Three high-lysine mutations control the level of ATP-binding HSP70-like proteins in the maize endosperm.
The synthesis and deposition of seed storage proteins in maize are affected by several dominant and recessive mutants. The effect of three independent mutations, floury-2 (fl2), Defective endosperm-B30 (De-B30), and Mucronate (Mc), that reduce zein level in the endosperm were investigated. These mutations also control the level of b-70, a polypeptide bound to protein bodies, which is separable into the two isoforms b-70I and b-70II by two-dimensional gel electrophoresis. Both isoforms are overexpressed 10-fold in fl2; however, only b-70I is present in De-B30 and Mc, which contain an amount of total b-70 isoforms fivefold higher than in the wild type. Both b-70I and b-70II resemble heat shock protein (HSP70) in that they bind ATP, cross-react with anti-HSP antibodies, and have N-terminal sequence homology to HSP70. All maize protein body-located b-70 characteristics are typical of those of chaperone-like HSPs. A third protein, b-70III, similar in size to but slightly more acidic than b-70I and b-70II, also binds ATP and reacts with the same antibody, providing evidence for the presence in endosperm extracts of a cytosolic chaperone-like protein. The level of b-70III was not altered by the mutations studied. The results suggested that the repression effect of the three mutations on zein accumulation may be mediated by the alteration of a zein transport or zein assembly process involving b-70I and b-70II.


Glomus tumor-induced longitudinal splitting of nail mimicking median canaliform dystrophy.
Median canaliform deformity of the nail is an uncommon entity, where there is longitudinal splitting of the nail. Longitudinal splitting of the nail is a rare phenomenon and can also occur following number of growths arising in the nail matrix. On examination there was a longitudinal split in the nail plate, beginning in the distal nail fold and extending proximally all the way to the proximal nail fold. There was a small, almost indiscernible, swelling in that area, which was exquisitely tender. The split part of the nail showed a little discoloration. There was no discharge, bleeding, or subungual mass visible. 'Love test' was positive in this case. After nail avulsion, a small 2 mm x 4 mm nodule was exposed and excised. Histopathological examination of the tumor showed a mantle of glomus cells surrounding the blood vessels.


The Role of ExoS in Dissemination of Pseudomonas aeruginosa during Pneumonia.
Hospital-acquired pneumonia is associated with high rates of morbidity and mortality, and dissemination to the bloodstream is a recognized risk factor for particularly poor outcomes. Yet the mechanism by which bacteria in the lungs gain access to the bloodstream remains poorly understood. In this study, we used a mouse model of Pseudomonas aeruginosa pneumonia to examine this mechanism. P. aeruginosa uses a type III secretion system to deliver effector proteins such as ExoS directly into the cytosol of eukaryotic cells. ExoS, a bi-functional GTPase activating protein (GAP) and ADP-ribosyltransferase (ADPRT), inhibits phagocytosis during pneumonia but has also been linked to a higher incidence of dissemination to the bloodstream. We used a novel imaging methodology to identify ExoS intoxicated cells during pneumonia and found that ExoS is injected into not only leukocytes but also epithelial cells. Phagocytic cells, primarily neutrophils, were targeted for injection with ExoS early during infection, but type I pneumocytes became increasingly injected at later time points. Interestingly, injection of these pneumocytes did not occur randomly but rather in discrete regions, which we designate ""fields of cell injection" (FOCI). These FOCI increased in size as the infection progressed and contained dead type I pneumocytes. Both of these phenotypes were attenuated in infections caused by bacteria secreting ADPRT-deficient ExoS, indicating that FOCI growth and type I pneumocyte death were dependent on the ADPRT activity of ExoS. During the course of infection, increased FOCI size was associated with enhanced disruption of the pulmonary-vascular barrier and increased bacterial dissemination into the blood, both of which were also dependent on the ADPRT activity of ExoS. We conclude that the ADPRT activity of ExoS acts upon type I pneumocytes to disrupt the pulmonary-vascular barrier during P. aeruginosa pneumonia, leading to bacterial dissemination.


An innate immune response and altered nuclear receptor activation defines the spinal cord transcriptome during alpha-tocopherol deficiency in Ttpa-null mice.
Mice with deficiency in tocopherol (alpha) transfer protein gene develop peripheral tocopherol deficiency and sensory neurodegeneration. Ttpa-/- mice maintained on diets with deficient α-tocopherol (α-TOH) had proprioceptive deficits by six months of age, axonal degeneration and neuronal chromatolysis within the dorsal column of the spinal cord and its projections into the medulla. Transmission electron microscopy revealed degeneration of dorsal column axons. We addressed the potential pathomechanism of α-TOH deficient neurodegeneration by global transcriptome sequencing within the spinal cord and cerebellum. RNA-sequencing of the spinal cord in Ttpa-/- mice revealed upregulation of genes associated with the innate immune response, indicating a molecular signature of microglial activation as a result of tocopherol deficiency. For the first time, low level Ttpa expression was identified in the murine spinal cord. Further, the transcription factor liver X receptor (LXR) was strongly activated by α-TOH deficiency, triggering dysregulation of cholesterol biosynthesis. The aberrant activation of transcription factor LXR suppressed the normal induction of the transcription factor retinoic-related orphan receptor-α (RORA), which is required for neural homeostasis. Thus we find that α-TOH deficiency induces LXR, which may lead to a molecular signature of microglial activation and contribute to sensory neurodegeneration.


Hot-SWV: Square Wave Voltammetry with Hot Microelectrodes.
A promising strategy to lowering detection limits in electrochemical analysis is the active modulation of the electrode temperature. Specifically, by tuning the electrode's surface temperature one can enhance detection limits due to improved electrode process kinetics and increased mass transfer rates, all without affecting the bulk solution. Motivated by this argument, here we report the development of a new electroanalytical technique based on electrode-temperature modulation, which we call hot square wave voltammetry (Hot-SWV). The technique utilizes the superposition of conventional SWV, already considered as one of the most sensitive voltammetric techniques, and a high frequency alternating current (ac) waveform to electrically polarize microelectrodes. By applying about 100 MHz ac frequencies (with varying Vrms amplitudes), our method generates an electrothermal fluid flow (ETF) in the electrolyte surrounding the electrode, thereby increasing the sensitivity of the SWV-based detection. We demonstrate this by investigating the oxidation of ferrocyanide and iron(II) ions, as well as the reduction of the coordination compound ruthenium(III) hexamine under various experimental conditions. We validate our experimental results against a theoretical model built using finite element analysis and observe agreement within ≤15% error at temperatures ≤39 °C. Using Hot-SWV, we observe at least one-order-of-magnitude improvement in the limit of detection of ferrocyanide ions relative to conventional, mm-size electrodes at 25 °C. In addition, we anticipate that Hot-SWV will be particularly useful for electroanalytical measurements of ultralow (≤pM) concentrations of analytes in environmental and biomedical applications.


Electron microscopic morphometry.
The fundamental concepts of morphometry, the principal correction factors for systematic errors and the basic principles of efficient sampling are outlined for quantitative morphology at the electron microscopic level of resolution. The important usefulness of correlating electron microscopic morphometry with complementary light microscopic morphometry is emphasized.


Squaraine dyes in PDT: from basic design to in vivo demonstration.
The design and development of novel squaraine dyes as sensitisers for photodynamic therapy (PDT) applications has grown tremendously in the last decade from the time when a squaraine dye was proposed to be a potential candidate, to-date when the use of such dyes have been demonstrated in animal models for skin cancer. This perspective article highlights the basic design, tuning of absorption, triplet excited state and two-photon absorption properties and recent developments of the squaraines as PDT sensitisers.


Heart failure after myocardial infarction: prevalence of preserved left ventricular systolic function in the community.
Studies have reported that a large proportion of the cases of congestive heart failure (CHF) with mixed etiologies have preserved left ventricular systolic function. Whether this is the case in subjects with CHF after myocardial infarction (MI) is not known. This study was undertaken to examine the prevalence and characteristics associated with CHF in patients who had preserved ejection fraction (LVEF) after MI. Clinical characteristics and LVEF were ascertained in a population-based cohort of patients with CHF after incident MI in Olmsted County, Minn. All MIs were validated by use of standardized epidemiological criteria, and all episodes of CHF were validated by use of Framingham criteria. Between 1979 and 1994, 1658 patients had an MI, and 644 of these patients (38%) had CHF during 7.4 +/- 5.4 years of follow-up. Of these patients, 395 (61%) underwent LVEF assessment. Preserved LVEF (ie, > or =50%) was present in 30% of cases, and this proportion did not change with time. The proportion of women with CHF and preserved LVEF (37%) was greater than the proportion of men (23%, P =.002). The positive association between female sex and preserved LVEF remained significant after adjustment (odds ratio 1.97, 95% CI 1.26-3.07, P =.003). The highest tertile of peak creatinine phosphokinase level was negatively associated with preserved LVEF (odds ratio 0.51, 95% CI 0.29-0.89). A notable proportion of cases of CHF after MI have preserved LVEF. This underscores the burden of CHF with preserved LVEF in a well-defined group of patients with documented coronary disease. CHF with preserved LVEF after MI is associated with female sex and smaller MI size.


An estimate of the rate at which crocidolite asbestos fibres are cleared from the lung.
The cumulative fibre exposure of 36 deceased persons who had been employed on crocidolite asbestos mines of the NW Cape was compared with their lung burden determined by fibre count by optical microscope. Allowance for the period which lapsed between date of last exposure and death revealed a lung clearance rate amounting to a half-life of about 6 years.


Detection of pathogenic protozoa in the diagnostic laboratory: result reproducibility, specimen pooling, and competency assessment.
Stool microscopy as performed in clinical parasitology laboratories is a complex procedure with subjective interpretation. Quality assurance (QA) programs often emphasize proficiency testing as an assessment tool. We describe a result reproducibility assessment tool, which can form part of a broader QA program, and which is based on the blinded resubmission of selected clinical samples, using concordance between the reports of the initial and resubmitted specimen as an indicator. Specimens preserved in sodium acetate-acetic acid-formalin can be stored for several months for use in such a program. The presence of multiple protozoa in one specimen does not affect concordance. Some dilution of specimens occurs in this process, and this may explain poor concordance when specimens with low protozoal concentrations are resubmitted. Evaluation of this tool in a large parasitology laboratory revealed concordance rates for pathogenic protozoa (Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Dientamoeba fragilis) of about 80%, which may be considered for use as a benchmark value. We also used this tool to demonstrate that when pairs of specimens from one patient are pooled to create a single specimen, concordance between the results of the individual and pooled specimens is high.


