Aromatization of androgens by human adipose tissue in vitro.
Stromal cells prepared from adipose tissue of women were maintained in monolayer culture to study the regulation of aromatase activity by hormones. Aromatase activity was stimulated 20-100-fold by dexamethasone. Half-maximal stimulation of aromatase activity was attained at a dexamethasone concentration of 2.5 nM. The stimulatory effect of dexamethasone was apparent after a preincubation time of 4 h, and stimulation was maximal after 24 h of preincubation. The stimulatory effect of dexamethasone was observed only when fetal calf serum also was present in the culture medium. Of the various steroids tested, dexamethasone was the most potent in stimulating aromatase activity. Cytosolic fractions prepared from stromal cells that had been maintained in monolayer culture were found to contain a homogeneous population of sites that specifically bound [3H]-dexamethasone with relatively high affinity (Kd = 2.9 nM) and low capacity (38 fmol/mg of protein). The stimulatory effect of dexamethasone on aromatase activity was prevented by simultaneous incubation with cortisol 21-mesylate (0.1-10 microM), a compound known to block the binding of glucocorticosteroids to cytoplasmic receptors. These findings are suggestive that glucocorticosteroids act to increase aromatase activity in stromal cells by binding to a glucocorticosteroid receptor. This presumably results in the induction of synthesis of specific proteins, perhaps the form of cytochrome P-450 involved in aromatization, or else NADPH-cytochrome P-450 reductase. Aromatase activity was also stimulated by dibutyryl cyclic AMP ( (Bu)2cAMP). In contrast to the stimulation by dexamethasone, the effect of (Bu)2cAMP was diminished by 70% when fetal calf serum (15%) was present in the culture medium. Aromatase activity of cells incubated with (Bu)2cAMP continued to increase over an 8 day period, at which time, enzyme activity was increased by 200-fold. Aromatase activity of stromal cells maintained in the presence or absence of serum was unaffected after incubation for 5 days with oFSH, hCG, bPRL, hGH or soterenol. On the other hand, incubation of cells for 5 days with cholera toxin (10 micrograms/ml) or 1-methyl-3-isobutylxanthine (0.1 mM) resulted in a marked increase in aromatase activity; the effect of these agents was diminished greatly when serum was present in the culture medium. The findings that serum inhibited the stimulation of aromatase activity by (Bu)2cAMP and that the time courses of aromatase induction by (Bu)2cAMP and dexamethasone were markedly different are suggestive that different molecular events mediate aromatase induction by glucocorticosteroids and cyclic AMP.


Effects of prostaglandin E1 infusion in the pre-operative management of critical congenital heart disease.
Prostaglandin E1 (PGE1) was administered to 27 infants in whom pulmonary or systemic blood flow was entirely or significantly dependent upon the patency of the ductus arteriosus. In 12 patients with pulmonary atresia or severe pulmonary stenosis, PGE1 infusion was followed by an improvement in hypoxemia and acidemia (group I). In 2 patients with left ventricular outflow-tract obstruction, PGE1 infusion was followed by an improvement in arterial blood pressure, peripheral perfusion and urine output (group II). In 5 patients with d-transposition of the great arteries and intact ventricular septum who had persistent severe hypoxemia after creation of an interatrial communication, PGE1 infusion improved the arterial oxygenation with dilatation of the ductus arteriosus (group III). Seven patients (3 of group I, 2 of group II and 2 of group III) failed to respond to PGE1. There were no fatal side effects. It is concluded that PGE1 therapy is highly effective in stabilizing pre-operative conditions of infants with ductus-dependent congenital heart disease.


Simple, yet powerful methodologies for conformational sampling of proteins.
Several biological functions, such as molecular recognition, enzyme catalysis, signal transduction, allosteric regulation, and protein folding, are strongly related to conformational transitions of proteins. These conformational transitions are generally induced as slow dynamics upon collective motions, including biologically relevant large-amplitude fluctuations of proteins. Although molecular dynamics (MD) simulation has become a powerful tool for extracting conformational transitions of proteins, it might still be difficult to reach time scales of the biological functions because the accessible time scales of MD simulations are far from biological time scales, even if straightforward conventional MD (CMD) simulations using massively parallel computers are employed. Thus, it is desirable to develop efficient methods to achieve canonical ensembles with low computational costs. From this perspective, we review several enhanced conformational sampling techniques of biomolecules developed by us. In our methods, multiple independent short-time MD simulations are employed instead of single straightforward long-time CMD simulations. Our basic strategy is as follows: (i) selection of initial seeds (initial structures) for the conformational sampling in restarting MD simulations. Here, the seeds should be selected as candidates with high potential to transit. (ii) Resampling from the selected seeds by initializing velocities in restarting short-time MD simulations. A cycle of these simple protocols might drastically promote the conformational transitions of biomolecules. (iii) Once reactive trajectories extracted from the cycles of short-time MD simulations are obtained, a free energy profile is evaluated by means of umbrella sampling (US) techniques with the weighted histogram analysis method (WHAM) as a post-processing technique. For the selection of the initial seeds, we proposed four different choices: (1) Parallel CaScade molecular dynamics (PaCS-MD), (2) Fluctuation Flooding Method (FFM), (3) Outlier FLOODing (OFLOOD) method, and (4) TaBoo SeArch (TBSA) method. We demonstrate applications of our methods to several biological systems, such as domain motions of proteins with large-amplitude fluctuations, conformational transitions upon ligand binding, and protein folding/refolding to native structures of proteins. Finally, we show the conformational sampling efficiencies of our methods compared with those by CMD simulations and other previously developed enhanced conformational sampling methods.


[The teaching of nutrition at medical schools:current situation].
Although doctors, students and teachers all coincide in acknowledging the need for an appropriate knowledge of human nutrition in order to practice medicine, the inclusion of this discipline in the undergraduate curriculum has been rather scant. Until a few years ago, there were only two medical schools in Spain with a compulsory course in nutrition as part of the syllabus. An attempt will be made to assess the current status of the teaching of nutrition within the syllabuses for a degree in Medicine and Surgery at Spanish universities. The syllabuses of 27 Faculties of Medicine in Spain have been consulted to find out: 1) if there are specific subjects on nutrition, feeding and/or dietetics, 2) if these subjects are compulsory or optional, and 3) if there are specific issues from this area included within other subjects. The results of the query show that 7 schools (approximately 25% of the total) teach a compulsory subject on nutrition or dietetics whereas 17 have at least one optional subject in this field, particularly during the second or specialization stage, with a mean content worth 4.5 credits. Furthermore, 3 of the schools offer both compulsory and optional courses simultaneously. Within the course material of the core subjects included in the second stage of the degree, the subjects of Endocrinology, Metabolism and Paediatrics seem to be the ones most commonly including clinical nutrition topics, generally with a very small relative weight. The results obtained show an increasing interest in the creation of distinctive subjects for nutrition and/or dietetics. Nonetheless, this is still mainly an optional subject and the integration of human nutrition into the syllabus for core subjects seems insufficient for its to be considered a good alternative.


[Transmission of infectious oral diseases from mother to child].
The relation between oral microbes of mother and child encompasses several aspects. It has been shown that a bad periodontal condition is a risk factor for low birth weight. The bacterium that is most specifically associated with dental caries, Streptococcus mutans, can be transmitted between mother and child; it has been suggested that the age of colonization with this bacterium is a risk factor for caries in the child. Also periodontal bacteria can be transmitted between mother and child. It remains to be elucidated whether this is an additional risk factor for the development of periodontal disease.


Interaction between the G1057D variant of IRS-2 and overweight in the pathogenesis of type 2 diabetes.
The insulin receptor substrate-2 (IRS-2) is a major insulin signalling molecule. IRS-2 inactivation in mice induces a form of diabetes characterized by peripheral insulin resistance and reduced beta cell mass. We tested the hypothesis that a common non-conservative amino acid substitution of IRS-2 (G1057D) might interact with overweight in the pathogenesis of type 2 diabetes. The variant was genotyped in 193 Italian patients with type 2 diabetes and 206 control subjects. In the absence of overweight, the risk of type 2 diabetes decreased according to the dosage of the D1057 allele (odds ratio for GD genotype 0.46 [95% CI 0.25-0.86]; DD genotype 0.18 [0.04-0.68]; P for trend = 0.0012). Conversely, the interaction between overweight and genotype increased the risk of type 2 diabetes according to the dosage of the D1057 allele (odds ratio for GD genotype 2.50 [1.11-5.65]; DD genotype 5.74 [1.11-29. 78]; P for trend = 0.0047). Among controls, fasting C-peptide levels, after adjustment for plasma glucose, were inversely related to the dosage of the D1057 allele (P = 0.020). This finding suggested that carriers of the D1057 allele may have higher insulin sensitivity and supported the protective effect of this allele. Conversely, among overweight patients there was a parallel increase in fasting plasma glucose (P for trend = 0.037) and fasting C-peptide according to the dosage of the D1057 allele, suggesting that higher insulin resistance and relative beta cell failure contributed to the increased risk of type 2 diabetes in overweight carriers of this allele. These data provide evidence for a strong association between type 2 diabetes and the G1057D common genetic variant of IRS-2, which appears to be protective against type 2 diabetes in a codominant fashion. Overweight appears to modify the effect of this polymorphism toward a higher risk of type 2 diabetes. Carriers of this polymorphism may represent an elective target for prevention of type 2 diabetes through preventing or treating excessive weight.


