of dexmedetomidine in perioperative conditions or as ICU sedation: a meta-analysis.
Many studies have reported the beneficial effects of dexmedetomidine on postoperative neurocognitive function but overall evidence is not as clear. We examined this conundrum by meta-analyzing studies that used dexmedetomidine in perioperative conditions or as intensive care unit (ICU) sedation and utilized reliable neurocognitive assessment tests. The literature search was undertaken across several electronic databases including EBSCO, Embase, Google Scholar, Ovid SP, PubMed, Scopus, and Web of Science. Literature search was carried out across several electronic databases and relevant studies were selected after following précised inclusion criteria. Meta-analysis of risk differences (RDs) was carried out and subgroup analyses were performed. Twenty studies were selected from which data of 2612 individuals were used. Initial dexmedetomidine dose was 0.68 ± 0.27 and maintenance dose was 0.54 ± 0.32 in the trials. Dexmedetomidine treatment was associated with significantly lower risk of postoperative/postanesthesia neurocognitive dysfunction both in comparison with saline-treated controls (RD [95% confidence interval, CI]: -0.17 (-0.30, -0.04); P = 0.008) and comparators (-0.16 [-0.28, -0.04]; P = 0.009). In the subgroups analyses, however, there was no significant differences between dexmedetomidine and controls/comparators when studies with confusion assessment method for ICU only (RD: -0.10 (-0.22, 0.02); P = 0.1) or midazolam as comparator only (RD: -0.26 (-0.60, 0.07); P = 0.12) were meta-analyzed. Dexmedetomidine use in the perioperative conditions or as ICU sedation is associated with lower risk of neurocognitive dysfunction. There can be some impact of neurocognitive assessment method, drug interactions, and clinical heterogeneity on the overall outcomes of this meta-analysis.


Cadaver donor discards secondary to serology.
The use of cadaveric skin has made a major impact in the survival of patients experiencing major thermal injury. However, the availability of cadaveric skin is often limited by potentially pathogenic organisms. Very little data exists as to why cadaveric skin from donors who have been previously screened was discarded. From March 1994 to March 1996, 813 donors were referred to our tissue bank. All donors were reviewed for the cause of death, history and physical, and social history. One hundred fifty-three donors screened were discarded. Sixty-one donors of this group were discarded because of positive serologies. The following are the percentages of the specific positive serologies: hepatitis B core antibody, 52.3%; hepatitis B surface antigen, 18.1%; hepatitis C virus antibody, 14.3%; human immunodeficiency virus antibody, 4.9%; human T lymphocyte virus antibody, 4.9% and syphilis, 5.5%. Retrospectively, all donor screening questionnaires were reviewed for possible indicators in relation to positive serologic testing. Current screening methods, although excellent in social screening, still fail to identify a significant number of donors who may have positive serologies because of hepatitis, human immunodeficiency virus, human T lymphocyte virus, or syphilis. As the field of tissue banking continues to evolve, the focus will need to be directed toward better screening mechanisms in order to decrease our current discard rates after donors have been approved through the screening process.


Tumor necrosis factor-alpha and interleukin-1alpha decrease the adherence of Streptococcus pyogenes to cultured keratinocytes.
We hypothesized that the primary epidermal cytokines, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1alpha, which are produced after skin injury, modulate bacterial adherence and the initiation of group A streptococcal skin infections. Streptococcus pyogenes binds preferentially to highly differentiated keratinocytes in vitro, simulating the superficial human skin infection, impetigo, and providing a model system for testing this hypothesis. Exposure of keratinocytes to 10 ng/mL TNF-alpha for 20 h decreased adherence to undifferentiated and differentiated keratinocytes by 33% and 38%, respectively. Treatment with 1 ng/mL IL-1alpha decreased adherence to undifferentiated and differentiated keratinocytes by 23% and 18%, respectively. Exposure to both cytokines simultaneously produced an additive 50% reduction in adherence. These data suggest that TNF-alpha and IL-1alpha may play a role in cutaneous host defense by impeding streptococcal adherence and decreasing its ability to form a nidus of infection in the skin.