[The Child and Adolescent Health Survey: collection of health behavior data from parents and children].
The first German Health Survey for Children and Adolescents will start in 2003. During three years, a representative sample of children and adolescents between 0 and 18 years of age shall be examined and will, together with their parents, be interviewed on health-relevant issues. With this survey, health information on about 18,000 children will be gathered. The health behaviour of the parents as far as their children's health is concerned, as well as the personal health behaviour during adolescence will be assessed, since this will be highly relevant for future health. Questions on nutrition behaviour, eating disorders, alcohol consumption, smoking as well as leisure-time behaviour, with special emphasis on physical activity, will be asked. These questions were tested for feasibility in a one-year pilot study. With this survey, the occurrence of certain unhealthy behaviour characteristics may be documented and risk groups may be identified. This may help to conceptualize prevention measures.


Hemodynamic effects of nitrous oxide in isoflurane-anesthetized cats.
To determine the hemodynamic effects of nitrous oxide in isoflurane-anesthetized cats. 12 healthy adult domestic shorthair cats. Cats were anesthetized by administration of isoflurane in oxygen. After instruments were inserted, end-tidal isoflurane concentration was set at 1.25 times the individual minimum alveolar concentration, and nitrous oxide was administered in a Latin-square design at 0, 30, 50, and 70%. Each concentration was administered for 25 minutes before measurements were obtained to allow for stabilization. Heart rate; systemic and pulmonary arterial pressures; central venous pressure; pulmonary artery occlusion pressure; cardiac output; body temperature; arterial and mixed-venous pH, PCO2, PO2, and hemoglobin concentrations; PCV; and total protein and lactate concentrations were measured before and during noxious stimulation for each nitrous oxide concentration. Arterial and mixed-venous bicarbonate concentrations and oxygen saturation, cardiac index, stroke index, rate-pressure product, systemic and pulmonary vascular resistance indices, left and right ventricular stroke work indices, arterial and mixed-venous oxygen contents, oxygen delivery, oxygen consumption, oxygen extraction ratio, alveolar-to-arterial oxygen difference, and venous admixture were calculated. Arterial pressure, central venous pressure, pulmonary arterial pressure, rate-pressure product, systemic and pulmonary vascular resistance indices, arterial PCO2, and PCV increased during administration of 70% nitrous oxide. Arterial and mixed-venous pH, mixed-venous PO2, and alveolar-to-arterial oxygen difference decreased during administration of 70% nitrous oxide. Results before and during noxious stimulation were similar. Administration of 70% nitrous oxide to isoflurane-anesthetized cats resulted in improved arterial pressure, which was related to a vasoconstrictive effect.


Ascitic fluid analysis in peritoneal carcinomatosis: comparison of various biochemical tests with ascitic cirrhotics.
Owing to the high false-negative rate of cytological examinations, various biochemical tests have been investigated to search for an alternative method to differentiate between the most frequent causes of ascites formation, i.e. cirrhosis of the liver and peritoneal carcinomatosis. For this purpose, serum and ascitic fluid samples of 71 ascitic patients (54 due to cirrhosis of the liver and 17 due to peritoneal carcinomatosis) were prospectively and simultaneously collected. Eleven major parameters were analyzed, and, of them, ascitic fluid protein, albumin, cholesterol, fibronectin and serum-ascitic gradient of protein and albumin were found to be useful. The serum-ascitic fluid albumin gradient was not as useful as described in the literature and, if clinically applied, the best cut-off value should be raised up to 1.5 gm/dl. Neither as useful was the serum-ascitic fluid protein gradient. Ascitic cholesterol and ascitic fibronectin were more accurate than the ascitic total protein, ascitic albumin and serum-ascitic gradient of protein and albumin in the diagnosis of peritoneal carcinomatosis. Of these tests, an ascitic fluid fibronectin concentration greater than 40 micrograms/ml was the most accurate parameter (diagnostic accuracy 96.2%) that could solely help to establish the diagnosis of peritoneal carcinomatosis.


Ghrelin-containing neurons in the olfactory bulb send collateralized projections into medial amygdaloid and arcuate hypothalamic nuclei: neuroanatomical study.
Ghrelin, a gastrointestinal hormone, is a modulator of the sense of smell. The main source of ghrelin in the central nervous system has been mainly observed in specific populations of hypothalamic neurons. An increasing number of studies have reported ghrelin synthesis and its effect on neurons outside the hypothalamus. Ghrelin and its receptors are expressed in the olfactory bulbs and in other centres of the brain, such as the amygdala, for processing olfactory signals, pyramidal neurons of the cerebral cortex and the dorsal vagal complex of the medulla oblongata. It is known that ghrelin is involved in cognitive mechanisms and eating behaviours, in fact, its expression increases in anticipation of food intake. In order to identify the existence of centrifugal direct afferents from the main olfactory bulb to the medial amygdala and the hypothalamus arcuate nucleus, in this work we used two retrograde tracers, Dil and Fluoro Gold, and immunohistochemical procedure to visualize positive ghrelin neurons. Our paper provides neuroanatomic support for the ghrelin modulation of smell. Our results show that ghrelin neuron projections from mitral cells of bulbs can transmit olfactory information via branching connections to the amygdala and the hypothalamus. This pathway could play an important role in regulating feeding behaviour in response to odours.


Calmer: a robot for managing acute pain effectively in preterm infants in the neonatal intensive care unit.
For preterm infants in the neonatal intensive care unit, early exposure to repeated procedural pain is associated with negative effects on the brain. Skin-to-skin contact with parents has pain-mitigating properties, but parents may not always be available during procedures. Calmer, a robotic device that simulates key pain-reducing components of skin-to-skin contact, including heart beat sounds, breathing motion, and touch, was developed to augment clinical pain management. Our objective was to evaluate the initial efficacy of Calmer for mitigating pain in preterm infants. We hypothesized that, compared to babies who received a human touch-based treatment, facilitated tucking, infants on Calmer would have lower behavioural and physiological pain indices during a single blood test required for clinical care. Forty-nine preterm infants, born between 27 and 36 weeks of gestational age, were randomized either to facilitated tucking or Calmer treatment. Differences between groups in changes across 4 procedure phases (baseline 1, baseline 2, poke, and recovery) were evaluated using (1) the Behavioral Indicators of Infant Pain scored by blind coders from bedside videotape and (2) heart rate and heart rate variability continuously recorded from a single-lead surface ECG (lead II) (Biopac, Canada) sampled at 1000 Hz using a specially adapted portable computer system and processed using Mindware. No significant differences were found between groups on any outcome measures. Calmer provided similar treatment efficacy to a human touch-based treatment. More research is needed to determine effects of Calmer for stress reduction in preterm infants in the neonatal intensive care unit over longer periods.


A review of continuation electroconvulsive therapy: application, safety, and efficacy.
Electroconvulsive therapy (ECT) is a neurostimulation therapeutic intervention that is highly effective and frequently used to treat certain psychiatric conditions, particularly major depressive disorder. Despite its high efficacy, a major limitation of ECT is the significant rate at which patients relapse after treatment. Providing additional ECT treatments after completion of a short-term course of ECT, referred to as continuation ECT (C-ECT), is a strategy used to reduce the risk of relapse. Specifically, C-ECT involves the administration of additional ECT treatments during the 6-month period after remission. This article summarizes the available literature regarding C-ECT including indication for use, patient selection, treatment guidelines/parameters, and safety. The efficacy of C-ECT is also discussed, with a focus on major depressive disorder and schizophrenia. On the basis of the current literature, indications for use and patient selection for C-ECT are predominately similar to those for a short-term ECT course. The treatment guidelines/parameters for C-ECT are recommended to be consistent with the parameters used to achieve remission, with the exception of greater intertreatment intervals during C-ECT. Although adverse cognitive effects can occur during C-ECT, the risk and severity of cognitive impairment are generally low, possibly because of the greater intertreatment intervals. Most research supports the use of C-ECT to prolong remission; however, methodologic limitations mitigate firm conclusions and generalizability of these findings. Nonetheless, the available evidence supports the use of C-ECT as a safe and effective method in relapse prevention.


[Vitamin D3 availability and functional activity of peripheral blood phagocytes in experimental type 1 diabetes].
The study was devoted to identifying the relation between vitamin D3 availability (assessed by the level of circulatory 25OHD3), content of vitamin D3 25-hydroxylase isozymes CYP27A1 and CYP2R1 in hepatic tissue and functional activity of peripheral blood phagocytes in mice with experimental type 1 diabetes. It has been shown that diabetes is accompanied by the development of vitamin D3-deficiency which is characterized by decreased 25OHD3 content in blood serum and determined by changes in tissue expression of the major isoforms of vitamin D3 25-hydroxylase. The level of hepatic CYP27A1 was revealed to be markedly reduced with a concurrent significant augmentation of CYP2R1. Cholecalciferol administration resulted in normalization of tissue levels of both isoforms of vitamin D3 25-hydroxylase and blood serum 25OHD3 content. Diabetes-associated vitamin D3 deficiency correlated with a decrease in phagocytic activity of granulocytes and monocytes, and their ability to produce antibacterial biooxidants such as reactive oxygen and nitrogen forms. Vitamin D3 efficacy to attenuate these abnormalities of immune function was established, indicating an important immunoregulatory role of cholecalciferol in the phagocytic mechanism of antigens elimination implemented by granulocytes and monocytes.


Juror reactions to jury duty: perceptions of the system and potential stressors.
Jurors were surveyed on their general perceptions of the court system and factors that may cause stress immediately after trial, after participating in a post-trial debriefing, and a month after trial. Jurors had an overall positive view of the court system but did report some perceived inequities. The two most stressful elements of jury duty were related to the complexity of the trial and the decision-making involved in the trial, although jurors reported low levels of stress overall. Women reported more stress than men, and trial characteristics such as trial length also affected stress levels. The debriefing intervention was perceived as helpful, but jurors' stress levels were similar at pre- and post-debriefing. Finally, although stress on some measures was lower at the 1 month follow-up, this reduction was not moderated by whether or not jurors received the debriefing.


