Optical biopsy with optical coherence tomography: feasibility for surgical diagnostics.
Optical coherence tomography (OCT) is a recently developed compact technology which uses infrared light to perform cross-sectional imaging on a micrometer scale. Since OCT provides imaging at a resolution comparable to conventional histology and does not require direct contact with the tissue surface, a role in real-time surgical diagnostics represents a logical extension. In this work, we test the feasibility of OCT for surgical diagnostics by demonstrating imaging in tissue relevent to microsurgical intervention, a previously undescribed observation. Over 50 sites on nervous, reproductive, and microvascular specimens from 10 patients were examined postmortem with OCT. After imaging, tissue was registered with microinjections of dye, under visible light laser guidance, followed by routine histologic processing to confirm the identity of microstructure. The 16 +/- 1 microm resolution allowed subsurface microstructure to be identified at unprecedented resolution. Structures identified included fascicles of peripheral nerves, the internal elastic membrane of microvessels, and the granular layer of the cerebellum. The ability of OCT to provide micrometer-scale definition of tissue microstructure suggests a role in surgical diagnostics. Future in vivo investigations are merited to establish its utility for morbidity reduction associated with surgical intervention.


Biochemical characterization of arylsulfatase-C isozymes in human fibroblasts.
Arylsulfatase-C and sterol sulfatase were thought to be identical enzymes whose X-linked locus escapes inactivation. However, recent evidence shows that they are not identical but that arylsulfatase-C in human fibroblasts exists in two isozymic forms, designated as slow and fast. We now report that the two forms are enzymatically different. When assayed with an artificial fluorogenic substrate, the slow form showed a pH optimum of 8.00 and a Km of 228 microM. In contrast, the fast form showed a pH optimum of 7.67 and a Km of 86.7 microM with substrate inhibition occurring above 0.33 mM. The heat stability of the fast form was slightly below that of the slow form. Polyclonal antibodies raised against the slow form did not cross-react with the fast form. Hence, the two isozymic forms of arylsulfatase-C are enzymatically and structurally different and the slow form is associated with sterol sulfatase activity.


Assessment of impulse duration thresholds for electrical stimulation of muscles (chronaxy) in dogs.
To determine the electrical impulse duration thresholds (chronaxy) for maximal motor contraction of various muscles without stimulation of pain fibers in dogs. 10 healthy adult Beagles. The dogs were used to assess the minimal intensity (rheobase) required to elicit motor contraction of 11 muscles (5 in the forelimb [supraspinatus, infraspinatus, deltoideus, lateral head of the triceps brachii, and extensor carpi radialis], 5 in the hind limb [gluteus medius, biceps femoris, semitendinosus, vastus lateralis, and tibialis cranialis], and the erector spinae). The rheobase was used to determine the chronaxy for each of the 11 muscles in the 10 dogs; chronaxy values were compared with those reported for the corresponding muscles in humans. Compared with values in humans, chronaxy values for stimulation of AA motor fibers in the biceps femoris and semitendinosus muscles and muscles of the more distal portions of limbs were lower in dogs. For the other muscles evaluated, chronaxy values did not differ between dogs and humans. Application of the dog-specific chronaxy values when performing electrical stimulation for strengthening muscles or providing pain relief is likely to minimize the pain perceived during treatment in dogs.


Charting Hydrogen Bond Anisotropy.
A hydrogen bond (HB) is an essential interaction in countless phenomena, regulating the chemistry of life. HBs are characterized by two features, strength and directionality, with a high degree of heterogeneity across different chemical groups. These characteristics are dependent on the electronic configuration of the atoms involved in the interaction, which, in turn, is influenced strongly by the local molecular environment. Studies based on the analysis of HB in the solid phase, such as X-ray crystallography, suffer from significant biases due to packing forces. These will tend to better describe strong HBs at the expenses of weak ones, which will be either distorted or under-represented. Using quantum mechanics (QM), we calculated interaction energies for about a hundred acceptors and donors in a rigorously defined set of geometries. We performed 180,000 independent QM calculations, covering all relevant angular components, mapping strength and directionality in a context free from external biases, with both single-site and cooperative HBs. By quantifying directionality, we show that there is no correlation with strength; therefore, these two components need to be addressed separately. Results demonstrate that there are very strong HB acceptors (e.g., dimethyl sulfoxide) with nearly isotropic interactions and weak ones (e.g., thioacetone) with a sharp directional profile. Similarly, groups can have comparable directional propensity but be very distant in the strength spectrum (e.g., thioacetone and pyridine). Results provide a new perspective on the way HB directionality is described, with implications for biophysics and molecular recognition that ultimately can influence chemical biology, protein engineering, and drug design.


Analysis of adrenoceptor blockade and hypotension elicited by urapidil and prazosin in conscious rat.
The adrenoceptor blocking properties and hypotensive effects of the alpha-1 blocking agents urapidil and prazosin were compared in conscious instrumented rats. Both urapidil and prazosin in i.v. doses of 3 and 6 mg/kg and 0.125 and 0.25 mg/kg, respectively, blocked the pressor response to the alpha-1 adrenoceptor agonist phenylephrine. The hypotensive effects of urapidil and prazosin in these doses were equivalent. Mechanisms of the blood pressure compensation to urapidil-induced hypotension were examined with the arginine vasopressin (AVP) antagonist, d(CH2)5Tyr(Me)AVP, and the angiotensin converting enzyme inhibitor, MK-422. Urapidil alone (6 mg/kg i.v.) and after the AVP antagonist caused a similar degree of hypotension (22 and 15% decrease, respectively, in mean blood pressure). When the alpha-1 antagonist was administered after converting enzyme inhibition, it caused a significantly greater decrease in blood pressure (37%) than in the previous two groups, but not different from that obtained after combined AVP antagonism and converting enzyme inhibition (30% decrease). These results indicate that the renin-angiotensin system compensates for the hypotensive effect due to alpha-1 adrenoceptor blockade, whereas AVP, even assuming that the circulating level was increased by urapidil, was without effect on blood pressure.


Evolutionary insights into the high worldwide prevalence of MBL2 deficiency alleles.
Human mannose-binding lectin (MBL) is a member of the collectin protein family that binds a broad range of microorganisms and activates the lectin-complement pathway of innate immunity. Common alleles of MBL2 disrupt the MBL protein or modulate the amount of protein produced, resulting in MBL deficiency. The clinical manifestations of MBL deficiency have been extensively studied but the actual role of this lectin in immunity to infection remains a matter of strong debate. MBL is commonly thought to play a key role in protective immunity, because MBL deficiency has been associated with an increase in susceptibility to infectious diseases. However, the high worldwide prevalence of multiple MBL2 deficiency or low-producing alleles suggests the converse that MBL deficiency confers protection. To explore the underlying forces accounting for the high worldwide prevalence of MBL2 deficiency alleles, we characterized genetic diversity in and around the MBL2 genomic region in 1166 chromosomes from 24 worldwide populations. Our results clearly demonstrate that the patterns of MBL2 variation are compatible with neutral evolution, as opposed to negative, positive or balanced natural selection. The high worldwide frequencies of MBL2 alleles associated with the production of little or no protein therefore result exclusively from human migration and genetic drift. The evolutionary neutrality of MBL2 strongly supports the notion that MBL2 variation does not have strong effects on population fitness, suggesting, therefore, that this lectin is largely redundant in host human defences.


Development and usability of educational material about workplace particulate matter exposure.
Particulate matter (PM) exposure is an important health risk, both in daily life and in the workplace. It causes respiratory and cardiovascular diseases and results in 800,000 premature deaths per year worldwide. In earlier research, we assessed workers' information needs regarding workplace PM exposure, the properties and effects of PM, and the rationale behind various means of protection. We also concluded that workers do not always receive appropriate risk communication tools with regards to PM, and that their PM knowledge appears to be fragmented and incomplete. We considered several concepts for use as an educational material based on evaluation criteria: ease of use, costs, appropriateness for target audiences and goals, interactivity, implementation issues, novelty, and speed. We decided to develop an educational folder, which can be used to inform employees about the properties, effects and prevention methods concerning PM. Furthermore, we decided on a test setup of a more interactive way of visualisation of exposure to PM by means of exposimeters. For the development of the folder, we based the information needs on our earlier mental models-based research. We adjusted the folder based on the results of ten semi-structured interviews evaluating its usability. The semi-structured interviews yielded commentaries and suggestions for further improvement, which resulted in a number of alterations to the folder. However, in most cases the folder was deemed satisfactory. Based on this study, the folder we developed is suitable for a larger-scale experiment and a practical test. Further research is needed to investigate the efficacy of the folder and the application of the exposimeter in a PM risk communication system.


Insurance cancellations in context: stability of coverage in the nongroup market prior to health reform.
Recent cancellations of nongroup health insurance plans generated much policy debate and raised concerns that the Affordable Care Act (ACA) may increase the number of uninsured Americans in the short term. This article provides evidence on the stability of nongroup coverage using US census data for the period 2008-11, before ACA provisions took effect. The principal findings are threefold. First, this market was characterized by high turnover: Only 42 percent of people with nongroup coverage at the outset of the study period retained that coverage after twelve months. Second, 80 percent of people experiencing coverage changes acquired other insurance within a year, most commonly from an employer. Third, turnover varied across groups, with stable coverage more common for whites and self-employed people than for other groups. Turnover was particularly high among adults ages 19-35, with only 21 percent of young adults retaining continuous nongroup coverage for two years. Given estimates from 2012 that 10.8 million people were covered in this market, these results suggest that 6.2 million people leave nongroup coverage annually. This suggests that the nongroup market was characterized by frequent disruptions in coverage before the ACA and that the effects of the recent cancellations are not necessarily out of the norm. These results can serve as a useful pre-ACA baseline with which to evaluate the law's long-term impact on the stability of nongroup coverage.