A suicide attempt by intoxication with Taxus baccata leaves and ultra-fast liquid chromatography-electrospray ionization-tandem mass spectrometry, analysis of patient serum and different plant samples: case report.
Taxus (yew) is one of the most frequently reported plants causing potentially fatal outcome when taken incidentally or for suicidal reasons. A fast and reliable method of detection of poisonous compounds or their metabolites is critical in life-saving procedures in cases of yew ingestion. Previously, several chromatographic analytical procedures have been described usually taking longer than one hour of total analysis time. In this report we describe a suicide case study and an ad hoc developed fast method of detection and quantitation of 3,5-dimethoxyphenol - the main taxane metabolite in the blood plasma from the patient as well as the determination of major taxine components in the plant material (Taxus baccata). At present, there is no reasonable alternative for mass spectrometry that could match its high sensitivity and accuracy, and Multiple Reaction Monitoring could be adequate and useful mass spectrometry technique in analyzing and identification of plants material compounds that cause severe poisoning in humans. In the reported case, intensive cardiac care together with the astuteness of the treating physicians not only saved the patient's life, but also allowed for his complete recovery and return to work. The development of ultra fast liquid chromatography tandem mass spectrometry UFLC-MS/MS method provides a fast means to confirm yew alkaloids and their metabolite in various material. The applied analytical procedure allows early detection of main metabolite in patient material as well as comparing to those extracted from the plant. In our study, the taxanes remained undetected, probably due to the time elapsing from the patient admittance and collection of plasma. In cases like those reported in this study, retaining the gastric material should be obligatory to confirm the ingestion of yew. The possibility of using this approach in detection of native taxine compounds in human plasma remains to be verified.


The self/nonself issue: A confrontation between proteomes.
Defining self and nonself is the most compelling challenge in science today, at the basis of the numerous questions that remain unanswered in the immunology-pathology-therapy debate. The generation of the antibody repertoire, the complicated scenario offered by tolerance and autoimmunity, natural auto-antibodies and their relationship to autoimmune diseases, and positive and negative selection are only a few examples of the unresolved immunological questions. In this context, we proposed that sequence similarity to the host proteome modulates antigen peptide recognition and immunogenicity. Using the available proteome assemblies of viruses, bacteria and higher vertebrates, and applying the low-similarity criterion, we are systematically defining the proteomic similarity of B-cell epitopes already validated experimentally. Here, we report further data documenting that a low similarity to the host proteome is the common property that defines the immunological "nonself" nature of antigenic sequences in cancer, autoimmunity, infectious diseases and allergy.


Expressions of PDGF receptor alpha, c-Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells.
In gastrulating embryos, various types of cells are generated before differentiation into specific lineages. The mesoderm of the gastrulating mouse embryo represents a group of such intermediate cells. PDGF receptor alpha (PDGFRalpha), c-Kit and fetal liver kinase 1 (Flk1) are expressed in distinctive mesodermal derivatives of post-gastrulation embryos. Their expressions during gastrulation were examined by whole mount immunostaining with monoclonal antibodies against these three receptors. The antibodies stained different mesodermal subsets in gastrulating embryos. Flow cytometry of head fold stage embryos revealed that Flk1+ mesodermal cells could be further classified by the level of c-Kit expression. To examine the possibility that hematopoietic cell differentiation is initiated from the Flk1+ mesoderm, embryonic stem (ES) cells were cultured on the OP9 or PA6 stromal cell layer; the former but not the latter supported in vitro hematopoiesis from ES cells. Flk1+ cells were detected only on the OP9 cell layer from day 3 of differentiation before the appearance of hematopoietic cells. Thus, Flk1+ cells will be required for in vitro ES cell differentiation into hematopoietic cells. The results suggest that these three receptor tyrosine kinases will be useful for defining and sorting subsets of mesodermal cells from embryos or in vitro cultured ES cells.


Effects of Mycoplasma spp, Trichomonas fetus, and Campylobacter fetus on ciliary activity of bovine uterine tube organ cultures.
Microscopic observations were made of sections of normal bovine uterine tube (oviduct) and fimbriae maintained in vitro and exposed to agents known to localize in the bovine genital tract, i.e., mycoplasma, Trichomonas fetus, and Campylobacter fetus (Vibrio fetus). Only C fetus stopped ciliary activity. Scanning electron microscopy (SEM) revealed loss of most cilia. Sterile filtrate of C fetus culture had no effect.