[Anterior thoracic transdural spinal cord herniation: a sporadic, treatable cause of progressive myelopathy].
Anterior thoracic transdural spinal cord herniation (ATTSCH) is a rare cause of slowly progressing thoracic myelopathy. The myelopathy is caused by herniation and subsequent trapping of the thoracic spinal cord through a defect in the anterior dural sac. We describe the case of a man, 38 years of age, who presented with a 4-year history of progressive pyramidal syndrome with sphincteral and erection disorders caused by ATTSCH. The patient underwent a surgical intradural exploration and release (detethering) of the spinal cord and reduction of the cord herniation; a polytetrafluoroethylene sleeve was inserted to cover the defect and to prevent the cord from reherniation. Anterior thoracic transdural spinal cord herniation is a treatable cause of myelopathy and early detection is of great importance to the prognosis. Because this condition is so rare and therefore, relatively unknown, delay in the diagnostic process and subsequent surgical treatment often occur.


Evaluation of easily measured risk factors in the prediction of osteoporotic fractures.
Fracture represents the single most important clinical event in patients with osteoporosis, yet remains under-predicted. As few premonitory symptoms for fracture exist, it is of critical importance that physicians effectively and efficiently identify individuals at increased fracture risk. Of 3426 postmenopausal women in CANDOO, 40, 158, 99, and 64 women developed a new hip, vertebral, wrist or rib fracture, respectively. Seven easily measured risk factors predictive of fracture in research trials were examined in clinical practice including: age (< 65, 65-69, 70-74, 75-79, 80+ years), rising from a chair with arms (yes, no), weight (< 57, > or = 57 kg), maternal history of hip fracture (yes, no), prior fracture after age 50 (yes, no), hip T-score (> -1, -1 to > -2.5, < or = -2.5), and current smoking status (yes, no). Multivariable logistic regression analysis was conducted. The inability to rise from a chair without the use of arms (3.58; 95% CI: 1.17, 10.93) was the most significant risk factor for new hip fracture. Notable risk factors for predicting new vertebral fractures were: low body weight (1.57; 95% CI: 1.04, 2.37), current smoking (1.95; 95% CI: 1.20, 3.18) and age between 75-79 years (1.96; 95% CI: 1.10, 3.51). New wrist fractures were significantly identified by low body weight (1.71, 95% CI: 1.01, 2.90) and prior fracture after 50 years (1.96; 95% CI: 1.19, 3.22). Predictors of new rib fractures include a maternal history of a hip fracture (2.89; 95% CI: 1.04, 8.08) and a prior fracture after 50 years (2.16; 95% CI: 1.20, 3.87). This study has shown that there exists a variety of predictors of future fracture, besides BMD, that can be easily assessed by a physician. The significance of each variable depends on the site of incident fracture. Of greatest interest is that an inability to rise from a chair is perhaps the most readily identifiable significant risk factor for hip fracture and can be easily incorporated into routine clinical practice.


In-silico study of drug delivery to atherosclerosis in the human carotid artery using metal-organic frameworks based on adhesion of nanocarriers.
This study investigates nanocarriers (NCs) for drug delivery targeting carotid artery atherosclerosis. This targeted drug delivery mechanism is based on ligand-receptor bindings facilitated by coating NCs with P-selectin aptamers, which exhibit high affinities for P-selectin plaque receptors. Recognizing the significant advantages of metal-organic frameworks (MOFs), such as their high drug-loading percentages, we chose them as nanocarriers for this research. Our evaluation considers critical factors: NC surface density (the number of attached nanocarriers per unit of plaque area), toxicity (percentage of NCs missing the target), and efficient drug transfer to plaque tissue. Employing molecular dynamics (MD) for drug loading calculations via van der Waals interactions and computational fluid dynamics (CFD) for toxicity, surface density, and drug transfer assessments, we achieve a comprehensive analysis. A cardiac cycle-based metric guides optimal MOF release conditions, establishing an ideal dosage of 600 NCs per cycle. MOF-801 exhibits outstanding drug delivery performance, particularly in plaque targeting. While a magnetic field enhances NC adhesion, its impact on drug transfer is limited, emphasizing the need for further optimization in magnetic targeting for NC-based therapies. This study provides crucial insights into NC drug delivery performance in carotid artery atherosclerosis, advancing the field of targeted drug delivery for atherosclerosis treatment.


Mucin degradation in human colon ecosystems. Degradation of hog gastric mucin by fecal extracts and fecal cultures.
The enteric flora of rats plays a major role in the degradation of their gut mucin glycoproteins, but the role of human enteric flora in degrading gut mucin glycoprotein has not been as well defined. We have studied the degradation of hog gastric mucin, which is structurally similar to human gastric mucin, in anaerobic human fecal cultures and by partially purified enzyme fractions from human fecal extracts and anaerobic culture supernates. Extensive degradation of the mucin carbohydrate moieties occurred in all systems, but the degradation of mucin protein was less. During a 48-h incubation, the average percent degradation of mucin carbohydrates and mucin protein were respectively 96% and 57% in eight fecal cultures, 66% and 15% by four fecal culture supernates, and 78% and 43% by three fecal extracts. Degradation of mucin protein was greater during bacterial growth in fecal cultures than during incubation with fecal culture supernates, suggesting that bacterial protease activity was predominantly cell bound. We conclude that, as in the rat, the human enteric microflora degrades mucin carbohydrate moieties extensively and the mucin protein to a lesser extent.


[Determination of the affinity of high- and low-affinity antibodies, which are in a mixture, using ELISA and the method of non-linear regression].
It was shown that application of the method of non-linear regression for the solution of the equation, which relates the fraction of free antibodies in a mixture and antigen concentrations, allows to determine the affinity constants for two antibodies in a mixture. Such method is easier and more accurate than the suggested by us earlier method, which use the numerical solution of the appropriate four equations, that describe the relations between the experimental data obtained by ELISA, competing antigen concentration, and values of antibody affinity. In addition, the proposed method allows using much less quantity of experimental measurements without diminishing of the accuracy for the affinity constants evaluations.


Effectiveness of polypill for primary and secondary prevention of cardiovascular diseases (PolyIran): a pragmatic, cluster-randomised trial.
A fixed-dose combination therapy (polypill strategy) has been proposed as an approach to reduce the burden of cardiovascular disease, especially in low-income and middle-income countries (LMICs). The PolyIran study aimed to assess the effectiveness and safety of a four-component polypill including aspirin, atorvastatin, hydrochlorothiazide, and either enalapril or valsartan for primary and secondary prevention of cardiovascular disease. The PolyIran study was a two-group, pragmatic, cluster-randomised trial nested within the Golestan Cohort Study (GCS), a cohort study with 50 045 participants aged 40-75 years from the Golestan province in Iran. Clusters (villages) were randomly allocated (1:1) to either a package of non-pharmacological preventive interventions alone (minimal care group) or together with a once-daily polypill tablet (polypill group). Randomisation was stratified by three districts (Gonbad, Aq-Qala, and Kalaleh), with the village as the unit of randomisation. We used a balanced randomisation algorithm, considering block sizes of 20 and balancing for cluster size or natural log of the cluster size (depending on the skewness within strata). Randomisation was done at a fixed point in time (Jan 18, 2011) by statisticians at the University of Birmingham (Birmingham, UK), independent of the local study team. The non-pharmacological preventive interventions (including educational training about healthy lifestyle-eg, healthy diet with low salt, sugar, and fat content, exercise, weight control, and abstinence from smoking and opium) were delivered by the PolyIran field visit team at months 3 and 6, and then every 6 months thereafter. Two formulations of polypill tablet were used in this study. Participants were first prescribed polypill one (hydrochlorothiazide 12·5 mg, aspirin 81 mg, atorvastatin 20 mg, and enalapril 5 mg). Participants who developed cough during follow-up were switched by a trained study physician to polypill two, which included valsartan 40 mg instead of enalapril 5 mg. Participants were followed up for 60 months. The primary outcome-occurrence of major cardiovascular events (including hospitalisation for acute coronary syndrome, fatal myocardial infarction, sudden death, heart failure, coronary artery revascularisation procedures, and non-fatal and fatal stroke)-was centrally assessed by the GCS follow-up team, who were masked to allocation status. We did intention-to-treat analyses by including all participants who met eligibility criteria in the two study groups. The trial was registered with ClinicalTrials.gov, number NCT01271985. Between Feb 22, 2011, and April 15, 2013, we enrolled 6838 individuals into the study-3417 (in 116 clusters) in the minimal care group and 3421 (in 120 clusters) in the polypill group. 1761 (51·5%) of 3421 participants in the polypill group were women, as were 1679 (49·1%) of 3417 participants in the minimal care group. Median adherence to polypill tablets was 80·5% (IQR 48·5-92·2). During follow-up, 301 (8·8%) of 3417 participants in the minimal care group had major cardiovascular events compared with 202 (5·9%) of 3421 participants in the polypill group (adjusted hazard ratio [HR] 0·66, 95% CI 0·55-0·80). We found no statistically significant interaction with the presence (HR 0·61, 95% CI 0·49-0·75) or absence of pre-existing cardiovascular disease (0·80; 0·51-1·12; pinteraction=0·19). When restricted to participants in the polypill group with high adherence, the reduction in the risk of major cardiovascular events was even greater compared with the minimal care group (adjusted HR 0·43, 95% CI 0·33-0·55). The frequency of adverse events was similar between the two study groups. 21 intracranial haemorrhages were reported during the 5 years of follow-up-ten participants in the polypill group and 11 participants in the minimal care group. There were 13 physician-confirmed diagnoses of upper gastrointestinal bleeding in the polypill group and nine in the minimal care group. Use of polypill was effective in preventing major cardiovascular events. Medication adherence was high and adverse event numbers were low. The polypill strategy could be considered as an additional effective component in controlling cardiovascular diseases, especially in LMICs. Tehran University of Medical Sciences, Barakat Foundation, and Alborz Darou.