Adaptive leadership framework for chronic illness: framing a research agenda for transforming care delivery.
We propose the Adaptive Leadership Framework for Chronic Illness as a novel framework for conceptualizing, studying, and providing care. This framework is an application of the Adaptive Leadership Framework developed by Heifetz and colleagues for business. Our framework views health care as a complex adaptive system and addresses the intersection at which people with chronic illness interface with the care system. We shift focus from symptoms to symptoms and the challenges they pose for patients/families. We describe how providers and patients/families might collaborate to create shared meaning of symptoms and challenges to coproduce appropriate approaches to care.


Differences in Memory, Perceptions, and Preferences of Multimedia Consumer Medication Information: Experimental Performance and Self-Report Study.
Electronic health resources are becoming prevalent. However, consumer medication information (CMI) is still predominantly text based. Incorporating multimedia into CMI (eg, images, narration) may improve consumers' memory of the information as well as their perceptions and preferences of these materials. This study examined whether adding images and narration to CMI impacted patients' (1) memory, (2) perceptions of comprehensibility, utility, or design quality, and (3) overall preferences. We presented 36 participants with CMI in 3 formats: (1) text, (2) text + images, and (3) narration + images, and subsequently asked them to recall information. After seeing all 3 CMI formats, participants rated the formats in terms of comprehensibility, utility, and design quality, and ranked them from most to least favorite. Interestingly, no significant differences in memory were observed (F2,70=0.1, P=0.901). Thus, this study did not find evidence to support multimedia or modality principles in the context of CMI. Despite the absence of effects on memory, the CMI format significantly impacted perceptions of the materials. Specifically, participants rated the text + images format highest in terms of comprehensibility (χ22=26.5, P<.001) and design quality (χ22=35.69, P<.001). Although the omnibus test suggested a difference in utility ratings as well (χ22=8.21, P=.016), no significant differences were found after correcting for multiple comparisons. Consistent with perception findings, the preference ranks yielded a significant difference (χ22=26.00, P<.001), whereby participants preferred the text + images format overall. Indeed, 75% (27/36) of participants chose the text + images format as their most favorite. Thus, although there were no objective memory differences between the formats, we observed subjective differences in comprehensibility, design quality, and overall preferences. This study revealed that although multimedia did not appear to influence memory of CMI, it did impact participants' opinions about the materials. The lack of observed differences in memory may have been due to ceiling effects, memory rather than understanding as an index of learning, the fragmented nature of the information in CMI itself, or the size or characteristics of the sample (ie, young, educated subjects with adequate health literacy skills). The differences in the subjective (ie, perceptions and preferences) and objective (ie, memory) results highlight the value of using both types of measures. Moreover, findings from this study could be used to inform future research on how CMI could be designed to better suit the preferences of consumers and potentially increase the likelihood that CMI is used. Additional research is warranted to explore whether multimedia impacts memory of CMI under different conditions (eg, older participants, subjects with lower levels of health literacy, more difficult stimuli, or extended time for decay).


A monoclonal antibody against activated protein C allows rapid detection of activated protein C in plasma and reveals a calcium ion dependent epitope involved in factor Va inactivation.
Activated protein C (APC) serves as an 'on demand' anticoagulant. Defects in the APC anticoagulant pathway are underlying risk factors for the development of venous and arterial thrombosis. APC has recently been shown to significantly reduce mortality in patients with severe sepsis, presumably by virtue of its ability to down-regulate coagulation as well as inflammation. Our objective was to develop an assay that, for the first time, permits rapid detection of plasma APC. This assay will expedite studies of APC in a variety of vascular disease states including sepsis, severe atherosclerosis, diabetes, and vasculitis. By generating a highly APC-specific monoclonal antibody (HAPC 1555), we have developed an assay that, for the first time, allows rapid detection of plasma APC. The Kd measured for the interaction between APC and HAPC 1555 based on BIAcore studies and binding to immobilized HAPC on microtiter plates is 6.2 +/- 0.9 and 8.8 +/- 1.0 nmol L(-1), respectively. The interaction between HAPC 1555 and APC is Ca2+-dependent, with a Ca2+ concentration of 313 +/- 48 micro mol L(-1) required for half maximal binding. HAPC 1555 interferes with APC-mediated inactivation of factor (F)Va in the presence and absence of phospholipids, suggesting that HAPC 1555 binds to the FVa binding domain of APC. When HAPC 1555 was used in an APC enzyme capture assay, therapeutic APC levels could be measured in 1.5 h, and physiologic levels of APC could be detected between 3 and 19 h. APC levels were also shown to vary markedly in patients with severe sepsis. The rapidity of our APC assay makes APC detection in patients practical clinically. This assay will expedite studies of APC in a variety of vascular disease states including sepsis, severe atherosclerosis, diabetes, and vasculitis.


Historic Museum Samples Provide Evidence for a Recent Replacement of Wolbachia Types in European Drosophila melanogaster.
Wolbachia is one of the most common bacterial endosymbionts, which is frequently found in numerous arthropods and nematode taxa. Wolbachia infections can have a strong influence on the evolutionary dynamics of their hosts since these bacteria are reproductive manipulators that affect the fitness and life history of their host species for their own benefit. Host-symbiont interactions with Wolbachia are perhaps best studied in the model organism Drosophila melanogaster, which is naturally infected with at least 5 different variants among which wMel and wMelCS are the most frequent ones. Comparisons of infection types between natural flies and long-term lab stocks have previously indicated that wMelCS represents the ancestral type, which was only very recently replaced by the nowadays dominant wMel in most natural populations. In this study, we took advantage of recently sequenced museum specimens of D. melanogaster that have been collected 90 to 200 yr ago in Northern Europe to test this hypothesis. Our comparison to contemporary Wolbachia samples provides compelling support for the replacement hypothesis. Our analyses show that sequencing data from historic museum specimens and their bycatch are an emerging and unprecedented resource to address fundamental questions about evolutionary dynamics in host-symbiont interactions. However, we also identified contamination with DNA from crickets that resulted in co-contamination with cricket-specific Wolbachia in several samples. These results underpin the need for rigorous quality assessments of museomic data sets to account for contamination as a source of error that may strongly influence biological interpretations if it remains undetected.


Does early surgical decompression in cauda equina syndrome improve bladder outcome?
We analyzed retrospectively whether early surgery for cauda equina syndrome (CES) within 24, 48, or 72 hours of onset of autonomic symptoms made any difference to bladder function at initial outpatient follow-up. CES potentially causes loss of autonomic control including bladder dysfunction, resulting in significant disability. There is significant debate regarding appropriate timing of surgery. We conducted a retrospective cohort study of 200 patients between 2000 and 2011 who underwent decompressive surgery for CES at a regional neurosurgical center. Data collected were from clinical admission and at initial follow-up. Presentation was categorized into CES with retention (CESR) and incomplete CES (CESI) and duration of autonomic symptoms before surgical intervention. A total of 200 patients had complete clinical records; 61 cases with CESR and 139 cases with CESI. Average initial follow-up time was 96 days. For the 36 cases with CESI less than 24 hours, normal bladder function was seen at follow-up in all patients except 4 (11.1%), but with 103 cases with CESI more than 24 hours, 48 (46.6%) had bladder dysfunction (Pearson χP = 0.000). For the 64 cases with CESI less than 48 hours, normal bladder function was seen at follow-up in all except 10 (15.6%), but with 75 cases with CESI more than 48 hours, 42 (56%) had bladder dysfunction (Pearson χP = 0.000). For the 35 patients with CESR, operating within 24, 48, or 72 hours made no obvious difference to bladder outcome. Data were also reanalyzed changing the dataset groups to CESI less than 24 hours, 24 to 48 hours, and more than 48 hours to calculate odds ratios regarding normal bladder outcome. We identified that decompressive surgery within 24 hours of onset of autonomic symptoms in CESI reduces bladder dysfunction at initial follow-up, but no statistically significant difference in outcome was observed in CESR regarding timing of operation. 3.