Computed Tomography Perfusion-Based Machine Learning Model Better Predicts Follow-Up Infarction in Patients With Acute Ischemic Stroke.
Prediction of infarct extent among patients with acute ischemic stroke using computed tomography perfusion is defined by predefined discrete computed tomography perfusion thresholds. Our objective is to develop a threshold-free computed tomography perfusion-based machine learning (ML) model to predict follow-up infarct in patients with acute ischemic stroke. Sixty-eight patients from the PRoveIT study (Measuring Collaterals With Multi-Phase CT Angiography in Patients With Ischemic Stroke) were used to derive a ML model using random forest to predict follow-up infarction voxel by voxel, and 137 patients from the HERMES study (Highly Effective Reperfusion Evaluated in Multiple Endovascular Stroke Trials) were used to test the derived ML model. Average map, Tmax, cerebral blood flow, cerebral blood volume, and time variables including stroke onset-to-imaging and imaging-to-reperfusion time, were used as features to train the ML model. Spatial and volumetric agreement between the ML model predicted follow-up infarct and actual follow-up infarct were assessed. Relative cerebral blood flow <0.3 threshold using RAPID software and time-dependent Tmax thresholds were compared with the ML model. In the test cohort (137 patients), median follow-up infarct volume predicted by the ML model was 30.9 mL (interquartile range, 16.4-54.3 mL), compared with a median 29.6 mL (interquartile range, 11.1-70.9 mL) of actual follow-up infarct volume. The Pearson correlation coefficient between 2 measurements was 0.80 (95% CI, 0.74-0.86, P<0.001) while the volumetric difference was -3.2 mL (interquartile range, -16.7 to 6.1 mL). Volumetric difference with the ML model was smaller versus the relative cerebral blood flow <0.3 threshold and the time-dependent Tmax threshold (P<0.001). A ML using computed tomography perfusion data and time estimates follow-up infarction in patients with acute ischemic stroke better than current methods.


Radioimmunoassay for the measurement of thrombospondin in plasma and breast cyst fluid: validation and clinical application.
Thrombospondin is an adhesive protein that has been implicated in malignancy, specifically in tumour progression and angiogenesis. We developed a radioimmunoassay for the measurement of thrombospondin in plasma and breast cyst fluid. The assay exhibited high accuracy, with recoveries of 102-136% and acceptable imprecision, with an intra-assay coefficient of variation (CV) of <7.5% across the analytical range 30-1000 ng/mL and inter-assay CV of 4.4% and 7.7% at 152 and 224 ng/mL, respectively. Thrombospondin measured in the breast cyst fluid of patients with gross cystic disease of the breast showed that patients with type II (Na+) cysts had significantly higher concentrations than type I (K+) cysts. The plasma thrombospondin reference range was determined as 131-274 ng/mL. Patients with breast cancer had significantly higher plasma thrombospondin concentrations than normal individuals or patients with benign breast disease. Plasma thrombospondin was higher in breast cancer patients with lymph node involvement.


Development and validation of a life expectancy estimator for multimorbid older adults: a cohort study protocol.
Older multimorbid adults have a high risk of mortality and a short life expectancy (LE). Providing high-value care and avoiding care overuse, including of preventive care, is a serious challenge among multimorbid patients. While guidelines recommend to tailor preventive care according to the estimated LE, there is no tool to estimate LE in this specific population. Our objective is therefore to develop an LE estimator for older multimorbid adults by transforming a mortality prognostic index, which will be developed and internally validated in a prospective cohort. We will analyse data of the Optimising Therapy to Prevent Avoidable Hospital Admissions in Multimorbid Older People cohort study in Bern, Switzerland. 822 participants were included at hospitalisation with age of 70 years or older, multimorbidity (three or more chronic medical conditions) and polypharmacy (use of five drugs or more for >30 days). All-cause mortality will be assessed during 3 years of follow-up. We will apply a flexible parametric survival model with backward stepwise selection to identify the mortality risk predictors. The model will be internally validated using bootstrapping techniques. We will derive a point-based risk score from the regression coefficients. We will transform the 3-year mortality prognostic index into an LE estimator using the Gompertz survival function. We will perform a qualitative assessment of the clinical usability of the LE estimator and its application. We will conduct the development and validation of the mortality prognostic index following the Prognosis Research Strategy (PROGRESS) framework and report it following the Transparent Reporting of a Multivariable Prediction Model for Individual Prognosis or Diagnosis (TRIPOD) statement. Written informed consent by patients themselves or, in the case of cognitive impairment, by a legal representative, was required before enrolment. The local ethics committee (Kantonale Ethikkommission Bern) has approved the study. We plan to publish the results in peer-reviewed journals and present them at national and international conferences.


[Atmospheric dry and wet nitrogen deposition in typical agricultural areas of North Shaanxi].
To investigate the farmland soil nitrogen input from atmospheric dry and wet deposition, a 1-year observation was conducted in the Yulin and Luochuan areas of North Shaanxi Province from June 2007 to May 2008. The total inorganic nitrogen (TIN) deposition in Yulin and Luochuan was 22.17 and 16.95 kg x hm(-2) x a(-1), among which, wet deposition accounted for 95.1% and 90.4%, while dry deposition accounted for 4.9% and 9.6%, respectively, illustrating that the nitrogen deposition in both Yulin and Luochuan was mainly come from wet deposition. In the TIN deposition, the amount of nitrate in Yulin and Luochuan was 12.22 and 9.24 kg x hm(-2) xa(-1), accounting for 55.1% and 54.5%, respectively. The amount of wet deposition and the percentage of nitrate in TIN deposition were higher in Yulin than in Luochuan, because of the differences in pollution level, weather condition, and underlying surface characteristics.


Simultaneously nearly zero forward and nearly zero backward scattering objects.
With theoretical analyses and numerical calculations, we show that a passive scatterer at the sub-wavelength scale can simultaneously exhibit both nearly zero forward scattering (NZFS) and nearly zero backward scattering (NZBS). It is related to the interference of dipolar quadrupole modes of different origin, leading to coexistence of Kerker's first and second conditions at the same time. For optical frequencies, we propose two different sets of composited materials in multi-layered nano-structures, i.e., CdTe/Si/TiO2 and TiO2/Au/Si, for the experimental realization.


The biological role of the unique molecule RCAS1: a bioactive marker that induces connective tissue remodeling and lymphocyte apoptosis.
RCAS1 is a receptor-binding cancer antigen which is expressed on human uterine cervical adenocarcinoma cell line (SiSo). Finding a correlation between the expression of this gene and the overall survival of patients with 14 different types of cancer points to the clinical significance of this gene. Moreover, the expression RCAS1 correlates with other clinicopathological parameters including the histological type of cancer, its differentiation, tumor size, clinical stage, the depth of invasion, lymphovascular space involvement, lymph node metastasis, and positive peritoneal cytological results. RCAS1 can induce apoptosis in peripheral lymphocytes in vitro as well as in an increased number of apoptotic tumor-infiltrating lymphocytes. RCAS1 is also believed to contribute to the escape of tumor cells from immune surveillance. RCAS1 is secreted via ectodomain shedding, and its expression is related to changes in the characteristics of the extracellular matrix and to a reduced number of vimentin-positive tumor stromal cells, findings that suggest that RCAS1 may induce connective tissue remodeling. The concentration of RCAS1 in serum or pleural effusions has been found to be significantly higher in patients with several different types of cancer as compared to normal controls. Together, the available data shows that RCAS1 may have value as a biomarker for monitoring therapeutic efficacy. Further exploration of the biological function of RCAS1 should help in the development of new therapeutic strategies against human malignancies.


Frustrated by lack of job opportunities in Canada, MD moves to US space-medicine program.
A Canadian physician-engineer who had difficulty finding suitable work in Canada is now employed by an American company that is helping develop the medical systems that will serve astronauts. Dr. Douglas Hamilton thinks research in space medicine will have applications on earth.


[The comparative effect of continuous and intermittent maternal separation on catecholamines blood concentrations among rat pups].
Study was designed to show the affects of short-term intermittent and/or continuous maternal separation on norepinephrine and dopamine blood concentrations among rat pups. The 128 rat pups on the first, 15th, 21st, and 60th days after birth were investigated. Catheholamins blood concentration were measured by spectral fluorometry. The pathological changes in norepinephrine (NE) and dopamine (DA) plasma concentrations were exposed in both occasions - among rat pups with intermittent as well as with continuous maternal separation but were less pronounced in rat pups exposed to permanent maternal deprivation. The intermittent maternal separation produced a significant increase in plasma NE concentration compared to control non-deprived (p<0,001) and also to permanently deprived rats (p<0,01). While DA concentration, instead of elevation, was decreased in deprived rats. It was significantly lower in continuously deprived animals. The accentuated increase of NE suggests sympathetic nervous system hyperactivity, while decrease of DA indicates discoordination of the dopaminergic and sympathoadrenal systems in deprived pups. Thus, early maternal separation in rat pups may serve to be the stressful stimulus for CNS and alter it function, which produces the behavioral disorders among them.


Evaluation of 2.0-mm Titanium Three-Dimensional Curved Angle Strut Plate in the Fixation of Mandibular Angle Fractures-A Prospective Clinical and Radiological Analysis.
The aim of the study was to evaluate the efficacy and complication rate with use of 2.0-mm titanium three-dimensional (3D) curved angle strut plate for mandibular angle fractures. Twenty cases requiring internal fixation of the mandibular angle by 2.0-mm titanium 3D curved angle strut plates were evaluated. Postoperative clinical and radiological analyses were done on 1st, 2nd, 4th, and 6th weeks, which included wound healing, transmitted movements, difficulty in function and occlusion, and neurosensory changes, if any. Reasonable level of success in terms of immediate jaw function was noted in all cases. Transient inferior alveolar nerve dysfunction was observed in three cases, which recovered gradually in 2 months, and surgical site infection was observed in two cases, which resolved with appropriate course of antibiotics and wound care. Fixation of mandibular angle fractures with a 2.0-mm titanium 3D curved angle strut plate is predictable, expeditious, and has less complication rate.