Myo-inositol plus selenium supplementation restores euthyroid state in Hashimoto's patients with subclinical hypothyroidism.
Clinical evidence suggests that oral supplementation with myo-inositol (MI) and selenium (Se) is useful in the treatment of autoimmune thyroiditis. The purpose of this study was to highlight the positive response of Hashimoto's patients with subclinical hypothyroidism (SH) treated with MI and Se (MI-Se) in restoring a normal thyroid function. A total of 168 patients with Hashimoto's thyroiditis (HT) having Thyroid Stimulating Hormone (TSH) levels between 3 and 6 µIU/ml were randomized into 2 groups: one receiving MI-Se and the other one Se alone. TSH, anti-thyroid peroxidase (TPOAb) and anti-thyroglobulin (TgAb) levels were significantly decreased in patients treated with combined MI-Se after six months of treatment. Also, a significant free serum T4 increase was observed in MI-Se group, along with an amelioration of patients' quality of life. The administration of MI-Se is significantly effective in decreasing TSH, TPOAb and TgAb levels, as well as in enhancing thyroid hormones and personal wellbeing. Such treatment restored euthyroidism in patients diagnosed with autoimmune thyroiditis.


Subcellular localization of neuronal nitric oxide synthase in the rat nucleus of the solitary tract in relation to vagal afferent inputs.
In the nucleus of the solitary tract (NTS), nitric oxide (NO) modulates neuronal circuits controlling autonomic functions. A proposed source of this NO is via nitric oxide synthase (NOS) present in vagal afferent fibre terminals, which convey visceral afferent information to the NTS. Here, we first determined with electron microscopy that neuronal NOS (nNOS) is present in both presynaptic and postsynaptic structures in the NTS. To examine the relationship of nNOS to vagal afferent fibres the anterograde tracer biotinylated dextran amine was injected into the nodose ganglion and detected in brainstem sections using peroxidase-based methods. nNOS was subsequently visualised using a pre-embedding immunogold procedure. Ultrastructural examination revealed nNOS immunoreactivity in dendrites receiving vagal afferent input. However, although nNOS-immunoreactive terminals were frequently evident in the NTS, none were vagal afferent in origin. Dual immunofluorescence also confirmed lack of co-localisation. Nevertheless, nNOS immunoreactivity was observed in vagal afferent neurone cell bodies of the nodose ganglion. To determine if these labelled cells in the nodose ganglion were indeed vagal afferent neurones nodose ganglion sections were immunostained following application of cholera toxin B subunit to the heart. Whilst some cardiac-innervating neurones were also nNOS immunoreactive, nNOS was never detected in the central terminals of these neurones. These data show that nNOS is present in the NTS in both pre- and postsynaptic structures. However, these presynaptic structures are unlikely to be of vagal afferent origin. The lack of nNOS in vagal afferent terminals in the NTS, yet the presence in some vagal afferent cell bodies, suggests it is selectively targeted to specific regions of the same neurones.


Hiding scattering layers for noninvasive imaging of hidden objects.
The ability to noninvasive image through turbid media has long been a major scientific and technological goal in many disciplines. A breakthrough has been made to observe objects that were completely hidden behind an opaque scattering layer. However, such approach needs not only to scan both illumination light and detector but further off-line procedures to numerically retrieve the image of the objects. Here, we report a distant invisibility-based noninvasive method that can hide scattering layers and allows to directly image objects behind. By recording holograms of the objects through a ground glass and then using the holograms produced time-reversal lights to re-illuminate the objects, we implemented to observe objects with feature size ranging from 39 μm to 80 μm that were hidden behind a 3 mm thick ground glass. Of importance, our approach opens a door towards real-time, high speed biomedical imaging and in-site inspection of integrated devices.