A Genetic Screen for Saccharomyces cerevisiae Mutants That Fail to Enter Quiescence.
Budding yeast begin the transition to quiescence by prolonging G1 and accumulating limited nutrients. They undergo asymmetric cell divisions, slow cellular expansion, acquire significant stress tolerance and construct elaborate cell walls. These morphologic changes give rise to quiescent (Q) cells, which can be distinguished from three other cell types in a stationary phase culture by flow cytometry. We have used flow cytometry to screen for genes that are required to obtain the quiescent cell fraction. We find that cell wall integrity is critical and these genes may help define quiescence-specific features of the cell wall. Genes required to evade the host innate immune response are common. These may be new targets for antifungal drugs. Acquired thermotolerance is also a common property, and we show that the stress-response transcription factors Msn2 and Msn4 promote quiescence. Many other pathways also contribute, including a subset of genes involved in autophagy, ubiquitin-mediated proteolysis, DNA replication, bud site selection, and cytokinesis.


Antigenic analysis of equine infectious anemia virus (EIAV) variants by using monoclonal antibodies: epitopes of glycoprotein gp90 of EIAV stimulate neutralizing antibodies.
Monoclonal antibodies produced against the prototype cell-adapted Wyoming strain of equine infectious anemia virus (EIAV), a lentivirus, were studied for reactivity with the homologous prototype and 16 heterologous isolates. Eighteen hybridomas producing monoclonal antibodies (MAbs) were isolated. Western blot (immunoblot) analyses indicated that 10 were specific for the major envelope glycoprotein (gp90) and 8 for the transmembrane glycoprotein (gp45). Four MAbs specific to epitopes of gp90 neutralized prototype EIAV infectivity. These neutralizing MAbs apparently reacted with variable regions of the envelope gp90, as evidenced by their unique reactivity with the panel of isolates, suggesting recognition of at least three different neutralization epitopes. The conformation of these epitopes appears to be continuous, as they resisted treatment with sodium dodecyl sulfate and reducing reagents. Monoclonal antibodies that reacted with conserved epitopes on gp90 or gp45 failed to neutralize EIAV. Our data also demonstrated that there was a large spectrum of possible EIAV serotypes and confirmed that antigenic variation occurs with high frequency in EIAV. Moreover, the data showed that variation is a rapid and random process, as no pattern of variant evolution was evident by comparison of 13 isolates from parallel infections. These results represent the first production of neutralizing MAbs specific for a lentivirus glycoprotein and document alterations in one or more neutralization epitopes of the major surface glycoprotein among sequential isolates of EIAV recovered during persistent infection.


Estimating the burden of mycetoma in Sudan for the period 1991-2018 using a model-based geostatistical approach.
Mycetoma is widespread in tropical and subtropical regions favouring arid areas with low humidity and a short rainy season. Sudan is one of the highly endemic countries for mycetoma. Estimating the population at risk and the number of cases is critical for delivering targeted and equitable prevention and treatment services. In this study, we have combined a large dataset of mycetoma cases recorded by the Mycetoma Research Centre (MRC) in Sudan over 28 years (1991-2018) with a collection of environmental and water and hygiene-related datasets in a geostatistical framework to produce estimates of the disease burden across the country. We developed geostatistical models to predict the number of cases of actinomycetoma and eumycetoma in areas considered environmentally suitable for the two mycetoma forms. Then used the raster dataset (gridded map) with the population estimates for 2020 to compute the potentially affected population since 1991. The geostatistical models confirmed this heterogeneous and distinct distribution of the estimated cases of eumycetoma and actinomycetoma across Sudan. For eumycetoma, these higher-risk areas were smaller and scattered across Al Jazirah, Khartoum, White Nile and Sennar states, while for actinomycetoma a higher risk for infection is shown across the rural districts of North and West Kurdufan. Nationally, we estimated 63,825 people (95%CI: 13,693 to 197,369) to have been suffering from mycetoma since 1991 in Sudan,51,541 people (95%CI: 9,893-166,073) with eumycetoma and 12,284 people (95%CI: 3,800-31,296) with actinomycetoma. In conclusion, the risk of mycetoma in Sudan is particularly high in certain restricted areas, but cases are ubiquitous across all states. Both prevention and treatment services are required to address the burden. Such work provides a guide for future control and prevention programs for mycetoma, highly endemic areas are clearly targeted, and resources are directed to areas with high demand.


P-(alkyl)-nucleoside 5'-hydrogenphosphonates as depot forms of antiviral nucleotide analogues.
P-(Alkyl)esters of AZT 5'-hydrogenphosphonate were synthesized and their stabilities in the phosphate buffer and human serum were evaluated. The esters bearing residues of primary and secondary alcohols were degraded to give AZT, whereas those containing tertiary alkyl groups yielded AZT 5'-hydrogenphosphonate. The corresponding derivatives of d2A and d4T showed the same properties.


3D retinal imaging and measurement using light field technology.
Light-field fundus photography has the potential to be a new milestone in ophthalmology. Up-to-date publications show only unsatisfactory image quality, preventing the use of depth measurements. We show that good image quality and, consequently, reliable depth measurements are possible, and we investigate the current challenges of this novel technology. We investigated whether light field (LF) imaging of the retina provides depth information, on which structures the depth is estimated, which illumination wavelength should be used, whether deeper layers are measurable, and what kinds of artifacts occur. The technical setup, a mydriatic fundus camera with an LF imager, and depth estimation were validated by an eye model and in vivo measurements of three healthy subjects and three subjects with suspected glaucoma. Comparisons between subjects and the corresponding optical coherence tomography (OCT) measurements were used for verification of the depth estimation. This LF setup allowed for three-dimensional one-shot imaging and depth estimation of the optic disc with green light. In addition, a linear relationship was found between the depth estimates of the OCT and those of the setup developed here. This result is supported by the eye model study. Deeper layers were not measurable. If image artifacts can be handled, LF technology has the potential to help diagnose and monitor glaucoma risk at an early stage through a rapid, cost-effective one-shot technology.


Monolayer film behavior of lipopolysaccharide from Pseudomonas aeruginosa at the air-water interface.
Lipopolysaccharide (LPS) is an essential biomacromolecule making up approximately 50% of the outer membrane of gram-negative bacteria. LPS chemistry facilitates cellular barrier and permeability functions and mediates interactions between the cell and its environment. To better understand the local interactions within LPS membranes, the monolayer film behavior of LPS extracted from Pseudomonas aeruginosa, an opportunistic pathogen of medical importance, was investigated by Langmuir film balance. LPS formed stable monolayers at the air-water interface and the measured lateral stresses and modulus (rigidity) of the LPS film in the compressed monolayer region were found to be appreciable. Scaling theories for two-dimensional (2D) polymer chain conformations were used to describe the pi-A profile, in particular, the high lateral stress region suggested that the polysaccharide segments reside at the 2D air-water interface. Although the addition of monovalent and divalent salts caused LPS molecules to adopt a compact conformation at the air-water interface, they did not appear to have any influence on the modulus (rigidity) of the LPS monolayer film under biologically relevant stressed conditions. With increasing divalent salt (CaCl2) content in the subphase, however, there is a progressive reduction of the LPS monolayer's collapse pressure, signifying that, at high concentrations, divalent salts weaken the ability of the membrane to withstand elevated stress. Finally, based on the measured viscoelastic response of the LPS films, we hypothesize that this property of LPS-rich outer membranes of bacteria permits the deformation of the membrane and may consequently protect bacteria from catastrophic structural failure when under mechanical-stress.


Agromyces allii sp. nov., isolated from the rhizosphere of Allium victorialis var. platyphyllum.
Two Gram-positive, non-motile, yellow-pigmented, slightly curved and rod-shaped bacterial strains, UMS-62(T) and UMS-101, were isolated from the rhizosphere of Allium victorialis var. platyphyllum, a variety of wild edible greens grown on Ulleung island, Korea. The taxonomic position of the strains was investigated by a polyphasic approach. Strains UMS-62(T) and UMS-101 grew optimally at 30 degrees C and at pH 6.5-7.5. The novel strains contained MK-11 and MK-12 as the predominant menaquinones and rhamnose, ribose and galactose as the major cell-wall sugars. The major cellular fatty acids (>10 % of the total fatty acids) were anteiso-C(15 : 0), iso-C(16 : 0) and anteiso-C(17 : 0). The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. The DNA G+C contents were 71.1-71.3 mol%. Phylogenetic trees based on 16S rRNA gene sequences showed that strains UMS-62(T) and UMS-101 belong to the genus Agromyces. Strains UMS-62(T) and UMS-101 showed a 16S rRNA gene sequence similarity value of 99.9 % and a mean DNA-DNA relatedness level of 91.1 %. Similarity values between the 16S rRNA gene sequences of the two novel strains and the type strains of recognized Agromyces species ranged from 95.2 to 99.1 %. The levels of DNA-DNA relatedness between the two novel strains and the type strains of five phylogenetically related Agromyces species were in the range of 13.4 to 54.2 %. On the basis of phenotypic properties, phylogenetic distinctiveness and genetic data, strain UMS-62(T) (=KCTC 19181(T)=JCM 13584(T)) and strain UMS-101 are classified as representing a novel species in the genus Agromyces, for which the name Agromyces allii sp. nov. is proposed.