Simultaneous flux and current measurement from single plant protoplasts reveals a strong link between K+ fluxes and current, but no link between Ca2+ fluxes and current.
We present a thorough calibration and verification of a combined non-invasive self-referencing microelectrode-based ion-flux measurement and whole-cell patch clamp system as a novel and powerful tool for the study of ion transport. The system is shown to be capable of revealing the movement of multiple ions across the plasma membrane of a single protoplast at multiple voltages and in complex physiologically relevant solutions. Wheat root protoplasts are patch clamped in the whole-cell configuration and current-voltage relations obtained whilst monitoring net K+ and Ca2+ flux adjacent to the membrane with ion-selective electrodes. At each voltage, net ion flux (nmol m(-2) sec(-1)) is converted to an equivalent current density (mA m(-2)) taking into account geometry and electrode efficiency, and compared with the net current density measured with the patch clamp system. Using this technique, it is demonstrated that the K+-permeable outwardly rectifying conductance (KORC) is responsible for net outward K+ movement across the plasma membrane [1:1 flux-to-current ratio (1.21 +/- 0.14 SEM, n = 15)]. Variation in the K+ flux-to-current ratio among single protoplasts suggests a heterogeneous distribution of KORC channels on the membrane surface. As a demonstration of the power of the technique we show that despite a significant Ca2+ permeability being associated with KORC (analysis of tail current reversal potentials), there is no correlation between Ca2+ flux and KORC activity. A very significant observation is that large Ca2+ fluxes are electrically silent and probably tightly coupled to compensatory charge movements. This analysis demonstrates that it is mandatory to measure flux and currents simultaneously to investigate properly Ca2+ transport mechanisms and selectivity of ion channels in general.


Serological cross-sectional studies on salmonella incidence in eight European countries: no correlation with incidence of reported cases.
Published incidence rates of human salmonella infections are mostly based on numbers of stool culture-confirmed cases reported to public health surveillance. These cases constitute only a small fraction of all cases occurring in the community. The extent of underascertainment is influenced by health care seeking behaviour and sensitivity of surveillance systems, so that reported incidence rates from different countries are not comparable. We performed serological cross-sectional studies to compare infection risks in eight European countries independent of underascertainment. A total of 6,393 sera from adults in Denmark, Finland, France, Italy, Poland, Romania, Sweden, and The Netherlands were analysed, mostly from existing serum banks collected in the years 2003 to 2008. Immunoglobulin A (IgA), IgM, and IgG against salmonella lipopolysaccharides were measured by in-house mixed ELISA. We converted antibody concentrations to estimates of infection incidence ('sero-incidence') using a Bayesian backcalculation model, based on previously studied antibody decay profiles in persons with culture-confirmed salmonella infections. We compared sero-incidence with incidence of cases reported through routine public health surveillance and with published incidence estimates derived from infection risks in Swedish travellers to those countries. Sero-incidence of salmonella infections ranged from 56 (95% credible interval 8-151) infections per 1,000 person-years in Finland to 547 (343-813) in Poland. Depending on country, sero-incidence was approximately 100 to 2,000 times higher than incidence of culture-confirmed cases reported through routine surveillance, with a trend for an inverse correlation. Sero-incidence was significantly correlated with incidence estimated from infection risks in Swedish travellers. Sero-incidence estimation is a new method to estimate and compare the incidence of salmonella infections in human populations independent of surveillance artefacts. Our results confirm that comparison of reported incidence between countries can be grossly misleading, even within the European Union. Because sero-incidence includes asymptomatic infections, it is not a direct measure of burden of illness. But, pending further validation of this novel method, it may be a promising and cost-effective way to assess infection risks and to evaluate the effectiveness of salmonella control programmes across countries or over time.


A comparison of multiple imputation methods for handling missing values in longitudinal data in the presence of a time-varying covariate with a non-linear association with time: a simulation study.
Missing data is a common problem in epidemiological studies, and is particularly prominent in longitudinal data, which involve multiple waves of data collection. Traditional multiple imputation (MI) methods (fully conditional specification (FCS) and multivariate normal imputation (MVNI)) treat repeated measurements of the same time-dependent variable as just another 'distinct' variable for imputation and therefore do not make the most of the longitudinal structure of the data. Only a few studies have explored extensions to the standard approaches to account for the temporal structure of longitudinal data. One suggestion is the two-fold fully conditional specification (two-fold FCS) algorithm, which restricts the imputation of a time-dependent variable to time blocks where the imputation model includes measurements taken at the specified and adjacent times. To date, no study has investigated the performance of two-fold FCS and standard MI methods for handling missing data in a time-varying covariate with a non-linear trajectory over time - a commonly encountered scenario in epidemiological studies. We simulated 1000 datasets of 5000 individuals based on the Longitudinal Study of Australian Children (LSAC). Three missing data mechanisms: missing completely at random (MCAR), and a weak and a strong missing at random (MAR) scenarios were used to impose missingness on body mass index (BMI) for age z-scores; a continuous time-varying exposure variable with a non-linear trajectory over time. We evaluated the performance of FCS, MVNI, and two-fold FCS for handling up to 50% of missing data when assessing the association between childhood obesity and sleep problems. The standard two-fold FCS produced slightly more biased and less precise estimates than FCS and MVNI. We observed slight improvements in bias and precision when using a time window width of two for the two-fold FCS algorithm compared to the standard width of one. We recommend the use of FCS or MVNI in a similar longitudinal setting, and when encountering convergence issues due to a large number of time points or variables with missing values, the two-fold FCS with exploration of a suitable time window.


Nickel-Catalyzed Alkylation of Amide Derivatives.
We report the catalytic alkylation of amide derivatives, which relies on the use of nonprecious metal catalysis. Amide derivatives are treated with organozinc reagents, utilizing nickel catalysis, to yield ketone products. The methodology is performed at ambient temperature and is tolerant of variation in both coupling partners. A precursor to a nanomolar glucagon receptor modulator was synthesized using the methodology, underscoring the mild nature of this chemistry and its potential utility in pharmaceutical synthesis. These studies are expected to further promote the use of amides as synthetic building blocks.


Pre- and posttreatment MR imaging in AIDS-related Kaposi sarcoma of the conjunctiva and lacrimal gland.
AIDS-related Kaposi sarcoma can affect the eye: the (bulbar) conjunctiva and lacrimal gland being rare sites of occurrence. We present a case of AIDS-related Kaposi sarcoma of the conjunctiva in which MR imaging was effective in suggesting the diagnosis and aiding therapeutic strategy. We also discuss advances in imaging of ocular and orbital malignancies presented in the recent literature.


Measuring outcomes in randomized clinical trials of insomnia treatments.
Significant efforts have been made in the last decade to develop evidence-based guidelines for the treatment of insomnia and other sleep disorders. Despite such progress, there are still no standard assessment methods to document outcome and no accepted criteria to define what should be a successful outcome in the treatment of insomnia. This paper reviews methodological and conceptual issues related to the measurement of outcomes in clinical trials of insomnia. Selected studies of behavioral and pharmacological therapies of insomnia are summarized to illustrate the types of dependent variables and assessment instruments used to document treatment efficacy. Additional outcome variables and assessment methods of potential interest are discussed, and criteria for interpreting and reporting outcomes are summarized. As most studies have relied on fairly narrow criteria to define an effective treatment, the need to broaden the scope of outcome assessment is highlighted. For instance, it is essential to document treatment efficacy beyond the simple reduction of insomnia symptoms and to incorporate additional indicators of success. Given that insomnia is associated with significant morbidity, an effective treatment should not only improve sleep parameters, but it should also produce clinically meaningful changes in daytime functioning, fatigue, mood, and quality of life. The need to evaluate outcome from multiple perspectives and to develop a core-assessment battery that would consider efficacy, clinical significance, and cost-effectiveness are discussed.


Effect of phosphate fertilizer application on phosphorus (P) losses from paddy soils in Taihu Lake Region. I. Effect of phosphate fertilizer rate on P losses from paddy soil.
A field plot study was conducted on two types of paddy soils in the Taihu Lake Region, during the rice season of year 2000 in order to assess phosphorus (P) losses by runoff and vertical leaching, which are considered the two main pathways of P movement from paddy soil into its surrounding water course. Commercial NPK compound fertilizer and single superphosphate fertilizer were applied to furnish 0, 30, 150, and 300 kg applied P ha m(-2). The experiments consisted of three replicates of each treatment in Changshu site and four replicates in Anzhen site, with a plot size of 5 x 6 m2 in a randomized block. Results revealed that the average concentration range for total P (TP) in runoff was 1.857-7.883, 1.038-5.209, 0.783-1.255 and 0.572-0.691 mg P l(-1) respectively for P300, P150, P30 and P0 in Anzhen, while it was 2.431-2.449, 1.578-1.890, 1.050-1.315 and 0.749-0.941 mg P l(-1) respectively in Changshu. In all treatments, particulate P (PP) represented a major portion of the TP lost in runoff, it was 80% in Anzhen, and it was even more (>90%) in Changshu. Phosphate fertilizer treatments significantly affected P concentrations and P loads in the runoff. The mean concentration and average seasonal TP load from the P150 plots were 1.809 mg P l(-1) and 395 g P ha m(-2) season(-1) respectively, and lower than that from the P300 plots (2.957 mg P l(-1) and 652 g P ha m(-2) season(-1)). These were obviously higher than from the P30 (0.761 mg P l(-1) and 221 g P ha m(-2) season(-1)) and P0 (0.484 mg P l(-1) and 146 g P ha m(-2) season(-1)) respectively. There was no significant difference found between the P30 and the P0 in both sites. Under usual P application rate, there were total 31.7 and 20.6 tones P removed by runoff from permeable (Anzhen site) and waterlogged (Changshu site) paddy soils in the southern Jiangsu region (major part of the TLR) in the rice season of the year 2000. But if the P application rate is unusual high, or the Olsen P in soil accumulates to above a certain level, then this could sharply increase in the future. The average concentration of molybdate reactive phosphorus (MRP) in the vertical leachate from the four different P treatments ranged from 0.058 to 0.304 mg P l(-1) in Anzhen and from 0.048 to 0.394 mg P l(-1) in Changshu. P application rate significantly affected the MRP concentration at each depth in both sites, except for the 90 cm in Anzhen. The average MRP loads during the rice season moved by vertical leaching from the four treatments ranged from 163 to 855 g P ha m(-2) season(-1) in Anzhen and 208-1,825 g P ha m(-2) season(-1) in Changshu. Vertical leachate movement does not necessarily mean that it moves towards surface water and contaminate the watercourses in this flat plain paddy soil region, it does, however, imply that P can move down from surface layers of soil to deeper levels.