Nuclear factor erythroid 2-related factor 2 facilitates neuronal glutathione synthesis by upregulating neuronal excitatory amino acid transporter 3 expression.
Astrocytes support neuronal antioxidant capacity by releasing glutathione, which is cleaved to cysteine in brain extracellular space. Free cysteine is then taken up by neurons through excitatory amino acid transporter 3 [EAAT3; also termed Slc1a1 (solute carrier family 1 member 1)] to support de novo glutathione synthesis. Activation of the nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant responsive element (ARE) pathway by oxidative stress promotes astrocyte release of glutathione, but it remains unknown how this release is coupled to neuronal glutathione synthesis. Here we evaluated transcriptional regulation of the neuronal cysteine transporter EAAT3 by the Nrf2-ARE pathway. Nrf2 activators and Nrf2 overexpression both produced EAAT3 transcriptional activation in C6 cells. A conserved ARE-related sequence was found in the EAAT3 promoter of several mammalian species. This ARE-related sequence was bound by Nrf2 in mouse neurons in vivo as observed by chromatin immunoprecipitation. Chemical activation of the Nrf2-ARE pathway in mouse brain increased both neuronal EAAT3 levels and neuronal glutathione content, and these effects were abrogated in mice genetically deficient in either Nrf2 or EAAT3. Selective overexpression of Nrf2 in brain neurons by lentiviral gene transfer was sufficient to upregulate both neuronal EAAT3 protein and glutathione content. These findings identify a mechanism whereby Nrf2 activation can coordinate astrocyte glutathione release with neuronal glutathione synthesis through transcriptional upregulation of neuronal EAAT3 expression.


Response to Comments on "Outburst flood at 1920 BCE supports historicity of China's Great Flood and the Xia dynasty".
Wu et al, Han, and Huang et al question our reconstruction of a large outburst flood and its possible relationship to China's Great Flood and the Xia dynasty. Here, we clarify misconceptions concerning geologic evidence of the flood, its timing and magnitude, and the complex social-cultural response. We also further discuss how this flood may be related to ancient accounts of the Great Flood and origins of the Xia dynasty.


A Linked Community and Health Facility Intervention to Improve Newborn Health in Cambodia: the NICCI Stepped-Wedge Cluster-Randomized Controlled Trial.
Newborn mortality in Cambodia remains high, with sepsis and complications of delayed care-seeking important contributing factors. Intervention study objectives were to improve infection control behavior by staff in health centers; improve referral of sick newborns; increase recognition of danger signs, and prompt care-seeking at an appropriate health facility; and appropriate referral for sick newborns by mothers and families of newborn infants. The stepped-wedge cluster-randomized controlled trial took place in rural Cambodia from February 2015 to November 2016. Sixteen clusters consisted of public health center catchment areas serving the community. The intervention included health center staff training and home visits to mothers by community health volunteers within 24 hours of birth and on days 3 and 7 after delivery, including assessment of newborns for danger signs and counselling mothers. The trial participants included women who had recently delivered a newborn who were visited in their homes in the first week, as well as health center staff and community volunteers who were trained in newborn care. Women in their last trimester of pregnancy greater than 18 years of age were recruited and were blinded to their group assignment. Mothers and caregivers (2494) received counseling on handwashing practices, breastfeeding, newborn danger signs, and prompt, appropriate referral to facilities. Health center staff in the intervention group had increased likelihood of hand washing at recommended key moments when compared with the control group, increased knowledge of danger signs, and higher recall of at least three hygiene messages. Of mother/caregiver participants at 14 days after delivery, women in the intervention group were much more likely to know at least three danger signs and to have received messages on care-seeking compared with controls. The intervention improved factors understood to be associated with newborn survival and health. Well-designed training, followed by regular supervision, enhanced the knowledge and self-reported behavior of health staff and health volunteers, as well as mothers' own knowledge of newborn danger signs. However, further improvement in newborn care, including care-seeking for illness and handwashing among mothers and families, will require additional involvement from broader stakeholders in the community.


Vascular reactivity in young adults and cardiovascular disease. A prospective study.
Cardiovascular reactivity in response to the cold pressor test has been associated with an increased risk of coronary heart disease in middle-aged men. We studied 905 white male medical students, median age 22 years, in the Johns Hopkins Precursors Study. Systolic blood pressure, systolic blood pressure change during the cold pressor test, smoking, cholesterol, Quetelet index, and family history of coronary heart disease were measured on enrollment during 1948-1964. Incidence of cardiovascular morbidity and mortality was ascertained by annual questionnaires and death certificates. There was no association between change in systolic blood pressure during the cold pressor test, whether examined as a continuous variable or a 20 mm Hg or more rise, and the risk of subsequent cardiovascular disease or coronary heart disease. These findings did not change after adjustment for cardiovascular disease risk factors. Previously reported associations may have been due to preexisting arteriosclerosis, which increases the rise in systolic blood pressure during the cold pressor test. We conclude that cardiovascular reactivity to the cold pressor test in young adulthood is not a strong predictor of future cardiovascular disease.


[Effects of methylflavonolamine hydrochloride on action potentials of isolated guinea pig papillary muscles].
Methylflavonolamine hydrochloride (4'-methyl-7-(2-hydroxy-3-isopropylamino-propoxy)-flavone hydrochloride, MFA) is a new compound synthesized by Shanghai Institute of Pharmaceutical Industry in China. Its effects on the fast and evoked slow response action potentials of isolated guinea pig papillary muscles have been studied by intracellular microelectrode techniques. The data were automatically analysed by a microcomputer system. At concentrations of 5-75 mumol/L, MFA caused a significant decrease in the duration required for 30, 50, 90% repolarization and a shift in the plateau to more negative transmembrane potentials. MFA 5 mumol/L had no effect on the maximal upstroke velocity during phase 0 (Vmax) but decreased it at 75 mumol/L. MFA had no significant effects on other AP parameters. It significantly inhibited the slow AP induced by isoprenaline or histamine (1 mumol/L) after myocardial cells were depolarized in high K+ (25 mmol/L) solution. The results suggest that MFA inhibits the inward Ca2+ current and at high concentration inhibits also the Na+ current.


An acoustically evoked short latency negative response in profound hearing loss patients.
In our auditory brainstem response (ABR) tests, a peculiar V-shaped negative wave at around 3 4 ms latency was observed. At present, this acoustically evoked short latency negative response (ASNR) is poorly understood. In order to study its appearance and origin, the ABR data of 3104 tests during 1980-98 were reviewed. The ASNR was found only in profound hearing loss ears under intense stimuli (80-120 dB nHL). Out of the total 653 profound hearing loss patients (981 ears), the click-evoked ASNRs were present in 80 patients (12.3%), 117 ears (11.9%). The age range was from 8 months to 70 years. Demographic outcome revealed significant higher appearance rates in young subjects especially in the 20-30 years group. The ASNR was excluded from an artifact by its reproducibility over time, equipment and institutes. Moreover, it became absent after external auditory canal occlusion, which simply blocked the air conduction without any influence upon scalp potentials or equipment. It had neural response characteristics that the latency and amplitude shortened and increased respectively in response to the increase of stimulus intensity. Because the peculiar V-shaped waveform obviously differs from ABR, the ASNR was not interpreted as a potential generated from the conventional auditory pathway. On the other hand, the ASNR individuals were of good vestibular function in sharp contrast with their poor hearing. This suggests the probable relation between the ASNR and the vestibular system. The saccule and vestibular nucleus are hypothesized to be the sense organ and the origin of the response respectively.


A dynamic Web application within an n-tier architecture: a Multi-Source Information System for end-stage renal disease.
A Multi-Source Information System (MSIS) has been designed for the Renal Epidemiology and Information Network (REIN) dedicated to End-Stage Renal Disease. Interoperability has been considered at 4 levels: semantics, network, formats and contents. An n-tier architecture has been chosen at the network level. It is made out of a universal client, a dynamic Web server connected to a production database and to a data warehouse. The MSIS is patient-oriented, based on a regional organization. Its implementation in the context of a regional experimentation is presented with insights on the design and underlying technologies. The n-tier architecture is a robust model and flexible enough to aggregate multiple information sources and integrate modular developments. The data warehouse is dedicated to support health care decision-making.


Associations between television watching and car riding behaviors and development of depressive symptoms: a prospective study.
To examine the longitudinal association between sedentary behaviors and risk of development of depressive symptoms. The study population consisted of 4802 participants in the Aerobics Center Longitudinal Study (1012 women and 3790 men) aged 18 to 80 years who did not report depressive moods when they completed a health survey during 1982 in which they reported their time spent watching television (TV) and riding in a car each week. All participants completed a follow-up health survey when they responded to the 10-item Center for Epidemiologic Studies Depression Scale. Those who scored 8 or more on the Center for Epidemiologic Studies Depression Scale were considered to have depressive symptoms. Among the 4802 participants, 568 reported depressive symptoms during a mean follow-up of 9.3 years. After multivariate adjustment including moderate- and vigorous-intensity physical activity, time riding in a car, time watching TV, and combined time spent in the 2 sedentary behaviors were positively associated with depressive symptoms (each P<.05 for trend). Individuals who reported 9 h/wk or more riding in a car, more than 10 h/wk watching TV, or 19 h/wk or more of combined sedentary behavior had 28%, 52%, and 74% greater risk of development of depressive symptoms than those who reported less than 5 h/wk, less than 5 h/wk, or less than 12 h/wk, respectively, after adjusting for baseline covariates and moderate- and vigorous-intensity physical activity. The positive association between time riding in a car or time watching TV and depressive symptoms was only observed among individuals who did not meet the current physical activity guidelines. More time reported in these 2 sedentary behaviors was positively associated with depressive symptoms. However, the direct associations between time spent in car riding and TV viewing and depressive symptoms were only significant among those who did not meet the current physical activity recommendations.


Black abalone (Haliotis cracherodii) population structure shifts through deep time: Management implications for southern California's northern Channel Islands.
For over 10,000 years, black abalone (Haliotis cracherodii) were an important resource in southern California, first for coastal Native Americans, then beginning in the nineteenth century, as one of the state's first commercial shellfisheries. By 1993, after years of heavy fishing, rising sea surface temperatures (SST), and the spread of withering syndrome (WS), black abalone populations declined dramatically, resulting in the closure of the Alta California fishery. After nearly 25 years of management and recovery efforts, black abalone are showing signs of ecological rebound along some Channel Island shorelines. These include the presence of juvenile abalone and increasing densities, largely from data collected by Channel Islands National Park (CINP) monitoring efforts that began in 1985.In an effort to apply deeper historical perspectives to modern fisheries management and restoration, we analyzed black abalone size data from San Miguel Island at prehistoric and historical archeological sites spanning the last 10,000 years and compared these populations to those described by CINP biologists between 1985 and 2013.We found a statistically significant relationship between SST and black abalone size distributions during the ancient record, along with dramatic shifts in population size structure toward larger individuals between the nineteenth century and modern periods. A pattern of larger mean black abalone sizes was identified during warm SSTs, when compared against intervals of cooler SSTs.Synthesis and applications. Our study provides a deep historical perspective of abalone population size distributions, patterns within these distributions through time, and parallels to modern abalone populations. Our results may help managers determine whether the current (and future) size and age structure of intertidal black abalone populations around the northern Channel Islands are "natural" and healthy, measured against the 10,000 year history of black abalone fishing in southern California.