Recovery of Polyphenols from Agri-Food By-Products: The Olive Oil and Winery Industries Cases.
The production of olive oil and wine are two of the main agri-food economic activities in Southern Europe. They generate large amounts of solid and liquid wastes (e.g., olive pomace, olive mill wastewater, grape pomace, grape stems, wine lees, and wine processing wastewater) that represent a major environmental problem. Consequently, the management of these residues has become a big challenge for these industries, since they are harmful to the environment but rich in bioactive compounds, such as polyphenols. In recent years, the recovery of phenolic compounds has been proposed as a smart strategy for the valorization of these by-products, from a circular economy perspective. This review aims to provide a comprehensive description of the state of the art of techniques available for the analysis, extraction, and purification of polyphenols from the olive mill and winery residues. Thus, the integration and implementation of these techniques could provide a sustainable solution to the olive oil and winery sectors.


Cancer screening behaviors among Korean-American women.
The goals of this study were to evaluate breast and cervical cancer screening tests and to examine the correlates of cancer screening behaviors. A cross-sectional face-to-face survey of 438 Korean-American women residing in Maryland was conducted. About 50% of women age 18 and older had had a Pap smear and 46.6% of these women age 40 and older had had a mammogram in the past 2 years. In multiple logistic regression analyses, the strongest correlate of screening behaviors was having a regular medical checkup. Age and acculturation were found to be important correlates of cancer screening tests: Women less than 50 years of age were more likely to have cancer screening tests than those 50 years and older. English language proficiency was associated with having a mammogram and the proportion of life spent in the United States was associated with having a Pap smear. Employment interacted with marital status for a Pap smear, with those married and unemployed being less likely to have a Pap smear than women who were both married and employed. For strategies to increase cancer-screening tests among Korean-American women, we need to aim at developing culturally appropriate educational programs about cancer for less acculturated and recent immigrants.


Rat basophilic leukemia cells stiffen when they secrete.
RBL cells provide a useful model of the IgE and antigen-dependent stimulus-secretion coupling of mast cells and basophils. We have measured cellular deformability to investigate the participation of cytoskeletal mechanical changes. Cross-linking cell-surface IgE-receptor complexes with multivalent ligands not only triggered secretion but also caused the cells to stiffen, i.e., to become more resistant to deformation. This mechanical response required receptor cross-linking, had a time course similar to that of secretion, and was reversed by DNP-L-lysine, a competitive inhibitor of antigen binding. Hence the same stimulus seems to elicit both stiffening and secretion. Cytochalasin D, which inhibits actin filament assembly, prevented or reversed stiffening, thereby implicating the cytoskeleton in the mechanical response. Increasing intracellular calcium ion concentration with the ionophore A23187 stiffened cells and stimulated secretion. Activation of protein kinase C with a phorbol ester also stiffened cells and enhanced both the stiffening and secretion caused by the ionophore. Yet cytochalasin D enhances secretion whereas activation of protein kinase c alone is insufficient for secretion. Therefore stiffening is neither necessary nor sufficient for secretion. These results characterize a cytoskeletal mechanical response triggered by the same receptor-dependent stimulus that elicits secretion and by second messengers that are thought to mediate between the receptor signal and secretion. The function of the mechanical response, however, remains to be determined.


Nursing students' self-efficacy in providing transcultural care.
The aim of any health care service is to provide optimal quality care to clients and families regardless of their ethnic group. As today's Australian society comprises a multicultural population that encompasses clients with different cultural norms and values, this study examined undergraduate nursing students' self-efficacy in providing transcultural nursing care. A sample of 196 nursing students enrolled in the first and fourth year of a pre-registration nursing program in a Western Australian University were invited to participate in a survey incorporating a transcultural self-efficacy tool (TSET) designed by Jeffery [Unpublished instrument copyrighted by author, 1994]. The findings revealed that fourth year students, exposed to increased theoretical information and clinical experience, had a more positive perception of their self-efficacy in providing transcultural nursing skills than the first year students. In addition, the study found that age, gender, country of birth, languages spoken at home and previous work experience did not influence the nursing students' perception of self-efficacy in performing transcultural care. The study supports the notion that educational preparation and relevant clinical experience is important in providing nursing students with the opportunity to develop self-efficacy in performing effective and efficient transcultural nursing in today's multicultural health care system. It is for this reason that educators need to focus on providing students with relevant theoretical information and ensure sufficient clinical exposure to support student learning in the undergraduate program.


Multivariate analyses of the profile stability of intelligence tests: high for IQs, low to very low for subtest analyses.
Profile stability involves the consistency of a set of scores over time. That is, does a profile of scores change on retesting and does this change affect clinical decisions? While psychologists routinely examine the reliability of individual scores, little research has examined the stability of a profile or set of scores. The first study described in this paper examined potential measures of profile stability using a simulation computer program. The results suggest that several measures show promise in this context, particularly Cattell's coefficient of pattern similarity (r(p)), salient variable similarity index (S), and the D(2) coefficient. In the second study, selected measures of profile stability were applied to Wechsler test-retest data. The results suggest that profiles composed of IQ and index scores demonstrate acceptable stability and can be usefully interpreted in clinical and research situations. However, subtest score profiles are inherently less stable and provide little useful clinical information.


Antimicrobial use and antimicrobial resistance in nosocomial pathogens at a tertiary care hospital in Pune.
Resistance to antimicrobial agents is emerging in wide variety of nosocomial and community acquired pathogens. Widespread and often inappropriate use of broad spectrum antimicrobial agents is recognized as a significant contributing factor to the development and spread of bacterial resistance. This study was conducted to gain insight into the prevalent antimicrobial prescribing practices, and antimicrobial resistance pattern in nosocomial pathogens at a tertiary care hospital in Pune, India. Series of one day cross sectional point prevalence surveys were carried out on four days between March and August 2014. All eligible in patients were included in the study. A structured data entry form was used to collect the data for each patient. Relevant samples were collected for microbiological examination from all the clinically identified hospital acquired infection cases. 41.73% of the eligible patients (95% CI: 39.52-43.97) had been prescribed at least one antimicrobial during their stay in the hospital. Beta-lactams (38%) were the most prescribed antimicrobials, followed by Protein synthesis inhibitors (24%). Majority of the organisms isolated from Hospital acquired infection (HAI cases) were found to be resistant to the commonly used antimicrobials viz: Cefotaxime, Ceftriaxone, Amikacin, Gentamicin and Monobactams. There is need to have regular antimicrobial susceptibility surveillance and dissemination of this information to the clinicians. In addition, emphasis on the rational use of antimicrobials, antimicrobial rotation and strict adherence to the standard treatment guidelines is very essential.


Effect of paper on the performance assay in the control of antibiotic sensitivity discs.
Antibiotic discs were prepared, using several several batches of papers meeting Food and Drug Administration specifications. The analysis of 1,152 zones of inhibition produced showed no performance differences among these batches. Other discs were prepared using papers of different grades. These produced large differences in performance. It is obvious, therefore, that the use of a specified disc paper is necessary for standardizing the performances of the products of various manufacturers and that reproducible results can be attained with the grade of paper specified.


Oxytocin and the oxytocin receptor underlie intrastrain, but not interstrain, social recognition.
We studied three lines of oxytocin (Oxt) and oxytocin receptor (Oxtr) knockout (KO) male mice [Oxt(-/-), total Oxtr(-/-) and partial forebrain Oxtr (Oxtr(FB/FB))] with established deficits in social recognition to further refine our understanding of their deficits with regard to stimulus female's strain. We used a modified social discrimination paradigm in which subjects are singly housed only for the duration of the test. Additionally, stimulus females are singly housed throughout testing and are presented within corrals for rapid comparison of investigation by subject males. Wild-type (WT) males from all three lines discriminated between familiar and novel females of three different strains (C57BL/6, BALB/c and Swiss-Webster). No KO males discriminated between familiar and novel BALB/c or C57BL/6 females. Male Oxt(-/-) and Oxtr(-/-) mice, but not Oxtr(FB/FB) mice, discriminated between familiar and novel Swiss-Webster females. As this might indicate a global deficit in individual recognition for Oxtr(FB/FB) males, we examined their ability to discriminate between females from different strains and compared performance with Oxtr(-/-) males. WT and KO males from both lines were able to distinguish between familiar and novel females from different strains, indicating the social recognition deficit is not universal. Instead, we hypothesize that the Oxtr is involved in 'fine' intrastrain recognition, but is less important in 'broad' interstrain recognition. We also present the novel finding of decreased investigation across tests, which is likely an artifact of repeated testing and not because of stimulus female's strain or age of subject males.