Interaction of spin-labeled nicotinamide adenine dinucleotide phosphate with chicken liver fatty acid synthase.
The spatial relationships between the four reduced nicotinamide adenine dinucleotide phosphate (NADPH) binding sites on chicken liver fatty acid synthase were explored with electron paramagnetic resonance (EPR) and spin-labeled analogues of NADP+. The analogues were prepared by reaction of NADP+ with 2,2,5,5-tetramethyl-1-oxy-3-pyrroline-3-carboxylic acid, with 1,1'-carbonyldiimidazole as the coupling reagent. Several esterification products were characterized, and the interaction of the N3' ester of NADP+ with the enzyme was examined in detail. Both 1H13, 14N and 2H13, 15N spin-labels were used: the EPR spectrum was simpler, and the sensitivity greater, for the latter. The spin-labeled NADP+ is a competitive inhibitor of NADPH in fatty acid synthesis, and an EPR titration of the enzyme with the modified NADP+ indicates four identical binding sites per enzyme molecule with a dissociation constant of 124 microM in 0.1 M potassium phosphate and 1 mM ethylenediaminetetraacetic acid (pH 7.0) at 25 degrees C. The EPR spectra indicate the bound spin-label is immobilized relative to the unbound probe. No evidence for electron-electron interactions between bound spin-labels was found with the native enzyme, the enzyme dissociated into monomers, or the enzyme with the enoyl reductase sites blocked by labeling the enzyme with pyridoxal 5'-phosphate. Furthermore, the EPR spectrum of bound ligand was the same in all cases. This indicates that the bound spin-labels are at least 15 A apart, that the environment of the spin-label at all sites is similar, and that the environment is not altered by major structural changes in the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)


Endoscopic submucosal dissection for treatment of mesopharyngeal and hypopharyngeal carcinomas.
Application of the endoscopic submucosal dissection (ESD) technique, as well as elevation of the larynx in cooperation with an otolaryngologist, under general anesthesia has enabled en-block resection of early mesopharyngeal and hypopharyngeal carcinomas. The aim of this study was to retrospectively evaluate the safety of ESD and the efficacy of the elevation of the larynx for treatment of early mesopharyngeal and hypopharyngeal carcinomas. The study included 16 lesions in 13 patients who underwent ESD. To provide a sufficient working space, the larynx was elevated under direct laryngoscopy, and a tube was inserted and fixed onto the laryngeal side using the slot on the back of the laryngoscope. The median maximum diameter of the lesions was 12.5 mm (range 2 - 37 mm). The en-block resection rate was 93.8 %. Lateral surgical margins in two patients were difficult to evaluate for technical reasons. The tube could not be removed from four patients on the day of the procedure due to laryngeal edema caused by the local injection. No serious complications were observed. Oral food intake was possible after a mean of 3.3 postoperative days. With adequate intraoperative and postoperative management, ESD with elevation of the larynx may be very efficient and safe for endoscopic treatment of pharyngeal lesions.


Reduced apoptosis in term placentas from gestational diabetic pregnancies.
Gestational diabetic mellitus (GDM) pregnancies have an increased risk of macrosomic infants and large placental mass, though the mechanisms explaining each of these is uncertain. We sought to evaluate the contribution of apoptosis to placental size and the expression of glucose transporters (SLC2A) in GDM pregnancies. Maternal age and pre-pregnancy body weight were documented. Newborn weights were recorded after delivery. Placentas 37-40-week gestation from control patients (no pregnancy complication) (n = 5), or with GDM (n = 5) were weighed immediately after delivery. Villous samples (4 mm diameter) were collected and divided into specimens; one was fixed in 4% paraformaldehyde for immunostaining using terminal deoxynucleotidyl transferase dUTP-mediated nick-end labeling (TUNEL) and activated caspase-3. The other specimen was snap frozen in liquid nitrogen and stored at -80°C for active caspase-3, poly(ADP-ribose) polymerase (PARP), SLC2A1 and SLC2A3 gene expression analysis. Our results showed that maternal age and pre-pregnancy body weight were significantly higher in the GDM group when compared with those from the controls (P < 0.05). The mean neonatal birth weight and placenta weight were significantly higher in the GDM group compared with that from the controls (P < 0.05). The apoptotic index of placentas (0.05 ± 0.01 v. 0.17 ± 0.04, P < 0.04), active caspase-3 polypeptide fragments and PARP protein were significantly decreased in GDM placentas as compared with controls. Further, the level of placental SLC2A1 protein expression was ∼3-fold higher in GDM placentas. Our results suggest that reduced apoptosis in GDM placentas may contribute to increased placental tissue, which together with enhanced SLC2A1 expression, could play a role in fetal macrosomia.


[The cottage hospital model, a key to better cooperation in health care--let the cottage hospital survive!].
The cottage hospital model may be defined as an intermediary service between primary care and the general hospital. On the basis of experience and studies from Finnmark county, the northern-most county in Norway, this article makes a case for a revival of the cottage hospitals. They may improve comprehensive patient care and cooperation between care levels, to the benefit of groups of patients who often are in a squeeze between care levels: the elderly, the chronically ill, and the severely ill and dying patients. The cottage hospitals may also contribute to strengthening the chain of service in acute medicine. The professional challenges of work in a cottage hospital may attract practitioners to primary health care. We suggest that 1% of the funds set aside for ongoing national programmes for the elderly, in cancer care and mental illnesses are used for cottage hospital beds, as this may contribute to increasing the viability of these programmes. The extra cost upgrading 1,000 of a total of 27,000 nursing home beds in Norway to cottage hospital standard is estimated to be modest.


Amino acid sequence of calmodulin from wheat germ.
The complete amino acid sequence of calmodulin from wheat germ was determined by isolating and sequencing the cyanogen bromide and tryptic peptides. The protein consisted of 149 amino acid residues and its amino(N)-terminus was blocked with an acetyl group. Wheat germ calmodulin lacked tryptophan and contained 1 mol each of histidine, tyrosine, cysteine, and N epsilon-trimethyllysine residues per mol of the protein. A comparison of its amino acid sequence with that of bovine brain calmodulin indicated that there were eleven amino acid subsitutions other than amide assignments, two insertions and one deletion of amino acid residues in wheat germ calmodulin.


Measurement of the Seebeck coefficient under high pressure by dual heating.
This study presents a new method for measuring the Seebeck coefficient under high pressure in a multi-anvil apparatus. The application of a dual-heating system enables precise control of the temperature difference between both ends of the sample in a high-pressure environment. Two pairs of W-Re thermocouples were employed at both ends of the sample to monitor and control the temperature difference, and independent probes were arranged to monitor the electromotive force (emf) produced by temperature oscillation at a given target temperature. The temperature difference was controlled within 1 K during the resistivity measurements to eliminate the influence of the emf owing to a sample temperature gradient. The Seebeck measurement was successfully measured from room temperature to 1400 K and was obtained by averaging the two measured values with opposite thermal gradient directions (∼20 K). Thermoelectric properties were measured on disk-shaped p-type Si wafers with two different carrier concentrations as a reference for high Seebeck coefficients. This method is effective to determine the thermoelectric power of materials under pressure.


Safety, tolerability, pharmacokinetics and pharmacodynamics of a novel farnesoid X receptor (FXR) agonist-TQA3526 in healthy Chinese volunteers: a double-blind, randomized, placebo-controlled, dose-escalation, food effect phase I study.
Background: TQA3526 is a novel farnesoid X receptor agonist developed to treat non-alcoholic steatohepatitis (NASH) or primary biliary cholangitis (PBC). This study aimed to evaluate the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of TQA3526 in healthy Chinese patients.Methods: Healthy subjects aged 18-55 years were enrolled in this double-blinded, first-in-human, placebo-controlled single ascending dose (1, 2, 5, and 10 mg) comprising food effect investigation (10 mg) and multiple dose study (2 mg and 0.2 + 0.5 + 1 mg). Safety was assessed on the basis of adverse events. The TQA3526 concentrations were analysed in the PK study. Alkaline phosphatase (ALP), fibroblast growth factor-19 (FGF19), bile acid precursor C4 (7α-hydroxy-cholest-4-ene-3-one), cholesterol, and bile acid were selected for PD analysis.Results: TQA3526 was well tolerated, and the primary adverse drug reaction was pruritus, as expected. The exposure to TQA3526 increased in a dose-dependent manner after a single dose of 1-10 mg. The exposure was higher after food intake. A steady state was reached around 5 days, and obvious plasma accumulation of TQA3526 was observed in the multiple dose study. TQA3526 increased circulating FGF-19 and decreased C4 levels in a dose-dependent manner. ALP increased only mildly in the 2 mg multiple dose cohort.Conclusions: TQA3526 (<10 mg/day) was safe and tolerable in healthy Chinese subjects. The safety profile and PK/PD characteristics of TQA3526 support further evaluation of patients with NASH or PBC. This study was registered at https://www.chictr.org.cn/ under the identifier ChiCTR1800019570.


Exploring the acceptability and feasibility of a preconception and diabetes information app for women with pregestational diabetes: A mixed-methods study protocol.
Women with diabetes are at increased risk of adverse maternal and fetal outcomes. Preconception care can improve pregnancy outcomes and is paramount to minimise complications, but, current provision is sub-optimal. Mobile technology, particularly smartphones and apps have the potential to improve preconception care provision but research is lacking in this area. The need to use modern technologies to improve preconception care knowledge and awareness led to the development of a preconception and diabetes information app in Stage A of this study. The aim of this paper, Stage B of the study, is to explore the feasibility and acceptability of the Preconception and Diabetes Information app to improve preconception care knowledge and attitudes in women with diabetes, and explore the potential for wider implementation. A mixed-methods study design adopting a quasi-experimental approach will assess women's knowledge and attitudes related to preconception care, and level of patient activation (knowledge and confidence for self-management of health) before and after the three-month intervention period. A log of activity will be used to determine engagement with the app and semi-structured interviews will explore women's experiences. This is the first study to explore the acceptability and feasibility of a preconception and diabetes information app for women with diabetes. The app has potential to change the way preconception care is delivered, improve pregnancy outcomes and be widely implemented both in developed and developing countries. This is important given the considerable shortfalls in current preconception care services in the United Kingdom and around the world.