A pilot study: lenticule quality of hyperopic small incision lenticule extraction (SMILE) in rabbits.
To evaluate lenticule surface characteristics of small incision lenticule extraction (SMILE) for hyperopia correction in rabbits. The left and right eyes of 8 rabbits were divided into two groups. The right eyes were assigned to a myopia group, and the left eyes to a hyperopia group. The rabbits received SMILE procedures with + 3.00 D and - 3.00 D correction for the hyperopia and myopia groups, respectively. Extracted lenticules were examined via scanning electron microscopy. Lenticules from odd-numbered rabbits were accessed with the anterior surface, and lenticules from even-numbered rabbits were observed with the posterior surface. A previously established scoring system was used to evaluate lenticule surface characteristics. Statistical analysis was conducted to compare the scores between the two groups. All procedures were performed successfully, and the lenticules were extracted smoothly. One myopia lenticule that was facing downward was handled failed in preparation for imaging, thus 15 lenticules were ultimately graded. Twelve lenticules exhibited smooth surfaces, and regularly arranged tissue bridges were observed in almost all regions. Three lenticules exhibited a partially rough surface and irregularities affecting more than 10% of the lenticules (2 in the hyperopia group and 1 in the myopia group). Rough lenticules occurred in twice as many lenticules in the hyperopia group compared to the myopia group. Scan quality of lenticules after SMILE for hyperopia correction is comparable to that of myopia lenticules. The shape of hyperopic lenticule may increase the difficulty of surgical manipulation and result in surface roughness.


Data Quality and Assessment, Validation Methods and Error Propagation through the Simulation Software: Report from the Round-Table Discussion at the 10th World Congress of Chemical Engineering in Barcelona (October 1-5, 2017).
The issues of data quality and propagation of data uncertainties into process design and plant specifications are of great current interest. Hence, two Working Parties of the European Federation of Chemical Engineers (EFCE) organized a Round Table Discussion on the topic, as part of the World Congress of Chemical Engineering (WCCE10) in Barcelona, in October 2017. The discussion was guided by industrial and academic experts, with the audience as a key part of the discussion, trying to find some answers in three areas: Data acquisition and evaluation of experimental uncertainties, tools for data reconciliation to improve their quality, and impact of data uncertainties on the process at the end. Several concrete stories are presented that demonstrate the importance of considering data quality and all possible contributions to the uncertainty of chemical process design. Difficulties associated with data quality are discussed at various levels: (1) the experimentalists (measurement issues, evaluation of uncertainties, use of consistency analysis tools); (2) model developers (capture of adequate physics, parameter regression strategies, uncertainty propagation), (3) vendors of process simulation software, and (4) process engineers (who are responsible at the end). Paths for improvements were proposed through better and more efficient communication among different participants, as well as through education.


Alpha1-microglobulin as an early biomarker of sepsis-associated acute kidney injury: a prospective cohort study.
Sepsis emerges as the leading risk factor for acute kidney injury (AKI) development in critically ill patients. Much effort has been invested so far on early diagnosis of AKI using promising biomarkers. This study aimed to determine whether urine alpha1-microglobulin (α1m), a lipocaline member previously used as an indicator of proximal tubular dysfunction, can early predict the development of sepsis-associated AKI (SAAKI) in critically ill patients. A prospective, observational study was conducted in a single center Intensive Care Unit (ICU). Patients with normal renal function admitted to the ICU followed for sepsis and AKI development. Urine α1m levels were analyzed in pooled samples from 24-hour urine collections on sepsis onset and at various time points thereafter. The diagnostic performance of urine α1m was assessed using thenonparametriccalculation of the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Among 286 critically ill patients admitted to our ICU in a year, 45 patients with sepsis met the inclusion criteria. SAAKI developed in 16 septic patients (35.6%). Urine α1m levels were significantly elevated in all septic patients (average value of all samples on the day of sepsis: 46.02 ± 7.17 mg/l) and showed a trend to increase in patients who finally developed SAAKI. The AUC for SAAKI prediction according to α1m urine levels 24-hours before SAAKI onset was 0.739 (sensitivity 87.5%, specificity 62.07%, cutoff level 47.9 mg/l). Urine α1m 24-hours before SAAKI, serum creatinine on sepsis onset and Acute Physiology and Chronic Health Evaluation II (APACHE II) score on sepsis onset emerged as the most powerful independent predictors of SAAKI. The combination of these three parameters improved the AUC for SAAKI prediction to 0.944. Urine α1m levels might help in the early prediction of SAAKI development and may prove useful biomarker. The pathogenetic implications of α1m in sepsis and SAAKI need further investigation. Hippokratia 2014; 18 (3): 262-268.


Spatial and temporal trends of contaminants in Canadian Arctic freshwater and terrestrial ecosystems: a review.
The state of knowledge of contaminants in Canadian Arctic biota of the freshwater and terrestrial ecosystems has advanced enormously since the publication of the first major reviews by Lockhart et al. and Thomas et al. in The Science of the Total Environment in 1992. The most significant gains are new knowledge of spatial trends of organochlorines and heavy metal contaminants in terrestrial animals, such as caribou and mink, and in waterfowl, where no information was previously available. Spatial trends in freshwater fish have been broadened, especially in the Yukon, where contaminant measurements of, for example, organochlorines were previously non-existent. A review of contaminants data for fish from the Northwest Territories, Yukon and northern Quebec showed mercury as the one contaminant which consistently exceeds guideline limits for subsistence consumption or commercial sale. Lake trout and northern pike in the Canadian Shield lakes of the Northwest Territories and northern Quebec generally had the most elevated levels. Levels of other heavy metals were generally not elevated in fish. Toxaphene was the major organochlorine contaminant in all fish analyzed. The concentrations of organochlorine contaminants in fish appear to be a function not only of trophic level but of other aspects of the lake ecosystem. Among Arctic terrestrial mammals, PCBs and cadmium were the most prominent contaminants in the species analyzed. Relatively high levels (10-60 micrograms g-1) of cadmium were observed in kidney and liver of caribou from the Yukon, the Northwest Territories and northern Quebec, with concentrations in western herds being higher than in those from the east. For the organochlorine contaminants, a west to east increase in zigma PCBs, HCB and zigma HCH was found in caribou, probably as a result of the predominant west to east/north-east atmospheric circulation pattern which delivers these contaminants from industrialized regions of central and eastern North America to the Arctic via long-range atmospheric transport. Radiocesium contamination of lichens and caribou has continued to decrease. Significant contamination by PCBs and lead of soils and vascular plants was observed in the immediate vicinity and within a 20-km radius of DEW line sites in the Canadian Arctic. There was also evidence for transfer of PCBs from plants to lemmings. There was no evidence, however, that large mammals such as caribou living in the general area of the DEW line sites had elevated levels of PCBs. There is very limited temporal trend information for most contaminants in biota of Arctic terrestrial and freshwater environments.


Tackling differential attainment in specialist GP training in England and Scotland.
In this article NHS England and NHS Education for Scotland describe practical ways we are tackling differences in the attainment of people training as general practitioners (GPs).Trainees from minority ethnic groups and international medical graduates are less likely than others to qualify as GPs. It is difficult to change systemic inequalities, but over the past five years we have made practical changes to GP speciality training. Educators recognise there is an issue and are trying to tackle it.For example, people who had not successfully qualified had an opportunity to return to GP training. When we provided individualised targeted support, the proportion who completed training significantly increased (76%).This was a catalyst for reviewing unconscious bias in GP training. We implemented a national programme to tackle differential attainment and system-level bias. Educators now work with all GP trainees to identify their individual needs. Supervisors are trained to recognise bias and provide targeted support. There is mental health support and regular reviews to see whether trainees are ready to sit exams. Trainee representatives are championing the learner voice in national committees. Exams are being altered to reduce unconscious bias. We are monitoring attainment over time.The key message is that differential attainment should not be in the 'too hard basket'. The narrative is changing from 'can't do' to 'must do', supported by appropriate leadership, promotion and resourcing. There is much more to do, but we are making changes, evaluating and applying our learning. We have moved from talking to taking action.


Selective deletion of GRK2 alters psychostimulant-induced behaviors and dopamine neurotransmission.
GRK2 is a G protein-coupled receptor kinase (GRK) that is broadly expressed and is known to regulate diverse types of receptors. GRK2 null animals exhibit embryonic lethality due to a severe developmental heart defect, which has precluded the study of this kinase in the adult brain. To elucidate the specific role of GRK2 in the brain dopamine (DA) system, we used a conditional gene knockout approach to selectively delete GRK2 in DA D1 receptor (D1R)-, DA D2 receptor (D2R)-, adenosine 2A receptor (A2AR)-, or DA transporter (DAT)-expressing neurons. Here we show that select GRK2-deficient mice display hyperactivity, hyposensitivity, or hypersensitivity to the psychomotor effects of cocaine, altered striatal signaling, and DA release and uptake. Mice with GRK2 deficiency in D2R-expressing neurons also exhibited increased D2 autoreceptor activity. These findings reveal a cell-type-specific role for GRK2 in the regulation of normal motor behavior, sensitivity to psychostimulants, dopamine neurotransmission, and D2 autoreceptor function.


Factors associated with bicycle-helmet use among 8-16 years aged Turkish children: a questionnaire survey.
The purpose of this cross-sectional, descriptive study was to assess the rates of helmet and to examine variables related to bicycle helmet use in a sample of 8-16 year old Turkish children. Data were collected from a purposive sample of 1180 students who self-identified as cyclists and 1128 parents or guardians in two state primary schools in two urban municipalities in the northern part of Istanbul, Turkey. Results showed that self-reported helmet use was found prevalent in 4.4% of the children. The most cited reasons for not wearing a helmet were: 'don't own one', 'helmets are uncool', and 'uncomfortable'. Significant differences between helmet users and non-users correlated to gender, age, owning a bicycle helmet, wearing a friend's bicycle helmet and parents or guardian's bicycle helmet use while riding. Only three variables--helmet ownership (OR = 10.028, 95% CI 5.08, 19.79), parents' helmet use (OR = 2.62, 95% CI 1.22, 5.66) and friends' helmet use (OR = 0.16, 95% CI 0.07, 0.37) emerged as significant predictors of the likelihood of helmet use. The relatively low helmet use prevalence points to an urgent need for a multipronged campaign, including strategies such as raising awareness, educating primarly parents and friends, and distributing bicycle helmets for free or at a reduced cost.