Accumulation of the sigma-1 receptor is common to neuronal nuclear inclusions in various neurodegenerative diseases.
The sigma-1 receptor (SIGMAR1) is now known to be one of the endoplasmic reticulum (ER) chaperones, which participate in the degradation of misfolded proteins in cells via the ER-related degradation machinery linked to the ubiquitin-proteasome pathway. Mutations of the SIGMAR1 gene are implicated in the pathogenesis of familial frontotemporal lobar degeneration and motor neuron disease. Involvement of ER dysfunction in the formation of inclusion bodies in various neurodegenerative diseases has also become evident. We performed immunohistochemical staining to clarify the localization of SIGMAR1 in the brains of patients with neurodegenerative disorders, including trans-activation response DNA protein 43 (TDP-43) proteinopathy, tauopathy, α-synucleinopathy, polyglutamine disease and intranuclear inclusion body disease (INIBD). Double-immunocytofluorescence and Western blot analyses of cultured cells were also performed to investigate the role of SIGMAR1 using a specific exportin 1 inhibitor, leptomycin B and an ER stress inducer, thapsigargin. SIGMAR1 was consistently shown to be co-localized with neuronal nuclear inclusions in TDP-43 proteinopathy, five polyglutamine diseases and INIBD, as well as in intranuclear Marinesco bodies in aged normal controls. Cytoplasmic inclusions in neurons and glial cells were unreactive for SIGMAR1. In cultured cells, immunocytofluorescent study showed that leptomycin B and thapsigargin were shown to sequester SIGMAR1 within the nucleus, acting together with p62. This finding was also supported by immunoblot analysis. These results indicate that SIGMAR1 might shuttle between the nucleus and the cytoplasm. Neurodegenerative diseases characterized by neuronal nuclear inclusions might utilize the ER-related degradation machinery as a common pathway for the degradation of aberrant proteins.


ALS drug development guidances and trial guidelines: Consensus and opportunities for alignment.
The US Food and Drug Administration (FDA) developed a draft guidance for drug development in amyotrophic lateral sclerosis (ALS) that was issued in February 2018. The FDA draft guidance considered the recommendations developed by the ALS community that incorporated the views of a large group of clinical investigators, industry representatives, advocacy groups, patients, and caregivers. This external input from the ALS community reviewed the current state of clinical research in ALS, made suggestions over a wide range of drug development topics, and served as an educational tool to provide the agency with additional inputs about ALS, the state of the science, and the community's views on key topics. In parallel to this effort, there was an independent effort to revise and update the ALS Clinical Trial Guidelines. We discuss the areas of agreement of these 3 documents and the areas that provide opportunities to improve the efficiency of drug development in ALS. It is likely that further research into biomarkers, efficacy endpoints, and predictive algorithms will provide greater alignment among community stakeholders and increase clarity on drug development efforts going forward. Continued patient engagement and inclusion of patient experience data in every aspect of the drug development process will further facilitate the approval of new treatments.


Lifetime risk of stroke in Japan.
Lifetime risk (LTR) is an epidemiologic measure that expresses the probability of disease in the remaining lifetime for an index age. The LTR for stroke has not been reported for the Japanese population. We included all participants from the Suita Study who were cardiovascular disease-free at baseline. Age (in years) was used as the time scale. Age-specific stroke incidence and all-cause mortality were calculated with the person-year method, and we estimated the sex- and index age-specific LTRs of first-ever stroke and its subtypes, taking into account the competing risk of death. We followed up 5498 participants from 1989 to 2005 for a total of 67 475 person-years. At age 55 years, the LTR for stroke, after accounting for competing risks of death, was 18.3% for men and 19.6% for women. The LTR for cerebral infarction was 14.6% for men and 15.5% for women, and the LTR for intracerebral hemorrhage was 2.4% for men and 1.4% for women at the index age of 55 years. The LTR for stroke remained similar across other index ages of 45, 55, and 65 years. The observed probabilities illustrate that approximately 1 in 5 men and women of middle age will experience stroke in their remaining lifetime. This easy understandable information can be used as an important index to assist in public health education and planning.


[Activities of glutathione enzymes in liver biopsy specimens in chronic lesions of hepatocytes].
The nonenzymatic conjugation of metabolites is decreased in chronic diseases of the liver, which is caused by decreased concentration of glutathione. The activities of glutathione enzymes are increased, this indicating the development of compensatory processes of mobilization of the second phase of detoxication, that is, increased conjugation under conditions of suppression of the cytochrome P-450 system. Measurement of liver glutathione transferase is a highly informative test for assessing the activity of the pathological process, particularly important in patients with chronic active hepatitis and cirrhosis of the liver.


Source document verification in the Mucopolysaccharidosis Type I Registry.
PURPOSE: The Mucopolysaccharidosis Type I (MPS I) Registry is an international observational database that tracks the natural history and the outcomes of patients with MPS I. The Registry was a regulatory requirement following the approval of laronidase enzyme replacement therapy for MPS I in 2003. All data are collected voluntarily after informed consent from the patient or family. Data are checked through queries, monthly reviews, and electronic audits to identify missing, inconsistent, or invalid data. This analysis sought to determine overall data accuracy in the Registry through source document verification (SDV). METHODS: Two phases of SDV were performed. In each phase, Registry data were compared against source documents at sites in Europe, Latin America, and North America. Three patients were randomly selected for SDV at each of the selected sites among all patients enrolled ≥18 months and ever receiving laronidase. Key parameters central to MPS I and its treatment were examined from the baseline and the last available assessments. RESULTS: Results indicate an overall source-to-database error rate in the MPS I Registry of 2.7% (47 discrepancies out of 1715 items; 95% confidence interval [2.2%, 3.5%]) in Phase 1 and 3.7% (64 discrepancies out of 1732 items; 95% confidence interval [2.9%, 4.7%]) in Phase 2. No systematic errors were found. CONCLUSIONS: The overall error rates in both phases of SDV demonstrate acceptable data accuracy in the MPS I Registry within the data fields that were assessed. Copyright © 2011 John Wiley & Sons, Ltd.


Circular dichroism of polynucleotides: Interactions of NiCl2 with poly(dA-dT).poly(dA-dT) and poly(dG-dC).poly(dG-dC) in a water-in-oil microemulsion.
The thermal behavior of the synthetic, high molecular weight, double stranded polynucleotides poly(dA-dT).poly(dA-dT) [polyAT] and poly(dG-dC).poly(dG-dC) [polyGC] solubilized in the aqueous core of the quaternary water-in-oil cationic microemulsion CTAB|n-pentanol|n-hexane|water in the presence of increasing amounts of NiCl(2) at several constant ionic strength values (NaCl) has been studied by means of circular dichroism and electronic absorption spectroscopies. In the microemulsive medium, both polynucleotides show temperature-induced modifications that markedly vary with both Ni(II) concentration and ionic strength. An increase of temperature causes denaturation of the polyAT duplex at low nickel concentrations, while more complex CD spectral modifications are observed at higher nickel concentrations and ionic strengths. By contrast, thermal denaturation is never observed for polyGC. At low Ni(II) concentrations, the increase of temperature induces conformational transitions from B-DNA to Z-DNA form, or, more precisely, to left-handed helical structures. In some cases, at higher nickel concentrations, the CD spectra suggest the presence of Z'-type forms of the polynucleotide.


Effectiveness of quenchers to reduce radiolysis of (111)In- or (177)Lu-labelled methionine-containing regulatory peptides. Maintaining radiochemical purity as measured by HPLC.
An overview how to measure and to quantify radiolysis by the addition of quenchers and to maintain Radio-Chemical Purity (RCP) of vulnerable methionine-containing regulatory peptides is presented. High RCP was only achieved with a combination of quenchers. However, quantification of RCP is not standardized, and therefore comparison of radiolabelling and RCP of regulatory peptides between different HPLC-systems and between laboratories is cumbersome. Therefore we suggest a set of standardized requirements to quantify RCP by HPLC for radiolabelled DTPA- or DOTA-peptides. Moreover, a dosimetry model was developed to calculate the doses in the reaction vials during radiolabelling and storage of the radiopeptides, and to predict RCP in the presence and absence of quenchers. RCP was measured by HPLC, and a relation between radiation dose and radiolysis of RCP was established. The here described quenchers are tested individually as ƒ(concentration) to investigate efficacy to reduce radiolysis of radiolabelled methionine-containing regulatory peptides.