[Study of intermolecular interactions and self-organization of adenylic nucleotides by the spin label method].
A spin labeling method for obtaining thermodynamic parameters of nucleotide association is proposed. The method is based on the dependence of ESR parameters of the spin-labelled derivative on concentration of the nonlabelled compound due to formation of associates involving both spin-labelled and unmodified molecules. It is found that at pH 7.5 the constant of adenylic nucleotide association practically does not depend on the number of phosphate groups and is equal to 9.7 +/- 0.3 M-1 for AMP, ADP and ATP in 0.1 M NaCl at 28 degrees C. In acidic medium the value of the association constant increases by a factor two. Base stacking is shown to make the main contribution to stability of the associates of adenylic nucleotides at neutral pH, whereas upon base protonation the key role is played, apparently, by base-phosphate interaction. It is thought, that an increase in the solvent entropy is essential for stabilisation of the associates, this factor being more important in the case of nucleotide association as compared to the association of nucleosides. A possible role of nucleotide association in the processes of intracellular regulation is discussed.


Prevalence of taking actions to control blood pressure among adults with self-reported hypertension in 18 states and the District of Columbia, 2009.
The authors used 2009 Behavioral Risk Factor Surveillance System data to assess the prevalence of taking actions to control hypertension among adults with self-reported hypertension. Differences by descriptive characteristics (sex, age, race/ethnicity, access to health care, medication adherence), presence of other health risk factors (overweight/obesity, smoking, heavy drinking, inadequate fruit/vegetable intake, and physical inactivity), and comorbidities (diabetes, high cholesterol, coronary heart disease, and stroke) were compared. The prevalence of hypertension was 29.6%, and 75.0% of these patients reported taking antihypertensive medications, 73.1% changed eating habits, 72.8% decreased the use of salt, 78.8% reduced alcohol consumption, and 69.9% increased their physical activity. Overall, 87.2% reported taking two or more actions to reduce blood pressure. Patients taking antihypertensive medications were more likely to take two or more actions than their counterparts (90.6% vs 79.4%, P<.01). Those with at least one other health risk factor were 1.85 times as likely to take two or more actions as their counterparts (95% confidence interval, 1.18-2.92 times). More than 80% of hypertensive adults reported taking two or more actions to control blood pressure. The prevalence of taking actions differed significantly by descriptive characteristics, the presence health risk factors, and comorbidities.


MDR in cancer: Addressing the underlying cellular alterations with the use of nanocarriers.
Multidrug resistance (MDR) is associated with a wide range of pathological changes at different cellular and intracellular levels. Nanoparticles (NPs) have been extensively exploited as the carriers of MDR reversing payloads to resistant tumor cells. However, when properly formulated in terms of chemical composition and physicochemical properties, NPs can serve as beyond delivery systems and help overcome MDR even without carrying a load of chemosensitizers or MDR reversing molecular cargos. Whether serving as drug carriers or beyond, a wise design of the nanoparticulate systems to overcome the cellular and intracellular alterations underlying the resistance is imperative. Within the current review, we will initially discuss the cellular changes occurring in resistant cells and how such changes lead to chemotherapy failure and cancer cell survival. We will then focus on different mechanisms through which nanosystems with appropriate chemical composition and physicochemical properties can serve as MDR reversing units at different cellular and intracellular levels according to the changes that underlie the resistance. Finally, we will conclude by discussing logical grounds for a wise and rational design of MDR reversing nanoparticulate systems to improve the cancer therapeutic approaches.


Bisulfite sequencing Data Presentation and Compilation (BDPC) web server--a useful tool for DNA methylation analysis.
During bisulfite genomic sequencing projects large amount of data are generated. The Bisulfite sequencing Data Presentation and Compilation (BDPC) web interface (http://biochem.jacobs-university.de/BDPC/) automatically analyzes bisulfite datasets prepared using the BiQ Analyzer. BDPC provides the following output: (i) MS-Excel compatible files compiling for each PCR product (a) the average methylation level, the number of clones analyzed and the percentage of CG sites analyzed (which is an indicator of data quality), (b) the methylation level observed at each CG site and (c) the methylation level of each clone. (ii) A methylation overview table compiling the methylation of all amplicons in all tissues. (iii) Publication grade figures in PNG format showing the methylation pattern for each PCR product embedded in an HMTL file summarizing the methylation data, the DNA sequence and some basic statistics. (iv) A summary file compiling the methylation pattern of different tissues, which is linked to the individual HTML result files, and can be directly used for presentation of the data in the internet. (v) A condensed file, containing all primary data in simplified format for further downstream data analysis and (vi) a custom track file for display of the results in the UCSC genome browser.


Increased divergence but reduced variation on the Z chromosome relative to autosomes in Ficedula flycatchers: differential introgression or the faster-Z effect?
Recent multilocus studies of congeneric birds have shown a pattern of elevated interspecific divergence on the Z chromosome compared to the autosomes. In contrast, intraspecifically, birds exhibit less polymorphism on the Z chromosome relative to the autosomes. We show that the four black-and-white Ficedula flycatcher species show greater genetic divergence on the Z chromosome than on the autosomes, and that the ratios of intraspecific polymorphism at Z-linked versus autosomal markers are below the neutral expectation of 75%. In all species pairs, we found more fixed substitutions and fewer shared polymorphisms on the Z chromosome than on the autosomes. Finally, using isolation with migration (IMa) models we estimated gene flow among the four closely related flycatcher species. The results suggest that different pattern of evolution of Z chromosomes and autosomes is best explained by the faster-Z hypothesis, since the estimated long-term gene flow parameters were close to zero in all comparisons.


Handheld ultrasound to reduce requests for inappropriate echocardiogram (HURRIE).
Handheld ultrasound could provide sufficient information to satisfy the clinical questions underlying 'rarely appropriate' echo requests, but there are limited data about its use as a gatekeeper to standard echocardiography. We sought to determine whether the use of handheld ultrasound could improve the appropriate use of echocardiography. A prospective study comparing handheld ultrasound strategy to standard echocardiography for studies deemed rarely appropriate, using a questionnaire based on appropriate use criteria was conducted across two hospitals, from October 2017 to April 2018. Groups undergoing Handheld ultrasound (n = 76, 58 (46.5-72.5) years, 53 males, 78% outpatients) and standard echocardiography (n = 72, 61 (49.0-71.5) years, 42 males, 76% outpatients) were comparable. There was a significant decrease in the time to scan from just over 1 month in standard group to a median of 12 days in handheld ultrasound group (P < 0.001). This difference was small for inpatients (from 1 day to a median of 10 min in handheld ultrasound, P = 0.014), but prominent in outpatients (from 1.5 months in the standard group to median of 2 weeks in the handheld ultrasound group, P < 0.001). There was no increase in the need for follow-up scan within 6 months and no significant differences in length of hospital stay for inpatients. Handheld ultrasound can be an effective gatekeeper to standard echocardiography for requests deemed rarely appropriate, reducing time to echocardiography significantly and potentially decreasing the need for standard echocardiography by up to 20%.


Should public health interventions aimed at reducing childhood overweight and obesity be gender-focused?
Overweight in childhood is a major public health concern that calls for immediate preventative action. An increasing number of reports suggest that gender specific approaches to prevention may be more effective. However, there is a paucity of information to guide gender-sensitive health promotion and population health interventions for the prevention of overweight in childhood. In the present study, we sought to determine gender-differentials in overweight and underlying behaviors, nutrition and physical activity, among pre-adolescents in Alberta, Canada, to inform the discussion on gender-focused interventions for chronic disease prevention. In 2008, we surveyed 3421 grade five students and their parents of 148 randomly selected schools. Students completed the Harvard food frequency questionnaire, questions on physical activities, and had their height and weight measured. Parents completed questions on socio-economic background and child's lifestyle. We applied multilevel regression methods to assess gender differentials in overweight, nutrition and physical activity. Overall, the prevalence of overweight was slightly higher among boys (29.1%) than girls (27.9%) with more pronounced differences in towns and urban geographies. Boys reported to be much more physically active relative to girls (OR = 2.12, 95% CI: 1.73-2.60). Diets of boys, relative to those of girls, reportedly constituted more fat and were less likely to meet the recommendation of 6 daily servings of vegetables and fruits (OR = 0.81, 95% CI: 0.71-0.93). Our findings confirm the existence of gender differences in physical activity and nutrition, and support gender-focused health promotion whereby priority is given to physical activity among girls and to healthy eating among boys.


Post-Traumatic Stress Disorder (Ptsd) Following Burn Injuries: A Comprehensive Clinical Review.
Burns are a global public health problem with a large number of psychosocial and psychological problems that may ensue after burn injuries. One of the commonest psychological problems seen after burn injuries is depression, anxiety and post-traumatic stress disorder (PTSD). The following paper reviews the existing literature on PTSD following burn injuries. The concept of PTSD is explained for the non-psychiatrist involved in burn rehabilitation. The various psychosocial factors that determine the causation and course of PTSD following burn injuries are discussed. PTSD following burn injuries in special populations like women, children and adolescents, intentional burns and self immolation are also discussed. The role of cultural factors in the development of PTSD are elucidated and future research needs are laid out. PTSD is a very common occurrence after burn injuries and needs a multidisciplinary team evaluation for its management.


[Evaluation of a new corticoid for various dermatoses].
Diflucortolone valerate 0.1% (investigational number SH K 183 MB) in the form of a cream was clinically investigated in 41 patients with dermatological disorders. All the patients had been treated previously with other topical preparations, but without success. Complete healing or distinct improvement of the objective and subjective symptoms of the disorders was achieved in 100% of the cases treated with the investigational preparation. No side effects were observed at all.