The clinical relevance of self-reported premenstrual worsening of depressive symptoms in the management of depressed outpatients: a STAR*D report.
To determine the incidence, clinical and demographic correlates, and relationship to treatment outcome of self-reported premenstrual exacerbation of depressive symptoms in premenopausal women with major depressive disorder who are receiving antidepressant medication. This post-hoc analysis used clinical trial data from treatment-seeking, premenopausal, adult female outpatients with major depression who were not using hormonal contraceptives. For this report, citalopram was used as the first treatment step. We also used data from the second step in which one of three new medications were used (bupropion-SR [sustained release], venlafaxine-XR [extended release], or sertraline). Treatment-blinded assessors obtained baseline treatment outcomes data. We hypothesized that those with reported premenstrual depressive symptom exacerbation would have more general medical conditions, longer index depressive episodes, lower response or remission rates, and shorter times-to-relapse with citalopram, and that they would have a better outcome with sertraline than with bupropion-SR. At baseline, 66% (n=545/821) of women reported premenstrual exacerbation. They had more general medical conditions, more anxious features, longer index episodes, and shorter times-to-relapse (41.3 to 47.1 weeks, respectively). Response and remission rates to citalopram, however, were unrelated to reported premenstrual exacerbation. Reported premenstrual exacerbation was also unrelated to differential benefit with sertraline and bupropion-SR. Self-reported premenstrual exacerbation has moderate clinical utility in the management of depressed patients, although it is not predictive of overall treatment response. Factors that contribute to a more chronic or relapsing course may also play a role in premenstrual worsening of major depressive disorder (MDD).


Association of plasma Aß peptides with blood pressure in the elderly.
Aß peptides are often considered as catabolic by-products of the amyloid ß protein precursor (APP), with unknown physiological functions. However, several biological properties have been tentatively attributed to these peptides, including a role in vasomotion. We assess whether plasma Aß peptide levels might be associated with systolic and diastolic blood pressure values (SBP and DBP, respectively). Plasma Aß(1-40) and Aß(1-42) levels were measured using an xMAP-based assay in 1,972 individuals (none of whom were taking antihypertensive drugs) from 3 independent studies: the French population-based 3C and MONA-LISA (Lille) studies (n = 627 and n = 769, respectively) and the Australian, longitudinal AIBL study (n = 576). In the combined sample, the Aß(1-42)/ Aß(1-40) ratio was significantly and inversely associated with SBP (p = 0.03) and a similar trend was observed for DBP (p = 0.06). Using the median age (69) as a cut-off, the Aß(1-42)/Aß(1-40) ratio was strongly associated with both SBP and DBP in elderly individuals (p = 0.002 and p = 0.03, respectively). Consistently, a high Aß(1-42)/ Aß(1-40) ratio was associated with a lower risk of hypertension in both the combined whole sample (odds ratio [OR], 0.71; 95% confidence interval [CI], 0.56-0.90) and (to an even greater extent) in the elderly subjects (OR, 0.53; 95% CI, 0.37-0.75). Lastly, all these associations appeared to be primarily driven by the level of plasma Aß(1-40). The plasma Aß(1-42)/Aß(1-40) ratio is inversely associated with SBP, DBP and the risk of hypertension in elderly subjects, suggesting that Aß peptides affect blood pressure in vivo. These results may be particularly relevant in Alzheimer's disease, in which a high Aß(1-42)/Aß(1-40) plasma ratio is reportedly associated with a decreased risk of incident disease.


The Design, Structure-Activity, and kinetic studies of 3-Benzyl-5-oxa-1,2,3,4-Tetrahydro-2H-chromeno-(3,4-c)pyridin-8-yl sulfamates as Steroid sulfatase inhibitors.
Steroid sulfatase inhibitors block the local production of estrogenic steroids and are attractive agents for the treatment of estrogen-dependent cancers. Inspiration of coumarin-based inhibitors, we synthesized thirty-two 5-oxa-1,2,3,4-tetrahydro-2H-chromeno-(3,4-c)pyridin-8-yl sulfamates, focusing on the substitution derivatives on the adjacent phenyl ring and evaluated their abilities to block STS from human placenta and MCF-7 cells. SAR analysis revealed that the incorporation of chlorine at either meta and/or para position of the adjacent phenyl ring of the tricyclic skeleton enhanced STS inhibition. Di-substitutions at the adjacent phenyl ring were superior to mono and tri-substitutions. Further kinetic analysis of these compounds revealed that chloride-bearing compounds, such as 19m, 19v, and 19w, had KI of 0.02 to 0.11 nM and kinact/KI ratios of 8.8-17.5 nM-1min-1, a parameter indicated for the efficiency of irreversible inhibition. We also used the docking model to illustrate the difference in STS inhibitory potency of compounds. Finally, the safety and anti-cancer activity of selected compounds 19m, 19v, and 19w were also studied, showing the results of low cytotoxicity on NHDF cell line and being more potent than irosustat on ZR-75-1 cell, which was a hormone-dependent cancer cell line with high STS expression.