Unilateral pallidotomy as a potential rescue therapy for cervical dystonia after unsatisfactory selective peripheral denervation.
Selective peripheral denervation (SPD) is a widely accepted surgery for medically refractory cervical dystonia (CD), but when SPD has failed, the available approaches are limited. The authors investigated the results from a cohort of CD patients treated with unilateral pallidotomy after unsatisfactory SPD. The authors retrospectively analyzed patients with primary CD who underwent unilateral pallidotomy after SPD between April 2007 and August 2019. The Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) was used to evaluate symptom severity before surgery, 7 days postsurgery, 3 months postsurgery, and at the last follow-up. TWSTRS subscores for disability and pain and the 24-item Craniocervical Dystonia Questionnaire (CDQ-24) were used to assess quality of life. At a mean final follow-up of 5 years, TWSTRS severity subscores and total scores were significantly improved (n = 12, mean improvement 57.3% and 62.3%, respectively, p = 0.0022 and p = 0.0022), and 8 of 12 patients (66.7%) were characterized as responders (improvement ≥ 25%). Patients with rotation symptoms before pallidotomy showed greater improvement in TWSTRS severity subscores than those who did not (p = 0.049). The most common adverse event was mild upper-limb weakness (n = 3). Patients' quality of life was also improved. Unilateral pallidotomy seems to offer an effective and safe option for patients with CD who have otherwise experienced limited benefits from SPD.


A 3D porous FeP/rGO modulated separator as a dual-function polysulfide barrier for high-performance lithium sulfur batteries.
Lithium-sulfur batteries (LSBs) have gained considerable attention for their desirable energy densities, high theoretical capacities, low cost and environmentally friendly properties. However, the shuttle effect of polysulfides seriously hinders their future practical applications. Herein, a dual-function cathode structure, consisting of 3D porous FeP/rGO microspheres supported on both aluminum foil and a commercial separator, exhibits excellent performance by providing strong adsorption with respect to Li2Sx (x = 1, 2, 4, 6 and 8) and S8. In this rational design, the iron phosphide (FeP) nanoparticles act as a catalyst to accelerate polysulfide conversion and as the designated sites for adsorption. The 3D rGO porous conductive network can provide enough space for sulfur loading and to physically adsorb the polysulfides. More importantly, density functional theory (DFT) calculations also verified the strong interactions (with adsorption energy values of -4.21 to -1.97 eV) between the FeP(111) surface and the sulfur species. The electrochemical results show that the cell using the dual-function cathode structure delivers a capacity of 925.7 mA h g-1, with capacity degradation of 0.05% per cycle after 500 cycles, at a current density of 0.5C. It is also worth mentioning that the cell with sulfur loading of ∼2.2 mg cm-2 maintained a high capacity of 483 mA h g-1 at 0.5C after 500 cycles. In summary, the above results demonstrate the promising application of the dual-function cathode structure for high-performance LSBs.


Mortality from flash floods: a review of national weather service reports, 1969-81.
Of all weather-related disasters that occur in the United States, floods are the main cause of death, and most flood-related deaths are attributed to flash floods. Whenever a weather-related disaster involves 30 or more deaths or more than $100 million in property damage, the National Weather Service (NWS) forms a survey team to investigate the disaster and write a report of findings. All NWS survey reports on flash floods issued during 1969-81 were reviewed to determine the mortality resulting from such floods, the effect of warnings on mortality, and the circumstances contributing to death. A total of 1,185 deaths were associated with 32 flash floods, an average of 37 deaths per flash flood. The highest average number of deaths per event was associated with the four flash floods in which dams broke after heavy rains. Although there were 18 flash floods in 1977-81 and only 14 in 1969-76, the number of deaths was 2 1/2 times greater during the earlier period. More than twice as many deaths were associated with flash floods for which the survey team considered the warnings inadequate than with those with warnings considered adequate. Ninety-three percent of the deaths were due to drowning and 42 percent of these drownings were car related. The other drownings occurred in homes, at campsites, or when persons were crossing bridges and streams. The need for monitoring dams during periods of heavy rainfall is highlighted.


Reduction of hepatitis B seroprevalence in blood banking units by combined utilization of self-exclusion forms and clinical evaluation of blood donation candidates in Turkey.
In this retrospectively designed study, we evaluate the influence of the permanence and qualifications of health-care professionals on blood disposal rates due to hepatitis seropositivity. We observed a decrease of 44.2% in the number of blood units being rejected due to the donor's hepatitis B seropositivity in the second study period in which self-exclusion forms and where blood donation candidates were evaluated by a family physician. However, a similar decrease of the disposal rate due to hepatitis C seropositivity was not observed. This is especially important in countries which cannot afford many of the expensive modern laboratory tests. A comprehensive evaluation of self-exclusion forms and a brief examination prior to donation will greatly increase transfusion safety. Unfortunately, there are blood banks in Turkey which employ no physicians at all.


Lithium-Sodium Beta Alumina: First of a Family of Co-ionic Conductors?
Lithium-sodium beta alumina having a lithium/sodium ratio greater than about I appears to be the first generally useful lithium superionic conductor that has been reported. It exhibits strikingly nonlinear ion exchange properties and may presage the discovery of similar co-ionic interactions in other superionic conductors. The properties of lithium-sodium beta alumina are discussed in relation to current concepts of ionic interaction and distribution in the beta alumina conduction plane.


Sir Frederick Smith Memorial Lecture. A superb transport system--the circulation.
This paper discusses certain aspects of circulatory physiology, including foetal circulation and the role of the thorax, and gives parameters for total blood volume, cardiac output, stroke volume and the distribution of blood between the pulmonary and peripheral circulation within the horse. The reader is taken on a journey around the circulatory system, beginning in the left atrium, into which richly oxygenated blood runs from the pulmonary veins. From here the blood is squeezed via the mitral valve into the left ventricular pump, where it is forced under high pressure through the peripheral circulation, the arterioles and capillaries, to collect, deoxygenated and under much lower pressure, into the venules and veins. The pumping action of the muscles and the negative intrathoracic pressure help to draw the blood back up to the right atrium, itself exerting a suction effect on the great veins. The blood is then expelled from the right ventricle into the low resistance, high capacitance pulmonary circuit, where it picks up oxygen, releases carbon dioxide and returns to the left atrium.


Uromodulin upregulates TRPV5 by impairing caveolin-mediated endocytosis.
Uromodulin (UMOD) is synthesized in the thick ascending limb and secreted into urine as the most abundant protein. Association studies in humans suggest protective effects of UMOD against calcium-containing kidney stones. Mice carrying mutations of Umod found in human UMOD-associated kidney disease (UAKD) and Umod-deficient mice exhibit hypercalciuria. The mechanism for UMOD regulation of urinary Ca(2+) excretion is incompletely understood. We examined if UMOD regulates TRPV5 and TRPV6, channels critical for renal transcellular Ca(2+) reabsorption. Coexpression with UMOD increased whole-cell TRPV5 current density in HEK293 cells. In biotinylation studies, UMOD increased TRPV5 cell-surface abundance. Extracellular application of purified UMOD upregulated TRPV5 current density within physiological relevant concentration ranges. UMOD exerted a similar effect on TRPV6. TRPV5 undergoes constitutive caveolin-mediated endocytosis. UMOD had no effect on TRPV5 in a caveolin-1-deficient cell line. Expression of recombinant caveolin-1 in these cells restored the ability of UMOD to upregulate TRPV5. Secretion of UAKD-mutant UMOD was markedly reduced and coexpression of mutant UMOD with TRPV5 failed to increase its current. Immunofluorescent studies demonstrated lower TRPV5 expression in Umod(-/-) mice compared with wild-type. UMOD upregulates TRPV5 by acting from extracellular and by decreasing endocytosis of TRPV5. The stimulation of Ca(2+) reabsorption via TRPV5 by UMOD may contribute to protection against kidney-stone formation.


Mechanism of radiosensitization by the Chk1/2 inhibitor AZD7762 involves abrogation of the G2 checkpoint and inhibition of homologous recombinational DNA repair.
The median survival for patients with locally advanced pancreatic cancer treated with gemcitabine and radiation is approximately 1 year. To develop improved treatment, we have combined a Chk1/2-targeted agent, AZD7762, currently in phase I clinical trials, with gemcitabine and ionizing radiation in preclinical pancreatic tumor models. We found that in vitro AZD7762 alone or in combination with gemcitabine significantly sensitized MiaPaCa-2 cells to radiation. AZD7762 inhibited Chk1 autophosphorylation (S296 Chk1), stabilized Cdc25A, and increased ATR/ATM-mediated Chk1 phosphorylation (S345 Chk1). Radiosensitization by AZD7762 was associated with abrogation of the G(2) checkpoint as well as with inhibition of Rad51 focus formation, inhibition of homologous recombination repair, and persistent gamma-H2AX expression. AZD7762 was also a radiation sensitizer in multiple tumor xenograft models. In both MiaPaCa-2- and patient-derived xenografts, AZD7762 significantly prolonged the median time required for tumor volume doubling in response to gemcitabine and radiation. Together, our findings suggest that G(2) checkpoint abrogation and homologous recombination repair inhibition both contribute to sensitization by Chk1 inhibition. Furthermore, they support the clinical use of AZD7762 in combination with gemcitabine and radiation for patients with locally advanced pancreatic cancer.