Purification and characterization of a major glycoprotein in rat hepatoma plasma membranes. One of the membrane proteins released by phosphatidylinositol-specific phospholipase C.
A major glycoprotein of rat hepatoma plasma membranes was selectively released as a soluble form by incubating the membrane with phosphatidylinositol-specific phospholipase C. The soluble form corresponding to the glycoprotein was also prepared by butan-1-ol extraction of microsomal membranes at pH 5.5, whereas extraction at pH 8.5 yielded an electrophoretically different form with a hydrophobic nature. The soluble glycoprotein extracted at pH 5.5 was purified by sequential chromatography on concanavalin A-Sepharose, Sephacryl S-300 and anti-(alkaline phosphatase) IgG-Sepharose, the last step being used to remove a contaminating alkaline phosphatase. The glycoprotein thus purified was a single protein with Mr 130,000 in SDS/polyacrylamide-gel electrophoresis, although it behaved as a dimer in gel filtration on Sephacryl S-300. The glycoprotein was analysed for amino acid and carbohydrate composition. The composition of the carbohydrate moiety, which amounted to 64% by weight, suggested that the glycoprotein contained much larger numbers of N-linked oligosaccharide chains than those with O-linkage. It was confirmed that the purified glycoprotein was immunologically identical not only with that released by the phospholipase C but also with the hydrophobic form extracted with butan-1-ol at pH 8.5. The results indicate that the glycoprotein of rat hepatoma plasma membranes, which has an unusually high content of carbohydrate, is another membrane protein released by phosphatidylinositol-specific phospholipase C, as documented for alkaline phosphatase, acetylcholinesterase and Thy-1 antigen.


Using scientific evidence to inform public policy on the long distance transportation of animals: role of the European Food Safety Authority.
The authors review the work of the previous Scientific Committee on Animal Health and Animal Welfare and the current European Food Safety Authority (EFSA) in providing scientific advice on the welfare aspects of animal transport and the impact of this advice on the European Union (EU) regulatory framework. Through its Protocol on the Protection and Welfare of Animals, the Treaty of Amsterdam obliges European institutions to pay full regard to the welfare requirements of animals when formulating and implementing EU legislation. Regulation 1/2005 states that EU legislation should be amended to take into account new scientific evidence. Provisions for poultry, cats and dogs take into account the recommendations included in EFSA's Scientific Opinion which considers different species (poultry, deer, rabbits, dogs and cats, fish and exotic animals). Examples of the effect of the scientifically based conclusions and recommendations from the Scientific Opinion on the measures in Regulation 1/2005 are summarised and show the impact of scientific evidence on EU legislation.


Primary Care for Mentally Ill Adults in Acute Residential Treatment Facilities.
Psychiatric bed capacity has decreased nationally with an impact on the number and the types of patients admitted to acute in-patient units. Acute residential programs provide alternatives to hospitals for patients with problems that do not meet criteria for admission. However, these settings may not have resources to respond to the medical co-morbidities associated with severe mental illness. To understand the medical needs of patients in these settings, this research examined data from a nurse-managed primary care outreach service. Prevalent problems included acute issues and chronic disorders associated with severe mental illness. In multivariate analyses, demographic and substance related factors contributed to all chronic illness. However, age was the over-riding predictor of cardio-metabolic disorders. Drug abuse consistently predicted HIV/AIDS and HCV risk. Findings demonstrate that patients diverted to alternative forms of acute psychiatric care are no less in need of medical services than those admitted to hospitals. This underscores the importance of primary care that is provided at the site of acute residential treatment and integrated with overall care.


Characterization of mutations in streptomycin-resistant Mycobacterium tuberculosis clinical isolates in the area of Barcelona.
To determine the proportion and type of mutations in Mycobacterium tuberculosis isolates resistant to streptomycin, and their relationship with the level of resistance and with the epidemiological molecular pattern of the isolates. Sixty-nine streptomycin-resistant isolates from a M. tuberculosis strain collection (1995-2005) from Barcelona were studied. The MIC of streptomycin for each isolate was determined using the proportions method with Middlebrook 7H11 medium. The entire rpsL gene and two specific fragments of the rrs gene (the 530 loop and the 912 region) were sequenced. IS6110-restriction fragment length polymorphism and spoligotyping were performed in each isolate. Twenty-six (26/69, 37.7%) streptomycin-resistant isolates presented a mutation in either the rpsL gene and/or the rrs530 loop, with no mutation in the rrs912 region. Seventeen (24.6%) isolates showed rpsL mutations (codons 43 and 88) associated with high MIC levels. Nine (13.0%) isolates had alterations in the rrs gene (A513T, A513C and C516T). Nineteen isolates (19/64, 29.7%) were classified into seven clusters (containing 2-5 isolates per cluster). Nineteen different spoligotype patterns were found. All the LAM3 spoligotype isolates (10/67, 14.9%) were associated with a C491T change in the rrs gene, being also observed in all LAM3 streptomycin-susceptible isolates. Mutations in the rpsL and rrs genes were detected in 37.7% of streptomycin-resistant M. tuberculosis isolates. High-level resistance was associated with mutations in the rpsL gene, whereas wild-type isolates showed low MIC levels. The presence of the C491T substitution in the rrs gene in streptomycin-susceptible and -resistant isolates demonstrates that this change is an epidemiological marker associated with LAM3 sublineage.


The Effect of Glyphosate on Human Sperm Motility and Sperm DNA Fragmentation.
Glyphosate is the active ingredient of Roundup®, which is one of the most popular herbicides worldwide. Although many studies have focused on the reproductive toxicity of glyphosate or glyphosate-based herbicides, the majority of them have concluded that the effect of the specific herbicide is negligible, while only a few studies indicate the male reproductive toxicity of glyphosate alone. The aim of the present study was to investigate the effect of 0.36 mg/L glyphosate on sperm motility and sperm DNA fragmentation (SDF). Thirty healthy men volunteered to undergo semen analysis for the purpose of the study. Sperm motility was calculated according to WHO 2010 guidelines at collection time (zero time) and 1 h post-treatment with glyphosate. Sperm DNA fragmentation was evaluated with Halosperm® G2 kit for both the control and glyphosate-treated sperm samples. Sperm progressive motility of glyphosate-treated samples was significantly reduced after 1 h post-treatment in comparison to the respective controls, in contrast to the SDF of glyphosate-treated samples, which was comparable to the respective controls. Conclusively, under these in vitro conditions, at high concentrations that greatly exceed environmental exposures, glyphosate exerts toxic effects on sperm progressive motility but not on sperm DNA integrity, meaning that the toxic effect is limited only to motility, at least in the first hour.


Smoking rates among American Indian women giving birth in Minnesota. A call to action.
Maternal smoking is associated with a number of adverse effects including neonatal mortality, preterm delivery, low birth weight, decreased lung growth and function, and increased incidence of lower respiratory infections. This study looked at smoking trends among American Indian and white women in Minnesota who gave birth between 1999 and 2003. The authors used data from birth certificates issued by the state to determine the rate of smoking among both groups of women. They broke down the data further to look at and compare smoking rates of American Indian and white women based on where they gave birth (greater Minnesota or the Twin Cities metro area), when they began receiving prenatal care, their level of education, their age, the number of children they already had, and the weight of their baby. They found that 39% of American Indian women in Minnesota who gave birth during that period reported smoking, compared with 11% of white women. In addition, their findings showed that cigarette use among American Indian women in Minnesota who gave birth in 2003, was 36.5%--double the national rate of 18.3% forAmerican Indian women.


Mutations in PMR5 result in powdery mildew resistance and altered cell wall composition.
Powdery mildews and other obligate biotrophic pathogens are highly adapted to their hosts and often show limited host ranges. One facet of such host specialization is likely to be penetration of the host cell wall, a major barrier to infection. A mutation in the pmr5 gene rendered Arabidopsis resistant to the powdery mildew species Erysiphe cichoracearum and Erysiphe orontii, but not to the unrelated pathogens Pseudomonas syringae or Peronospora parasitica. PMR5 belongs to a large family of plant-specific genes of unknown function. pmr5-mediated resistance did not require signaling through either the salicylic acid or jasmonic acid/ethylene defense pathways, suggesting resistance in this mutant may be due either to the loss of a susceptibility factor or to the activation of a novel form of defense. Based on Fourier transform infrared analysis, the pmr5 cell walls were enriched in pectin and exhibited a reduced degree of pectin modification relative to wild-type cell walls. In addition, the mutant had smaller cells, suggesting a defect in cell expansion. A double mutant with pmr6 (defective in a glycosylphosphatidylinositol-anchored pectate lyase-like gene) exhibited a strong increase in total uronic acid content and a more severe reduction in size, relative to the single mutants, suggesting that the two genes affect pectin composition, either directly or indirectly, via different mechanisms. These two mutants highlight the importance of the host cell wall in plant-microbe interactions.


Immunological relationships among the myeloma proteins.
An immunological study of 21 myeloma sera was carried out to determine their relationship to components of normal gamma-globulin and to each other. Ten proteins were separated for detailed characterization. Every one of the myeloma proteins studied was immunologically different, indicating individual specificity. The gamma-type myeloma proteins were all related to normal gamma-globulin or a fraction thereof, and although each was distinct from the others, all were related. The beta-type myeloma proteins likewise were related to one another, though individually specific. Evidence was found favoring the view that some of the gamma-type myeloma proteins are related to the beta-type myeloma proteins although most of them were completely unrelated. Likewise the results indicated a definite but distant relationship between the beta-myeloma proteins and normal gamma-globulin. Certain of the beta-myeloma proteins were more closely related to a fraction of normal serum of similar electrophoretic mobility. It is suggested that most multiple myeloma proteins fall into two groups or families of cross-reacting abnormal proteins, one closely related to normal gamma-globulin, the other distantly related to gamma-globulin with marked individual specificities.


Positive facial expressions are recognized faster than negative facial expressions, but why?
Three experiments examined the recognition speed advantage for happy faces. The results replicated earlier findings by showing that positive (happy) facial expressions were recognized faster than negative (disgusted or sad) facial expressions (Experiments 1 and 2). In addition, the results showed that this effect was evident even when low-level physical differences between positive and negative faces were controlled by using schematic faces (Experiment 2), and that the effect was not attributable to an artifact arising from facilitated recognition of a single feature in the happy faces (up-turned mouth line, Experiment 3). Together, these results suggest that the happy face advantage may reflect a higher-level asymmetry in the recognition and categorization of emotionally positive and negative signals.