Growth management of vetiver (Vetiveria zizanioides) under Mediterranean conditions.
In spite of the advantages of Vetiver grass in light of environmental aspects, this plant is not used in the Mediterranean region. The objectives of the present study were: (i) to elucidate growth parameters and establishment of Vetiver under Mediterranean conditions suitable for its various environmental applications; and (ii) to develop management practices for growing vetiver under Mediterranean conditions. In greenhouse experiments conducted under controlled conditions it was found that, in general, increasing the minimum/maximum temperatures to 21-29 degrees C significantly increased plant height. In the Mediterranean region, this range of air temperatures is obtained mainly during the summer, from June to September. For air temperatures up to 15-23 degrees C the effect of day length on plant height was insignificant, whereas in air temperature >15-23 degrees C, the plant heights under long day conditions were significantly higher than under short day. The number of sprouts per plant increased exponentially with increasing air temperature, and was not significantly affected by the day length at any air temperature range. In open fields, the heights of irrigated vetiver plants were significantly higher than those of rain-fed plants. It was concluded that, once they were established, vetiver plants could survive the dry summer of the Mediterranean region under rain-fed conditions, but they would be shorter than under irrigation. Cutting or burning of the plant foliage during the spring did not improve the survival of vetiver during the dry summer. In order to obtain fast growth of vetiver and to increase the possibility of its using the rainwater, the plants should be planted in the winter, during February and March. However, under this regime, the vetiver plant cannot be used as a soil stabilizer during the first winter, because the plant is still small. In contrast, under irrigation it is advantageous to plant vetiver at the beginning of the summer; the plant then has sufficient time to grow and develop before the beginning of the winter, so that its effect as a soil stabilizer in the following wet winter could be maximal. It was found that vetiver could grow in a wide range of substrates, such as: sandy soil, loamy sand, clay soil, crushed limestone, sandy clay loam, and tuff/peat mixture.


Systematic Reconstruction of the Complete Two-Component Sensorial Network in Staphylococcus aureus.
In bacteria, adaptation to changes in the environment is mainly controlled through two-component signal transduction systems (TCSs). Most bacteria contain dozens of TCSs, each of them responsible for sensing a different range of signals and controlling the expression of a repertoire of target genes (regulon). Over the years, identification of the regulon controlled by each individual TCS in different bacteria has been a recurrent question. However, limitations associated with the classical approaches used have left our knowledge far from complete. In this report, using a pioneering approach in which a strain devoid of the complete nonessential TCS network was systematically complemented with the constitutively active form of each response regulator, we have reconstituted the regulon of each TCS of S. aureus in the absence of interference between members of the family. Transcriptome sequencing (RNA-Seq) and proteomics allowed us to determine the size, complexity, and insulation of each regulon and to identify the genes regulated exclusively by one or many TCSs. This gain-of-function strategy provides the first description of the complete TCS regulon in a living cell, which we expect will be useful to understand the pathobiology of this important pathogen.IMPORTANCE Bacteria are able to sense environmental conditions and respond accordingly. Their sensorial system relies on pairs of sensory and regulatory proteins, known as two-component systems (TCSs). The majority of bacteria contain dozens of TCSs, each of them responsible for sensing and responding to a different range of signals. Traditionally, the function of each TCS has been determined by analyzing the changes in gene expression caused by the absence of individual TCSs. Here, we used a bacterial strain deprived of the complete TC sensorial system to introduce, one by one, the active form of every TCS. This gain-of-function strategy allowed us to identify the changes in gene expression conferred by each TCS without interference of other members of the family.


Jacalin-carbohydrate interactions: distortion of the ligand molecule as a determinant of affinity.
Jacalin is among the most thoroughly studied lectins. Its carbohydrate-binding site has also been well characterized. It has been postulated that the lower affinity of β-galactosides for jacalin compared with α-galactosides is caused by steric interactions of the substituents in the former with the protein. This issue has been explored energetically and structurally using different appropriate carbohydrate complexes of jacalin. It turns out that the earlier postulation is not correct. The interactions of the substituent with the binding site remain essentially the same irrespective of the anomeric nature of the substitution. This is achieved through a distortion of the sugar ring in β-galactosides. The difference in energy, and therefore in affinity, is caused by a distortion of the sugar ring in β-galactosides. The elucidation of this unprecedented distortion of the ligand as a strategy for modulating affinity is of general interest. The crystal structures also provide a rationale for the relative affinities of the different carbohydrate ligands for jacalin.


Running Economy in the Vertical Kilometer.
New and promising variables are being developed to analyze performance and fatigue in trail running, such as mechanical power, metabolic power, metabolic cost of transport and mechanical efficiency. The aim of this study was to analyze the behavior of these variables during a real vertical kilometer field test. Fifteen trained trail runners, eleven men (from 22 to 38 years old) and four women (from 19 to 35 years old) performed a vertical kilometer with a length of 4.64 km and 835 m positive slope. During the entire race, the runners were equipped with portable gas analyzers (Cosmed K5) to assess their cardiorespiratory and metabolic responses breath by breath. Significant differences were found between top-level runners versus low-level runners in the mean values of the variables of mechanical power, metabolic power and velocity. A repeated-measures ANOVA showed significant differences between the sections, the incline and the interactions between all the analyzed variables, in addition to differences depending on the level of the runner. The variable of mechanical power can be statistically significantly predicted from metabolic power and vertical net metabolic COT. An algebraic expression was obtained to calculate the value of metabolic power. Integrating the variables of mechanical power, vertical velocity and metabolic power into phone apps and smartwatches is a new opportunity to improve performance monitoring in trail running.


Flow cytometric identification of intracellular antigens: detection of minimal residual leukemia.
Recent advances in preparation of cells for flow cytometric analysis have enabled the sensitive detection of intracellular antigens. We have examined the utility of two color flow cytometry for the detection of minimal residual T cell acute lymphoblastic leukemia (T-ALL) using a combination of expression of the pan-T cell marker CD5 and intranuclear terminal deoxynucleotidyl transferase (TdT). CD5+TdT+ cells can be sensitively detected above background in remission bone marrows (0.03% sensitivity). Possible extension of this technique to the detection of minimal residual B-lineage ALL, acute myelogenous leukemia (AML), and non-hematologic malignancies is discussed.


Hidden heterogeneity and circadian-controlled cell fate inferred from single cell lineages.
The origin of lineage correlations among single cells and the extent of heterogeneity in their intermitotic times (IMT) and apoptosis times (AT) remain incompletely understood. Here we developed single cell lineage-tracking experiments and computational algorithms to uncover correlations and heterogeneity in the IMT and AT of a colon cancer cell line before and during cisplatin treatment. These correlations could not be explained using simple protein production/degradation models. Sister cell fates were similar regardless of whether they divided before or after cisplatin administration and did not arise from proximity-related factors, suggesting fate determination early in a cell's lifetime. Based on these findings, we developed a theoretical model explaining how the observed correlation structure can arise from oscillatory mechanisms underlying cell fate control. Our model recapitulated the data only with very specific oscillation periods that fit measured circadian rhythms, thereby suggesting an important role of the circadian clock in controlling cellular fates.


Copper(II) and gallium(III) complexes of trans-bis(2-hydroxybenzyl) cyclen derivatives: absence of a cross-bridge proves surprisingly more favorable.
Two cyclen (1,4,7,10-tetraazacyclododecane) derivatives bearing trans-bis(2-hydroxybenzyl) arms, the 1,7-(2-hydroxybenzyl)-1,4,7,10-tetraazacyclododecane (H2do2ph) and its cross-bridged counterpart (H2cb-do2ph), have been synthesized, aiming toward the possible use of their copper(II) and gallium(III) complexes in nuclear medicine. The protonation of both compounds was studied in aqueous solution as well as their complexes with Cu(2+) and Ga(3+) cations. The complexes of both ligands with Ca(2+) and Zn(2+) metal ions were also studied due to the abundance of these cations in biological media. In mild conditions the complexes of Ca(2+) and Ga(3+) with H2cb-do2ph did not form. The behavior of the two ligands and their complexes was compared by the values of the equilibrium constants, the data of varied spectroscopic techniques, the values of redox potentials of their copper(II) complexes, and the resistance of the complexes to acid dissociation. It was expected that, as found for related pairs of cyclen and cyclam (1,4,8,11-tetraazacyclotetradecane) derivatives, the cross-bridged macrocyclic derivative could be an excellent ligand for the complexation of copper(II). Additionally, the N-2-hydroxybenzyl groups were chosen due to their known ability to coordinate the gallium(III) cation. Due to the small size of the latter cation and its particular propensity to form hexacoordinate complexes, it was also expected that there would be a good ability of both ligands for the uptake of Ga(3+). Surprisingly, the results revealed that the cyclen derivative H2do2ph is the best ligand for the coordination of Cu(2+) and Ga(3+) cations, not only from their thermodynamic stability as expected but also from their kinetic inertness, when compared with its cross-bridged counterpart.


Haemophagocytic lymphohistiocytosis occurred during induction chemotherapy in an acute monocytic leukemia patient with FLT3-ITD and DNMT3A mutations.
Haemophagocytic lymphohistiocytosis (HLH) is considered to be a large challenge for clinicians due to the variable overlaps of symptoms with other severe diseases and a high rate of mortality. Prompt diagnosis and treatment are crucial to avoid a fatal outcome. However, very limited reports have focused on HLH during chemotherapy (Ch-HLH) due to a low incidence and insufficient knowledge. A 22-year-old male was diagnosed with acute monocytic leukemia with FLT3-ITD and DNMT3A mutations and pulmonary infection. He received IA regimen (Idarubicin, 8 mg/m2/d for 3 days and cytarabine, 100 mg/m2/d for 7 days) chemotherapy, anti-infection drugs and blood components transfusions. During the stage of bone marrow suppression, he presented with a fever, cytopenia (WBC, 0.43 × 109/L; Hb, 73 g/L and PLT, 1 × 109/L), refractory coagulation dysfunction (APTT, 104.0 s; PT, 30.5 s and Fbg, 0.87 g/L), splenomegaly (3 cm below the costal margin), hyperferritinemia (SF > 3000 μg/L), increased soluble interleukin-II receptors (sIL-2R > 7500 u/mL) and haemophagocytosis in the bone marrow and was diagnosed with HLH. After he was treated with methylprednisolone at 500 mg/d for 3 days, 120 mg/d for 3 days and 80 mg/d for 3 days, followed by a gradually reduced dose combined with powerful anti-infection drugs, his symptoms subsided and his abnormal parameters recovered to normal levels. Patients with HLH in acute leukemia have a high rate of mortality. This case report provides helpful clinical experiences relative to the recognition and treatment of Ch-HLH for clinicians.