Role of Porcine Aminopeptidase N and Sialic Acids in Porcine Coronavirus Infections in Primary Porcine Enterocytes.
Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) have been reported to use aminopeptidase N (APN) as a cellular receptor. Recently, the role of APN as a receptor for PEDV has been questioned. In our study, the role of APN in PEDV and TGEV infections was studied in primary porcine enterocytes. After seven days of cultivation, 89% of enterocytes presented microvilli and showed a two- to five-fold higher susceptibility to PEDV and TGEV. A significant increase of PEDV and TGEV infection was correlated with a higher expression of APN, which was indicative that APN plays an important role in porcine coronavirus infections. However, PEDV and TGEV infected both APN positive and negative enterocytes. PEDV and TGEV Miller showed a higher infectivity in APN positive cells than in APN negative cells. In contrast, TGEV Purdue replicated better in APN negative cells. These results show that an additional receptor exists, different from APN for porcine coronaviruses. Subsequently, treatment of enterocytes with neuraminidase (NA) had no effect on infection efficiency of TGEV, implying that terminal cellular sialic acids (SAs) are no receptor determinants for TGEV. Treatment of TGEV with NA significantly enhanced the infection which shows that TGEV is masked by SAs.


Glioplasticity in irritable bowel syndrome.
Growing evidence indicates a wide array of cellular remodeling in the mucosal microenvironment during irritable bowel syndrome (IBS), which possibly contributes to pathophysiology and symptom generation. Here, we investigated whether enteric glial cells (EGC) may be altered, and which factors/mechanisms lead to these changes. Colonic mucosal biopsies of IBS patients (13 IBS-Constipation [IBS-C]; 10 IBS-Diarrhea [IBS-D]; 11 IBS-Mixed [IBS-M]) and 24 healthy controls (HC) were analyzed. Expression of S100β and GFAP was measured. Cultured rat EGC were incubated with supernatants from mucosal biopsies, then proliferation and Ca2+ response to ATP were analyzed using flow cytometry and Ca2+ imaging. Histamine and histamine 1-receptor (H1R) involvement in the effects of supernatant upon EGC was analyzed. Compared to HC, the mucosal area immunoreactive for S100β was significantly reduced in biopsies of IBS patients, independently of the IBS subtype. IBS-C supernatants reduced EGC proliferation and IBS-D and IBS-M supernatants reduced Ca2+ response to ATP in EGC. EGC expressed H1R and the effects of supernatant upon Ca2+ response to ATP in EGC were blocked by pyrilamine and reproduced by histamine via H1R. IBS supernatants reduced mRNA expression of connexin-43. The S100β-stained area was negatively correlated with the frequency and intensity of pain and bloating. Changes in EGC occur in IBS, involving mucosal soluble factors. Histamine, via activation of H1R-dependent pathways, partly mediates altered Ca2+ response to ATP in EGC. These changes may contribute to the pathophysiology and the perception of pain and bloating in patients with IBS.


Mammalian Cdc7-Dbf4 protein kinase complex is essential for initiation of DNA replication.
The Cdc7-Dbf4 kinase is essential for regulating initiation of DNA replication in Saccharomyces cerevisiae. Previously, we identified a human Cdc7 homolog, HsCdc7. In this study, we report the identification of a human Dbf4 homolog, HsDbf4. We show that HsDbf4 binds to HsCdc7 and activates HsCdc7 kinase activity when HsDbf4 and HsCdc7 are coexpressed in insect and mammalian cells. HsDbf4 protein levels are regulated during the cell cycle with a pattern that matches that of HsCdc7 protein kinase activity. They are low in G(1), increase during G(1)-S, and remain high during S and G(2)-M. Purified baculovirus-expressed HsCdc7-HsDbf4 selectively phosphorylates the MCM2 subunit of the minichromosome maintenance (MCM) protein complex isolated by immunoprecipitation with MCM7 antibodies in vitro. Two-dimensional tryptic phosphopeptide-mapping analysis of in vivo (32)P-labeled MCM2 from HeLa cells reveals that several major tryptic phosphopeptides of MCM2 comigrate with those of MCM2 phosphorylated by HsCdc7-HsDbf4 in vitro, suggesting that MCM2 is a physiological HsCdc7-HsDbf4 substrate. Immunoneutralization of HsCdc7-HsDbf4 activity by microinjection of anti-HsCdc7 antibodies into HeLa cells blocks initiation of DNA replication. These results indicate that the HsCdc7-HsDbf4 kinase is directly involved in regulating the initiation of DNA replication by targeting MCM2 protein in mammalian cells.


Real-time image-content-based beamline control for smart 4D X-ray imaging.
Real-time processing of X-ray image data acquired at synchrotron radiation facilities allows for smart high-speed experiments. This includes workflows covering parameterized and image-based feedback-driven control up to the final storage of raw and processed data. Nevertheless, there is presently no system that supports an efficient construction of such experiment workflows in a scalable way. Thus, here an architecture based on a high-level control system that manages low-level data acquisition, data processing and device changes is described. This system is suitable for routine as well as prototypical experiments, and provides specialized building blocks to conduct four-dimensional in situ, in vivo and operando tomography and laminography.


Kap120 functions as a nuclear import receptor for ribosome assembly factor Rpf1 in yeast.
The nucleocytoplasmic exchange of macromolecules is mediated by receptors specialized in passage through the nuclear pore complex. The majority of these receptors belong to the importin beta protein family, which has 14 members in Saccharomyces cerevisiae. Nine importins carry various cargos from the cytoplasm into the nucleus, whereas four exportins mediate nuclear export. Kap120 is the only receptor whose transport cargo has not been found previously. Here, we characterize Kap120 as an importin for the ribosome maturation factor Rpf1, which was identified in a two-hybrid screen. Kap120 binds directly to Rpf1 in vitro and is released by Ran-GTP. At least three parallel import pathways exist for Rpf1, since nuclear import is defective in strains with the importins Kap120, Kap114, and Nmd5 deleted. Both kap120 and rpf1 mutants accumulate large ribosomal subunits in the nucleus. The nuclear accumulation of 60S ribosomal subunits in kap120 mutants is abolished upon RPF1 overexpression, indicating that Kap120 does not function in the actual ribosomal export step but rather in import of ribosome maturation factors.


TLR2/TLR4-dependent exaggerated cytokine production in hyperimmunoglobulinaemia D and periodic fever syndrome.
The autoinflammatory hyperimmunoglobulinaemia D and periodic fever syndrome (HIDS) is characterized by recurrent episodes of fever and inflammation. As part of the mevalonate kinase deficiency spectrum, it is caused by MVK mutations, resulting in decreased mevalonate kinase activity in the isoprenoid pathway. Although IL-1β is considered a major cytokine in its pathogenesis, IL-1 blockade is not successful in a proportion of patients. We aimed to further characterize the pro-inflammatory cytokine profile of HIDS. Peripheral blood mononuclear cells from HIDS patients and healthy donors were incubated with several stimuli. Cytokine concentrations were detected by ELISA. To analyse mRNA and protein expression, we performed quantitative RT-PCR and western blot, respectively. We observed significant differences in cytokine production when cells were incubated with ligands for Toll-like receptor 2 (TLR2), TLR4 and nucleotide-binding oligomerization domain-containing 2 (NOD2). The increased ratio between active and inactive caspase-1 protein in HIDS patients could explain why these cells are more easily triggered to secrete IL-1β. This is apparently not regulated at the transcriptional level, since expression levels of caspase-1 and IL-1β mRNA were similar in patients and controls. Both anakinra and tocilizumab treatment resulted in decreased inflammation, both ex vivo as well as in vivo. The increased cytokine secretion in HIDS is specific for TLR2, TLR4 and NOD2 ligation. Although IL-1β is important in the HIDS pathology, our data suggest it is a multicytokine disease. A more rigorous clinical trial is required to determine whether IL-6 receptor blockade may be considered in patients not responding to anakinra treatment.


Nonrandom chromosome abnormalities in acute leukemia and dysmyelopoietic syndromes in patients with previously treated malignant disease.
Cytogenetic studies were performed on 26 patients who developed acute nonlymphocytic leukemia (ANLL) or a dysmyelopoietic syndrome after treatment of a primary malignancy. Fifteen patients had radiotherapy and chemotherapy, seven had only chemotherapy, and four had only radiotherapy. The median times from diagnosis of the initial disease to the development of bone marrow dysfunction for these treatment groups were 50, 46, and 49 mo, respectively. Twenty-five patients had an abnormal karyotype in myeloid cells. Loss of part or all of no. 5 and/or no. 7 was noted in 23 of 25 patients with aneuploidy. Loss of no. 5 was noted only in patients who previously had malignant lymphoma, whereas loss of no. 7 was seen in these patients as well as in those who had other malignancies. Abnormalities of both nos. 5 and 7 occurred in 53% of the patients treated with combined therapy and in only 27% of patients treated with either modality alone. Although these changes are distinctly different from those noted in lymphomas, they are similar to those seen in 25% of aneuploid patients with ANLL de novo.