Characterization of [3H]ethynylbicycloorthobenzoate ([3H]EBOB) binding and the action of insecticides on the gamma-aminobutyric acid-gated chloride channel in cultured cerebellar granule neurons.
[3H]Ethynylbicycloorthobenzoate ([3H]EBOB) is a recently developed specific radioligand for the noncompetitive blocker site of the gamma-aminobutyric acid (GABA) receptor, which is the target of many polychlorocycloalkane and heterocyclic insecticides. This study characterizes [3H]EBOB binding and the action of insecticides on the GABA-gated chloride channel of cerebellar granule neurons in primary culture. Specific [3H]EBOB binding increases rapidly between days 3 and 7 and then remains constant for days 7 to 12 in culture. A single, high-affinity and saturable binding site is involved with Kd = 0.51 nM and Bmax = 239 fmol/mg protein. GABA and muscimol are 6- to 12-fold more potent as inhibitors of [3H]EBOB binding to membranes from cultured cerebellar granule neurons than from cerebellar tissue, possibly because of localization of specific subunits in the granule cells. GABA-stimulated 36Cl- influx is also inhibited by seven insecticidal compounds (including EBOB, 12-ketoendrin and alpha-endosulfan) as well as picrotoxinin and t-butylbicyclophosphorothionate with IC50 levels ranging from 290 to 15300 nM, which correlate well (r = 0.92) with their inhibitory potencies for [3H]EBOB binding (1 to 773 nM). These findings validate the use of [3H]EBOB binding in cultured cerebellar granule cells for pharmacological studies on insecticides and other toxicants acting at the noncompetitive blocker or coupled site of the GABAA receptor/chloride channel complex.


Intracellular localization of alpha-tocopherol transfer protein and alpha-tocopherol.
The mechanism of action of tocopherol transfer protein (TTP) and its role in the intracellular processing of vitamin E were investigated using confocal fluorescence microscopy. The results from this work suggest that TTP functions by transporting vitamin E from endocytic organelles to other locations in the cell.


Atriopeptin III. A potent natriuretic, diuretic, and hypotensive agent in rats with chronic renal failure.
Chronic renal failure is frequently associated with volume overload, resulting in hypertension and, in some cases, congestive heart failure. Atriopeptin III (AP III), a 24-amino acid atrial peptide, is a potent vasodilator and natriuretic/diuretic agent in normal rats. An infusion of AP III at 0.2 microgram/kg per min for 60 min produced dramatic responses in animals with chronic renal failure (5/6 nephrectomy 4 wk before study). Systemic blood pressure fell 20% by the end of infusion. A pronounced rise in glomerular filtration rate (24%) was maintained during the infusion period when urine flow rate was stable (35-60 min), even though renal blood flow was unchanged from base line. Urinary volume increased 4.4-fold and sodium excretion increased 9 to 12-fold during the infusion. Fractional excretion of sodium ranged between 9 and 15% in those animals whose initial GFR values were lower than 0.5 ml/min. We conclude that AP III is a potent natriuretic/diuretic agent in rats with reduced renal mass, presumably exerting that effect predominantly through increases in GFR. This agent may well be useful in the treatment of volume overload in patients with chronic renal failure.


Aberrant link between empathy and social attribution style in borderline personality disorder.
In social interactions, we often need to quickly infer why other people do what they do. More often than not, we infer that behavior is a result of personality rather than circumstances. It is unclear how the tendency itself may contribute to psychopathology and interpersonal dysfunction. Borderline personality disorder (BPD) is characterized by severe interpersonal dysfunction. Here, we investigated if this dysfunction is related to the tendency to over-attribute behaviors to personality traits. Healthy controls and patients with BPD judged positive and negative behaviors presented within a situational constraint during functional magnetic resonance imaging. Before the experiment, we measured trait levels of empathy, paranoia, and need for cognition. Behaviorally, we found that empathy levels predicted the tendency to attribute behavior to traits in healthy controls, whereas in patients with BPD this relationship was significantly weakened. Whole brain analysis of group-by-empathy interaction revealed that when participants judged the behavior during the attribution phase, several brain regions implicated in mentalizing distinguished patients from controls: In healthy controls, neural activity scaled negatively with empathy, but this relationship was reversed in BPD patients. Due to the cross-sectional study design we cannot establish a causal link between empathy and social attributions. These findings indicate that the self-reported tendency to feel for others is related to the tendency to integrate situational information beyond personality. In BPD patients, by contrast, the association between empathy and attribution was significantly weaker, rendering empathy less informative in predicting the overall attribution style.


Guamanian neurodegenerative disease: electrophysiologic findings.
Amyotrophic lateral sclerosis (ALS), parkinsonism and/or dementia are highly prevalent among the Chamorro population of Guam. The incidence of Guamanian ALS has markedly declined in recent years, but these incidence figures may reflect underascertainment of subclinical disease. Guamanian Chamorro patients have not been systematically studied using modern clinical neurophysiological techniques. Electromyography (EMG: needle exam and nerve conduction studies) was used to study 29 patients with the major subtypes of Guamanian neurodegenerative disease, as well as 11 neurologically normal Guamanian Chamorro subjects. Central conduction was assessed by somatosensory evoked potentials (SEP's) in 16 patients. EMG evidence of peripheral neuropathy, (often subclinical) was found in 45% of Guamanian patients but no Chamorro control subjects. Diabetes mellitus, which is highly prevalent in this population, was present in some, but not all of these cases. Clinically unsuspected motor neuron disease was identified by EMG in only one of the 23 Guamanian patients with parkinsonism and/or dementia and in none of the 11 Chamorro control subjects. Two of seven patients with the clinical phenotype of Guamanian ALS had a more benign EMG pattern on the needle electrode exam with absence of fibrillation and fasciculation potentials. Three of 16 patients (all with parkinsonism and dementia) had mildly abnormal tibial SEP's. No patient had EMG evidence of myopathy or a defect of neuromuscular transmission. We conclude: (1) peripheral neuropathy may be a manifestation of Guamanian neurodegenerative disease; (2) the declining prevalence of ALS on Guam is not associated with the development of a subclinical form of motor neuron disease; (3) the substantial overlap of Guamanian ALS with parkinsonism-dementia reported in prior decades is no longer apparent; (4) abnormal central conduction, as assessed by tibial SEP's, is present in some patients with Guamanian parkinsonism-dementia.


Effective wedge angles with a universal wedge.
Some recently designed x-ray-producing accelerators are equipped with a single built-in wedge, and different 'effective' wedge angles are obtained by combining an open (unwedged) and a wedged field in the appropriate proportions. This paper describes a technique for determining these proportions from measured isodose distributions of the two component fields. Our data for the Philips SL/75 6 MV accelerator are compared with two existing theoretical models. One model, in which the beams are weighted by the ratio of the tangents of the effective and nominal wedge angles, agrees with the data to within 3 degrees over the range of effective wedge angles and square field sizes examined. The second and simpler model, in which the beams are weighted by the ratio of the wedge angles directly, results in errors of as much as 11 degrees. It is shown that both of these models are approximations to an exact theoretical solution which may be formulated in terms of one free parameter. This parameter may be interpreted physically as the ratio of the slopes of the central-axis depth-dose curves for the open and wedged fields.


Severe acute respiratory syndrome coronavirus 2 prevalence in saliva and gastric and intestinal fluid in patients undergoing gastrointestinal endoscopy in coronavirus disease 2019 endemic areas: Prospective cross-sectional study in Japan.
Gastrointestinal endoscopy (GIE) is useful for the early detection and treatment of many diseases; however, GIE is considered a high-risk procedure in the coronavirus disease 2019 (COVID-19) pandemic era. This study aimed to explore the rate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positivity in saliva and gastrointestinal fluids to which endoscopy medical staff are exposed. The study was a single-center cross-sectional study. From June 1 to July 31, 2020, all patients who underwent GIE at Yokohama City University Hospital were registered. All patients provided 3 mL of saliva. For upper GIE, 10 mL of gastric fluid was collected through the endoscope. For lower GIE, 10 mL of intestinal fluid was collected through the endoscope. The primary outcome was the positive rate of SARS-CoV-2 in saliva and gastrointestinal fluids. We also analyzed serum-specific antibodies for SARS-CoV-2 and patients' background information. A total of 783 samples (560 upper GIE and 223 lower GIE samples) were analyzed. Polymerase chain reaction (PCR) on saliva samples did not show any positive results in either upper or lower GIE samples. However, 2.0% (16/783) of gastrointestinal fluid samples tested positive for SARS-CoV-2. No significant differences in age, sex, purpose of endoscopy, medication, or rate of antibody test positivity were found between PCR positive and PCR negative cases. Asymptomatic patients, even those with no detectable virus in their saliva, had SARS-CoV-2 in their gastrointestinal tract. Endoscopy medical staff should be aware of infection when performing procedures. The study was registered as UMIN000040587.


Anatomical variants in the anterosuperior aspect of the glenoid labrum: a statistical analysis of seventy-three cases.
Anterosuperior labral variations of the glenohumeral joint have been previously described and are thought to represent normal anatomical variants without any known clinical importance. The goals of this study were to characterize anterosuperior labral anatomical variations and to determine their prevalence and clinical importance. A total of 546 patients who had an arthroscopic procedure in the shoulder were prospectively evaluated for variations of the anterosuperior aspect of the labrum. Structural characteristics of the anatomical variants were noted and recorded intraoperatively. The relationships of these anterosuperior labral variations to demographic data, preoperative symptoms, findings on physical examination, and associated intra-articular abnormalities were analyzed statistically and compared with findings in patients with normal anterosuperior labral anatomy. Three distinct variations of the anterosuperior portion of the labrum were found in seventy-three patients (13.4%): a sublabral foramen only (eighteen patients; 3.3%), a sublabral foramen with a cord-like middle glenohumeral ligament (forty-seven; 8.6%), and an absence of labral tissue at the anterosuperior portion of the labrum with a cord-like middle glenohumeral ligament (eight; 1.5%). In multivariate analysis, the presence of one of these three variations revealed a significant positive association with anterosuperior labral fraying (p = 0.000), an abnormal superior glenohumeral ligament (p = 0.012), and increased passive internal rotation with the arm in 90 degrees of abduction (p = 0.046). These lesions were negatively associated with a painful arc sign and the presence of either a partial or full-thickness supraspinatus tendon tear. Statistical analysis revealed a clinical difference between the cord-like middle glenohumeral ligament variants and the sublabral foramen variant. Anterosuperior labral variants may influence glenohumeral biomechanics and may predispose the shoulder to other abnormalities. These variants are associated with increased internal rotation, which may predispose the shoulder to lesions of the superior glenohumeral ligament and anterosuperior portion of the labrum. These variations themselves do not appear to contribute to instability.