Relationships between spinal landmarks and skin surface markers.
This investigation determined relationships between coronal vertical alignment (CVA) and sagittal vertical alignment (SVA) variables calculated from radiographs and surface markers representing bony landmarks. Biplanar radiographs were taken on 28 subjects (standing) after 2 metallic surface markers were placed on the skin superficial to C7 and S2. The CVA-R and SVA-R were measured on the radiographs. Similar variables were calculated from the surface markers (CVA-P-R, SVA-P-R). Correlation between CVA-R and CVA-P-R was 0.894 (p<0.000), and between SVA-R and SVA-P-R was 0.946 (p<0.000). Results lead to three recommendations: (1) obtain surface marker data when radiographs are taken to establish relationships between the two sets of data, (2) take care in providing instructions to the subjects if measures are to be taken at different times, and (3) observe caution in interpreting results when simultaneous x-ray and surface marker data were not recorded.


Quantifying clinical change: discrepancies between patients' and providers' perspectives.
Interpreting the clinical significance of changes in patient-reported outcomes (PROs) is critically important. The most commonly used approach is to anchor mean changes on PRO scores against a global assessment of change. Whether the assessor of global change should be patients or their physicians is unknown. We compared patients' and physicians' assessments of change over time to examine which was more aligned with patients' changes in PRO measures. A total of 459 chronic heart failure patients aged >30 years were enrolled from 13 US centers. Data were obtained by medical record abstraction, physical assessments, and patient interviews at a baseline clinic visit and 6 weeks later. Health status was measured with the disease-specific Kansas City Cardiomyopathy Questionnaire (KCCQ), and both patients and physicians completed a validated 15-level global assessment of change, ranging from large deterioration to large improvement. There was substantial variation between physicians/patients' global assessment of clinical change (weighted kappa = 0.36, 95 % CI 0.28, 0.43). Overall, physician assessments were more strongly correlated with change on the KCCQ summary score than were patients' assessments (physician R = 0.37, patient R = 0.29). There was substantial variation between patients' and physicians' global assessment of 6-week change in heart failure status. Physician assessments of the importance of clinical changes were more strongly associated with changes in all domains of patient-reported health status, as assessed by the KCCQ, and may provide a more consistent method for defining the clinical importance of changes in patients' health status.


Regulation of the formation of tumor cell pseudopodia by the Na(+)/H(+) exchanger NHE1.
The Na(+)/H(+) exchanger NHE1 is involved in intracellular pH homeostasis and cell volume regulation and accumulates with actin in the lamellipodia of fibroblasts. In order to determine the role of NHE1 following epithelial transformation and the acquisition of motile and invasive properties, we studied NHE1 expression in polarized MDCK cells, Moloney Sarcoma virus (MSV) transformed MDCK cells and an invasive MSV-MDCK cell variant (MSV-MDCK-INV). Expression of NHE1 was significantly increased in MSV-MDCK-INV cells relative to MSV-MDCK and MDCK cells. NHE1 was localized with b-actin to the tips of MSV-MDCK-INV cell pseudopodia by immunofluorescence. Sensitivity of NHE1-mediated (22)Na uptake to ethylisopropylamiloride, a specific inhibitor of NHE1, was increased in MSV-MDCK cells relative to MDCK cells. Changes in intracellular pH induced upon EIPA treatment were also of higher magnitude in MSV-MDCK and MSV-MDCK-INV cells compared to wild-type MDCK cells, especially in Hepes-buffered DMEM medium. Inhibition of NHE1 by 50 microM ethylisopropylamiloride induced the disassembly of actin stress fibers and redistribution of the actin cytoskeleton in all cell types. However, in MSV-MDCK-INV cells, the effect of ethylisopropylamiloride treatment was more pronounced and associated with the increased reversible detachment of the cells from the substrate. Videomicroscopy of MSV-MDCK-INV cells revealed that within 20 minutes of addition, ethylisopropylamiloride induced pseudopodial retraction and inhibited cell motility. The ability of ethylisopropylamiloride to prevent nocodazole-induced formation of actin stress fibers in MSV-MDCK cells was more pronounced in Hepes medium relative to NaHCO(3) medium, showing that NHE1 can regulate actin stress fiber assembly in transformed MSV-MDCK cells via its intracellular pH regulatory effect. These results implicate NHE1 in the regulation of the actin cytoskeleton dynamics necessary for the adhesion and pseudopodial protrusion of motile, invasive tumor cells.


Relative abundance of two molecular forms of Na+,K+-ATPase in the ferret heart: developmental changes and associated alterations of digitalis sensitivity.
We have previously reported that adult ferret hearts have high affinity and low affinity ouabain-binding sites in approximately equal abundance corresponding, respectively, to alpha(+) and alpha subunits of Na+,K+-ATPase. In the present study, possible changes in relative abundance of the two molecular forms of Na+,K+-ATPase during postnatal development were examined. In 5-day-old or 7- to 10-day-old ferret heart, there was a predominance of the alpha form of Na+,K+-ATPase, the alpha(+)/alpha ratio being approximately 1:7 and 1:4 at 5 and 7-10 days, respectively, in contrast to a 1:1 ratio observed previously in adult heart. Corresponding to the reduced number of high affinity ouabain-binding sites associated with the alpha(+) form of Na+,K+-ATPase, ouabain sensitivity of the Na+,K+-ATPase isolated from newborn ferret heart was lower than that of adult ferret heart. Furthermore, the low dose positive inotropic effect, which was apparent in the adult ferret heart, could not be observed in the newborn ferret heart. The present finding may represent a plausible explanation for low digitalis sensitivity of neonatal heart observed in several mammalian species.


Factors in rehospitalisation for severe pressure ulcer care in spinal cord injury/disorders.
Repeated hospital admissions (RHA) for ongoing pressure ulcer (PU) care remains a significant challenge in the clinical management of the spinal cord injury/disorders (SCI/D) population. The current study investigated the significance of risk factors for PU treatment and RHA. A retrospective chart review of veterans admitted to the Louis Stokes Cleveland Department of Veterans Affairs Medical Center (LSCDVAMC) Spinal Cord Injury (SCI) unit for Category III or IV PUs was carried out. A random sample of 105 individuals with SCI/D, evaluated by the Wound Care Team (WCT) from 2006 to 2009 was assessed. Multiple PU development risk factors were extracted from the electronic health record system using standardised data collection forms and entered into the Spinal Cord Injury Pressure Ulcer Database (SCIPUD). Potential associations with RHA were analysed. Twenty variables were initially identified as potentially related to PU development. Descriptive statistics and statistically significant associations between risk factors and RHA were determined. Demographic factors showed no significant association with RHA. Duration of injury, power wheelchair use and sub-optimally managed spasticity (SMS) were significantly associated with higher RHA. Sub-optimally managed neurogenic bowel (SMNB) at admission was significantly associated with reduced RHA. Factors previously found to be predictive of initial PU development may not, in fact, be predictive of RHA. Some protective trends were observed, such as polypharmacy and marital status, but these did not reach statistical significance in this preliminary study of admission characteristics, warranting further research. There were no external sources of funding for this study. The authors have no conflicts of interests to declare.


Photoinduced electron transfer between cytochrome c peroxidase and horse cytochrome c labeled at specific lysines with (dicarboxybipyridine)(bisbipyridine)ruthenium(II)
The reactions of yeast cytochrome c peroxidase with horse cytochrome c derivatives labeled at specific lysine amino groups with (dicarboxybipyridine)(bisbipyridine)ruthenium(II) [Ru(II)] were studied by flash photolysis. All of the derivatives formed complexes with cytochrome c peroxidase compound I (CMPI) at low ionic strength (2 mM sodium phosphate, pH 7). Excitation of Ru(II) to Ru(II*) with a short laser flash resulted in electron transfer to the ferric heme group in cytochrome c, followed by electron transfer to the radical site in CMPI. This reaction was biphasic and the rate constants were independent of CMPI concentration, indicating that both phases represented intracomplex electron transfer from the cytochrome c heme to the radical site in CMPI. The rate constants of the fast phase were 5200, 19,000, 55,000, and 14,300 s-1 for the derivatives modified at lysines 13, 25, 27, and 72, respectively. The rate constants of the slow phase were 260, 520, 200, and 350 s-1 for the same derivatives. These results suggest that there are two binding orientations for cytochrome c on CMPI. The binding orientation responsible for the fast phase involves a geometry that supports rapid electron transfer, while that for the slow phase allows only slow electron transfer. Increasing the ionic strength up to 40 mM increased the rate constant of the slow phase and decreased that of the fast phase. A single intracomplex electron transfer phase with a rate constant of 2800 s-1 was observed for the lysine 72 derivative at this ionic strength. When a series of light flashes was used to titrate CMPI to CMPII, the reaction between the cytochrome c derivative and the Fe(IV) site in CMPII was observed. The rate constants for this reaction were 110, 250, 350, and 140 s-1 for the above derivatives measured in low ionic strength buffer.


Cortical area V4 is critical for certain texture discriminations, but this effect is not dependent on attention.
This study examined the question of which features of a complex grouping discrimination make it vulnerable to permanent elimination by V4 lesions. We first verified that the line element grouping discrimination, which we previously reported to be devastated by V4 lesions, was similarly affected in the monkeys of this study. The permanence of the deficit was established by mapping its visual field distribution and then testing this discrimination for an extended period at a locus on the border of the deficit. Also, a staircase procedure was used to provide the monkey with within session instruction in the grouping discrimination, but this did not improve V4 lesion performance. Grouping was then compared with several discriminations that shared some features with it, but which were found not to be permanently eliminated by V4 lesions. This comparison suggested that grouping (rather than segmentation or response to a single element) was one feature that made the discrimination vulnerable, a second was the similarity in shape of the texture elements to be grouped. Finally, we tested visual crowding, a property of peripheral vision that is thought to reflect neuronal interactions early in visual cortex, possibly in area V1, and found no effect of V4 lesions. A control experiment with human observers tested whether the elimination of grouping by V4 lesions might be due to an alteration of attention, but found no evidence to support this hypothesis. These results show that severe disruption of texture discriminations by V4 lesions depends on both the nature of the discrimination and the type of texture elements involved, but does not necessarily involve the disruption of attention.