Dysfunction of DNA damage-inducible transcript 4 in the decidua is relevant to the pathogenesis of preeclampsia.
Preeclampsia (PE) is a pregnancy-related disorder that occurs after 20 weeks of gestation and affects 3-5% of all human pregnancies worldwide. However, the pathogenesis of PE still remains poorly understood. A deficiency in decidualization is considered a contributing factor to the development of PE. The DNA damage inducible transcript 4 (DDIT4) gene encodes a protein whose main function is inhibiting mammalian target of rapamycin (mTOR) under stress, and several studies have demonstrated that its expression promotes tumor cell apoptosis. Our previous RNA-Seq results showed that DDIT4 is significantly decreased in the decidua of PE women. Here, we aimed to define the role of DDIT4 in human decidualization and its relationship with PE. The results indicated that DDIT4 was markedly decreased in the decidua of severe PE compared with those from uncomplicated pregnancies. The expression of DDIT4 in human endometrial stromal cell (hESC) line and primary hESCs was up-regulated during decidualization. Knockdown DDIT4 in hESCs and primary hESCs caused a significant reduction in the transcription of decidualization markers, insulin-like growth factor binding protein 1 (IGFBP1) and prolactin (PRL). In addition, silencing DDIT4 caused up-regulated p-mTOR and p-p70s6k and reduced apoptosis, whereas rapamycin, an inhibitor of mTOR, reversed the result of apoptosis. Moreover, the expression of cleaved-caspase 3 in severe PE was significantly lower than that of uncomplicated pregnancies, which was unfavorable for trophoblast invasion. Our data suggest that DDIT4 is critical for normal decidualization and the apoptosis of decidual cells. DDIT4 deficiency is likely involved in the development of PE.


Simultaneous multiple-modality therapy for tension headaches and neck pain.
Eighty-one patients suffering from neck pain and tension-type headaches were treated by the application of a unique physical therapy device combining transcutaneous electrical nerve stimulation (TENS), traction, massage, vibration, and acupressure applied to the forehead, posterior cervical spine, and scapula. The device employed eight silver silicone electrodes for modality application. Its safety and effectiveness were assessed by evaluating patients before and after treatment. No complication ensued. Statistical analysis of visual analog scales for neck pain and headache yielded p values < 0.0001. Analysis of fibromyalgia neck and shoulder trigger points with three controls gave nonsignificant results. Conclusions were that 1) the device is safe and effective as judged by subjective patient input and 2) fibromyalgia trigger points are unaffected by the treatments. More objective measures are needed to provide evidence of efficacy.


Occurrence and Practices for Pain, Agitation, and Delirium in Intensive Care Unit Patients.
Our study described the occurrence, assessment, prevention, and management practices of pain, agitation, and delirium (PAD) in four intensive care units (ICUs) from the Puerto Rico Medical Center and compared findings with the 2013 PAD guidelines. A descriptive study, with repeated bedside measures (two times a day/two times a week) of PAD and review of patient clinical records. Eighty ICU patients (20 per ICU) were evaluated, (median 3 times [IQR, 2-7]). At least once during the assessment period, 57% percent of patients had significant pain and 34% had delirium. Moreover, 46% were deeply sedated, 17.5% had agitation, and 52.5% of patients were within the recommended Richmond Agitation-Sedation Scale (RASS) scores. The Numeric Rating Scale and RASS were the most common tools used by clinicians to evaluate pain and agitation/sedation levels, respectively. Clinicians did not assess pain in patients unable to self-report with any guideline-recommended tools, as was the case for delirium. Fentanyl and morphine were the most commonly used analgesics, while benzodiazepines were used for sedation. Although pain, agitation, and delirium occurrence were similar to other studies, patients continue to suffer. A gap exists between clinical practices in these ICUs and current guidelines. Strategies that contribute to integrating guidelines into these ICUs should be developed, studied, and implemented.


Molecular dissection of a viral quasispecies under mutagenic treatment: positive correlation between fitness loss and mutational load.
Low fidelity replication and the absence of error-repair activities in RNA viruses result in complex and adaptable ensembles of related genomes in the viral population, termed quasispecies, with important implications for natural infections. Theoretical predictions suggested that elevated replication error rates in RNA viruses might be near to a maximum compatible with viral viability. This fact encouraged the use of mutagenic nucleosides as a new antiviral strategy to induce viral extinction through increased replication error rates. Despite extensive evidence of lethal mutagenesis of RNA viruses by different mutagenic compounds, a detailed picture of the infectivity of individual genomes and its relationship with the mutations accumulated is lacking. Here, we report a molecular analysis of a foot-and-mouth disease virus population previously subjected to heavy mutagenesis to determine whether a correlation between increased mutagenesis and decreased fitness existed. Plaque-purified viruses isolated from a ribavirin-treated quasispecies presented decreases of up to 200-fold in infectivity relative to clones in the reference population, associated with an overall eightfold increase in the mutation frequency. This observation suggests that individual infectious genomes of a quasispecies subjected to increased mutagenesis lose infectivity by their continuous mutagenic 'poisoning'. These results support the lethal defection model of virus extinction and the practical use of chemical mutagens as antiviral treatment. Even when extinction is not achieved, mutagenesis can decrease the infectivity of surviving virus, and facilitate their clearance by host immune responses or complementing antiviral approaches.


Light- and electron microscopic observations on parathyroid glands in different age groups of rats.
The parathyroid hormone (PTH) acts on bones, intestines, and kidneys to maintain the calcium homeostasis which, in turn, is a main factor in controling the parathyroid (PT) gland activity. In all mammals studied, the chief cells of PT glands changed their size, shape, and cytoplasmic structure due to different functional states which vary the serum calcium levels. The chief cells of the rat PT glands were classified as dark and light. The dark cells may constitute an active form, characterized mainly by the abundant free ribosomes, conspicuous rough endoplasmic reticulum, and GOLGI complexes, greater number of secretory granules (SG) and increased tortuosity of the plasma membranes as compared to the light ones which were considered as a less active type of cells. Due to different calcium requirements in newborn and young rats for the ossification of growing skeleton and in adult and senile rats with consolidate mature bones, the PT glands studied with electron microscope showed various cytological features. The parenchyma of newborn and young PT glands was composed by dark chief cells. The light chief cells were more frequent in adult and senile animals as a less active type of cell. Mature SG were only occasionally observed in dark cells of newborn, young and adult PT glands. They may constitute a reserve supply of PTH but probably not the main way of secretion, according to their little number. Another pool of PTH probably answers the needs for the small basal variations in the steady-state secretion and may be represented by the vesicles observed in the chief cells cytoplasm.


Neurotropic melanoma of the face invading the maxillary nerve.
Neurotropic melanoma is a variant of desmoplastic melanoma which, in addition to a spindle-cell vertical component and fibrosis, exhibits a fascicular growth pattern and invades cutaneous nerves. Here we describe a neurotropic melanoma which arose in a small lentigo maligna melanoma and invaded the maxillary nerve for a distance of 5 cm. Ultrastructural features were similar to those of previously described neurotropic melanomas and tumor fascicles contained S-100 protein and neuron-specific enolase immunoreactivity.


Early correction of malocclusion using planas direct tracks.
The correction of functional posterior crossbite through Planas Direct Tracks has many characteristics that can become advantages. The aim of this study was to present a clinical case showing how to use this procedure for early correction with resources available through public health services. The patient, a 4-year-old girl, arrived to receive treatment due to a functional unilateral crossbite. When the mandible was moved to the centric position, it was observed that the teeth had occlusal trauma. An occlusal adjustment was performed. The adjustment was not sufficient to promote functional equilibrium; thus, Planas Direct Tracks were made, resulting in functional equilibrium and correction of the malocclusion. As shown in the case report, the Planas Direct Tracks were effective for the correction of the posterior crossbite. If malocclusion is considered a public health problem, implementation of low-cost and easy-to-execute techniques is needed.


To proliferate or to die: role of Id3 in cell cycle progression and survival of neural crest progenitors.
The neural crest is a unique population of mitotically active, multipotent progenitors that arise at the vertebrate neural plate border. Here, we show that the helix-loop-helix transcriptional regulator Id3 has a novel role in cell cycle progression and survival of neural crest progenitors in Xenopus. Id3 is localized at the neural plate border during gastrulation and neurulation, overlapping the domain of neural crest induction. Morpholino oligonucleotide-mediated depletion of Id3 results in the absence of neural crest precursors and a resultant loss of neural crest derivatives. This appears to be mediated by cell cycle inhibition followed by cell death of the neural crest progenitor pool, rather than a cell fate switch. Conversely, overexpression of Id3 increases cell proliferation and results in expansion of the neural crest domain. Our data suggest that Id3 functions by a novel mechanism, independent of cell fate determination, to mediate the decision of neural crest precursors to proliferate or die.


Attention with a mindful attitude attenuates subjective appetitive reactions and food intake following food-cue exposure.
Excessive energy intake that contributes to overweight and obesity is arguably driven by pleasure associated with the rewarding properties of energy-dense palatable foods. It is important to address influences of external food cues in food-abundant societies where people make over 200 food related decisions each day. This study experimentally examines protective effects of a mindful attention induction on appetitive measures, state craving and food intake following exposure to energy-dense foods. Forty females were randomly allocated to a standard food-cue exposure condition in which attention is brought to the hedonic properties of food or food-cue exposure following a mindful attention induction. Appetitive reactions were measured pre, post and 10 min after post-cue exposure, after which a plate of cookies was used as a surreptitious means of measuring food intake. Self-reported hunger remained unchanged and fullness significantly increased for the mindful attention group post-cue exposure whereas hunger significantly increased for the standard attention group and fullness remained unchanged. There was no significant between-group difference in state craving post-cue exposure and 10 min later. Significantly more cookies were eaten by the standard attention group 10 min post-cue exposure although no significant between-group differences in appetitive and craving measures were reported at that time. Our results point to a promising brief intervention strategy and highlights the importance of distinguishing mindful attention from attention. Results also demonstrate that mindful attention can influence food intake even when craving and hunger are experienced.