Cathepsin D Attenuates the Proliferation of Vascular Smooth Muscle Cells Induced by the AGE/RAGE Pathway by Suppressing the ERK Signal.
In this study, we aimed to clarify the role and mechanism by which Cathepsin D (CTSD) mediates the advanced glycation end products (AGEs)-induced proliferation of vascular smooth muscle cells (VSMCs). We conducted a Western blotting assay and co-immunoprecipitation assay to detect the expression of target proteins and the interaction between different proteins. Cell Counting Kit-8 (CCK-8) assay and 5- ethynyl-2'-deoxyuridine (EdU) were used to evaluate the proliferation. AGEs significantly promoted phenotypic switching and proliferation of VSMCs in a concentration-dependent manner. This effect of AGEs was accompanied by inhibition of CTSD. Both the proliferation of VSMCs and inhibition of CTSD induced by AGEs could be attenuated by the specific inhibitor of the receptor for advanced glycation end products (RAGE), FPS-ZM1. Overexpression of CTSD significantly alleviated these effects of AGEs on VSMCs. The mechanism of CTSD action in VSMCs was also explored. Overexpression of CTSD reduced the activation of p-ERK caused by AGEs. By contrast, the knockdown of CTSD, elicited using a plasmid containing short hairpin RNA (shRNA) against CTSD, further increased the activation of p-ERK compared to AGEs alone. Additionally, co-immunoprecipitation studies revealed an endogenous interaction between CTSD, a protease, and p-ERK, its potential substrate. It has been demonstrated that CTSD downregulates the level of phosphorylated ERK by degrading its target, and this interaction plays a critical role in the proliferation of VSMCs induced by the AGE/RAGE axis. These results provide a novel insight into the prevention and treatment of vascular complications in diabetes.


A pilot study examining the relationship among Crohn disease activity, glucagon-like peptide-2 signalling and intestinal function in pediatric patients.
BACKGROUND⁄ The relationship between the enteroendocrine hormone glucagon-like peptide 2 (GLP-2) and intestinal inflammation is unclear. GLP-2 promotes mucosal growth, decreases permeability and reduces inflammation in the intestine; physiological stimulation of GLP-2 release is triggered by nutrient contact. The authors hypothesized that ileal Crohn disease (CD) affects GLP-2 release. With ethics board approval, pediatric patients hospitalized with CD were studied; controls were recruited from local schools. Inclusion criteria were endoscopy-confirmed CD (primarily of the small intestine) with a disease activity index >150. Fasting and postprandial GLP-2 levels and quantitative urinary recovery of orally administered 3-O-methyl-glucose (active transport) and lactulose⁄mannitol (passive) were quantified during the acute and remission phases. Seven patients (mean [± SD] age 15.3 ± 1.3 years) and 10 controls (10.3 ± 1.6 years) were studied. In patients with active disease, fasting levels of GLP-2 remained stable but postprandial levels were reduced. Patients with active disease exhibited reduced glucose absorption and increased lactulose⁄mannitol recovery; all normalized with disease remission. The change in the lactulose⁄mannitol ratio was due to both reduced lactulose and increased mannitol absorption. These findings suggest that pediatric patients with acute ileal CD have decreased postprandial GLP-2 release, reduced glucose absorption and increased intestinal permeability. Healing of CD resulted in normalization of postprandial GLP-2 release and mucosal functioning (nutrient absorption and permeability), the latter due to an increase in mucosal surface area. These findings have implications for the use of GLP-2 and feeding strategies as a therapy in CD patients; further studies of the effects of inflammation and the GLP-2 axis are recommended.


Influence Of Hulling And Storage Conditions On Maintaining Coffee Quality.
Storage is important in the coffee post-harvest. Determining the maximum period that coffee can remain storaged is important aiming to reduce losses in quality and, consequently, allow the producer to achieve maximum profitability. The aim was to determine the suitable storage period for natural and fully washed coffees, under different conditions. Beans were dried to 11% moisture content after dry processing (natural coffee, dry cherry coffee) and wet processing (parchment coffee, fully washed). Before storage, part of the coffee was hulled and part was not. The coffee was stored under refrigerated air (10ºC and 50% relative humidity) or in an environment at 25ºC. In the periods of 0, 3, 6, and 12 months, samples were taken for sensory, electrical conductivity and tetrazolium evaluation. Refrigerated environment favors conservation of sensory and physiological quality of the natural hulled coffee beans and fully washed coffee. Hulled beans of natural and fully washed coffee stored under refrigerated conditions have the initial quality conserved for up to 12 months and in non-controlled environmental, for up to 3 months. Mechanical damage caused by hulling, associated with the lack of tissue fruit parts, contributes to reduction hulled coffee quality in storage, regardless of the processing.


[Determination of seven strobilurin fungicide residues in Chinese herbs by liquid chromatography-tandem mass spectrometry coupled with solid phase extraction].
An LC-MS/MS method was developed for the simultaneously determination of seven strobilurin fungicide residues in Chinese herbs. The strobilurin fungicides include Z-metominostrobin, kresoxim-methyl, dimoxystrobin, picoxystrobin, pyraclostrobin, azoxystrobin and trifloxystrobin. The sample was extracted with ethyl acetate and cleaned-up by an amino SPE column. The seven strobilurin fungicide residues were separated on a C18 column with gradient elution of 1.0 per thousand formic acid and methanol as mobile phases, and detected by ESI-MS in positive ion and selective reaction monitoring (SRM) mode. External standard method was used to the quantification with good linear relationships (r > or = 0. 996). The LOQs were 2 micro g/kg for dimoxystrobin, picoxystrobin and trifloxystrobin, 4 mciro g/kg for pyraclostrobin and azoxystrobin, 10 micro g/kg for Z-metominostrobin and kresoxim-methyl. The recoveries were from 60.4% to 110% with the RSDs between 1.2% and 17%. The developed method is suitable for the determination and confirmation of the seven strobilurin fungicide residues in the three of Eight Zhes ( Ophiopogon japonicus (Thunb.), Scrophularia ningpoensis Hemsl. and Corydalis yanhusuo W T Wang).


Dissociation of Ca2+ accumulation from protein release in calcium paradox: effect of barium.
Experiments were undertaken to establish whether Ba2+ can substitute for Ca2+ in maintaining the structural and functional integrity of the glycocalyx. Adult rat hearts were perfused with Ca2+-free buffer at 37 degrees C, with and without added Ba2+. Ten minutes later Ba2+-free, Ca-containing medium was reintroduced. Hearts that had been perfused with Ca2+-free medium exhibited a distorted glycocalyx. On reperfusion with Ca2+-containing medium these hearts released protein (P less than 0.001), gained Ca2+ (P less than 0.001), and depleted their tissue stores of ATP (P less than 0.001) and CP (P less than 0.001). Hearts that had been perfused with Ca2+-free, Ba2+-containing medium retained an apparently intact glycocalyx. On reperfusion with Ca2+-containing. Ba2+-free medium they gained Ca2+ but did not lose protein. These results suggest that Ba2+ only partially replaces Ca2+ in maintaining the integrity of the cell surface. It is also concluded that the absence of protein release does not necessarily exclude the occurrence of severe changes in ionic permeability after cell injury.


Loss of p120 catenin and links to mitotic alterations, inflammation, and skin cancer.
Tumor formation involves epigenetic modifications and microenvironmental changes as well as cumulative genetic alterations encompassing somatic mutations, loss of heterozygosity, and aneuploidy. Here, we show that conditional targeting of p120 catenin in mice leads to progressive development of skin neoplasias associated with intrinsic NF-kappaB activation. We find that, similarly, squamous cell carcinomas in humans display altered p120 and activated NF-kappaB. We show that epidermal hyperproliferation arising from p120 loss can be abrogated by IkappaB kinase 2 inhibitors. Although this underscores the importance of this pathway, the role of NF-kappaB in hyperproliferation appears rooted in its impact on epidermal microenvironment because as p120-null keratinocytes display a growth-arrested phenotype in culture. We trace this to a mitotic defect, resulting in unstable, binucleated cells in vitro and in vivo. We show that the abnormal mitoses can be ameliorated by inhibiting RhoA, the activity of which is abnormally high. Conversely, we can elicit such mitotic defects in control keratinocytes by elevating RhoA activity. The ability of p120 deficiency to elicit mitotic alterations and chronic inflammatory responses, that together may facilitate the development of genetic instability in vivo, provides insights into why it figures so prominently in skin cancer progression.


Impact of educational policies on access to health care in Brazil: A cross-sectional study.
Various initiatives to improve access to health care have been implemented internationally. In Brazil, this has included policies intended to expand access to higher education for underrepresented socio-economic groups. These measures have reduced inequalities in access to medical education, but it is not known whether they influence career choices. We examine the effect of these educational policies on physician practice patterns in primary care and/or in medically underserved areas. This is a cross-sectional study that analysed the association between affirmative educational policies and the career choices of physicians who graduated between 2010 and 2015. Multivariate binary regression analysis was used to evaluate the impact of affirmative educational policies on physician career choices, including primary care practice, practice in cities with ≤20 000 inhabitants, and practice in less developed municipalities 4 years after graduation. We identified the practice patterns of 65 304 (82.8%) physicians 4 years after their graduation. Most physicians included in our analysis were female (54.5%), aged ≤27 years (72.4%), born in developed cities and studied in private medical schools. Physicians admitted to medical school based on racial or social access policies were more likely to practice in municipalities with fewer than 20 000 inhabitants (odds ratio [OR]: 1.64; 1.36-1.98) and in primary care (OR: 1.55; 1.35-1.53). Physicians who benefited from scholarships for under-represented socio-economic groups were more likely to practice in small cities (OR: 1.24; 1.07-1.43) and primary care (OR: 1.23; 1.11-1.37). The provision of financial aid also improved the likelihood of practice in primary care and underserved areas. Graduation from medical schools located in smaller cities was associated with practice in municipalities ≤20 000 inhabitants and primary care. Our results demonstrated that educational policies implemented in the Brazilian context are effective in reducing inequities in physician distribution and led to an increase in the number of physicians practicing in primary care.