Wound-healing outcomes using standardized assessment and care in clinical practice.
Wound-healing outcomes applying standardized protocols have typically been measured within controlled clinical trials, not natural settings. Standardized protocols of wound care have been validated for clinical use, creating an opportunity to measure the resulting outcomes. Wound-healing outcomes were explored during clinical use of standardized validated protocols of care based on patient and wound assessments. This was a prospective multicenter study of wound-healing outcomes management in real-world clinical practice. Healing outcomes from March 26 to October 31, 2001, were recorded on patients in 3 long-term care facilities, 1 long-term acute care hospital, and 12 home care agencies for wounds selected by staff to receive care based on computer-generated validated wound care algorithms. After diagnosis, wound dimensions and status were assessed using a tool adapted from the Pressure Sore Status Toolfor use on all wounds. Wound, ostomy, and continence nursing professionals accessed consistent protocols of care, via telemedicine in home care or paper forms in long-term care. A physician entered assessments into a desktop computer in the wound clinic. Based on evidence that healing proceeds faster with fewer infections in environments without gauze, the protocols generally avoided gauze dressings. Most of the 767 wounds selected to receive the standardized-protocols of care were stage III-IV pressure ulcers (n = 373; mean healing time 62 days) or full-thickness venous ulcers (n = 124; mean healing time 57 days). Partial-thickness wounds healed faster than same-etiology full-thickness wounds. These results provide benchmarks for natural-setting healing outcomes and help to define and address wound care challenges. Outcomes primarily using nongauze protocols of care matched or surpassed best previously published results on similar wounds using gauze-based protocols of care, including protocols applying gauze impregnated with growth factors or other agents.


Natural ligand of mouse CD1d1: cellular glycosylphosphatidylinositol.
Mouse CD1d1, a member of the CD1 family of evolutionarily conserved major histocompatibility antigen-like molecules, controls the differentiation and function of a T lymphocyte subset, NK1+ natural T cells, proposed to regulate immune responses. The CD1d1 crystal structure revealed a large hydrophobic binding site occupied by a ligand of unknown chemical nature. Mass spectrometry and metabolic radiolabeling were used to identify cellular glycosylphosphatidylinositol as a major natural ligand of CD1d1. CD1d1 bound glycosylphosphatidylinositol through its phosphatidylinositol aspect with high affinity. Glycosylphosphatidylinositol or another glycolipid could be a candidate natural ligand for CD1d1-restricted T cells.


Discrimination of different feed additives and poly-herbal formulations based on their untargeted phytochemical profiles.
Feed additives represents a valid tool in animal nutrition to improve animal performance and livestock productivity under a sustainable perspective; however, there is a paucity of information about their comprehensive metabolomic and bioactive profiles. In this study, we tested the ability of an untargeted metabolomics approach to discriminate nine commercial feed additives and unique blends of botanical extracts used in both ruminant and non-ruminant nutrition, according to their phytochemical profiles and different in vitro bioactive properties. An ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry and multivariate statistics were combined to search for potential markers, in order to better discriminate the different commercial samples. Several phytochemicals were identified, namely alkaloids, phenolics, organosulfurs, and terpenoids. The polyherbal formulation Zigbir was the best source of phytochemicals, accounting for a cumulative total content of phytochemicals equal to 3.03 mg Eq./g, being particularly abundant in terpenoids, stilbenes, phenolic acids, and small-molecular-weight phenolics. Multivariate statistics allowed to group the different products in 2 bioactive subclusters. The diterpenoid andrographolide recorded the highest abundance in Zigbir and Sangrovit. The most predictive biomarkers were: piperine, isoquercitrin, 6-methylthiohexyldesulfoglucosinolate, 6-methylumbelliferone, benzoic acid, (+)-(1R,2R)-1,2-diphenylethane-1,2-diol, and piperitenone. Flavonoids were highly correlated with both in vitro antioxidant and enzyme inhibition assays. Our findings provide new insights into the comprehensive phytochemical composition of commercial feed additives and blend of botanical extracts used for both ruminant and non-ruminant nutrition. A great importance of polyphenols in relation to the biological activities was detected.


Sensor-based arm skill training in chronic stroke patients: results on treatment outcome, patient motivation, and system usability.
As stroke incidence increases, therapists' time is under pressure. Technology-supported rehabilitation may offer new opportunities. The objective of this study was to evaluate patient motivation for and the feasibility and effects of a new technology-supported task-oriented arm training regime (T-TOAT). Nine chronic stroke patients performed T-TOAT (2 x 30 min/day, four days/week) during eight weeks. A system including movement tracking sensors, exercise board, and software-based toolkit was used for skill training. Measures were recorded at baseline, after four and eight weeks of training, and six months posttraining. T-TOAT improved arm-hand performance significantly on Fugl-Meyer, Action Research Arm Test, and Motor Activity Log. Training effects lasted at least six months posttraining. Health-related-quality-of-life had improved significantly after eight weeks of T-TOAT with regard to perceived physical health, but not to perceived mental health (SF-36). None of the EuroQol-5D components showed significant differences before and after training. Participants were intrinsically motivated and felt competent to use the system. Furthermore, system usability was rated very good. However, exercise challenge as perceived by participants decreased significantly over eight weeks of training. The results of this study indicate that T-TOAT is feasible. Despite the small number of stroke patients tested, significant and clinically relevant improvements in skilled arm-hand performance were found.


Peptide array-based interactomics.
The analysis of protein-protein interactions (PPIs) is essential for the understanding of cellular signaling. Besides probing PPIs with immunoprecipitation-based techniques, peptide pull-downs are an alternative tool specifically useful to study interactome changes induced by post-translational modifications. Peptides for pull-downs can be chemically synthesized and thus offer the possibility to include amino acid exchanges and post-translational modifications (PTMs) in the pull-down reaction. The combination of peptide pull-down and analysis of the binding partners with mass spectrometry offers the direct measurement of interactome changes induced by PTMs or by amino acid exchanges in the interaction site. The possibility of large-scale peptide synthesis on a membrane surface opened the possibility to systematically analyze interactome changes for mutations of many proteins at the same time. Short linear motifs (SLiMs) are amino acid patterns that can mediate protein binding. A significant number of SLiMs are located in regions of proteins, which are lacking a secondary structure, making the interaction motifs readily available for binding reactions. Peptides are particularly well suited to study protein interactions, which are based on SLiM-mediated binding. New technologies using arrayed peptides for interaction studies are able to identify SLIM-based interaction and identify the interaction motifs.


Model of flexural fatigue damage accumulation for cortical bone.
Analytical models that predict modulus degradation in cortical bone subjected to uniaxial fatigue loading in tension and compression are presented. On the basis of experimental observations, damage was modeled as self-limiting for tension but not for compression. These mechanistic uniaxial damage models were then developed into a model for flexural fatigue of cortical bone based on laminated beam theory. The unknown coefficients in the uniaxial damage models were obtained by successfully fitting the resulting equations to uniaxial fatigue data from the literature on human cortical bone in tension and compression. Then, the predictions of the flexural model for the behavior of human cortical bone were compared with experimental results from a small but independent set of specimens tested at three different ranges of load in our laboratory. The behavior of the modulus degradation curves and the flexural fatigue lives of the specimens were in excellent agreement with the predictions of the model.


Peripherally driven low-threshold inhibitory inputs to lamina I local-circuit and projection neurones: a new circuit for gating pain responses.
Spinal lamina I is a key element of the pain processing system which relays primary afferent input to supraspinal areas. However, little is known about how the signal is modulated by its intrinsic network including local-circuit neurones (LCNs) and much less numerous anterolateral tract projection neurones (PNs). Here, we used whole-cell patch clamp recordings in an isolated spinal cord preparation to examine properties of identified LCNs (n = 85) and PNs (n = 73) in their functionally preserved local networks. Forty LCNs showed spontaneous rhythmic firing (2-7 Hz) at zero current injection, which persisted in the presence of blockers of fast synaptic transmission. In the remaining cases, most LCNs and PNs fired tonically in response to depolarizing current injections. We identified LCNs and PNs receiving low-threshold primary afferent-driven inhibitory inputs, which in many cases were disynaptic and temporally preceded classical high-threshold excitatory inputs. This direct inhibitory link between low-threshold afferents and PNs can function as a postsynaptic gate controlling the nociceptive information flow in the spinal cord. The LCNs were found to be integrated into the superficial dorsal horn network by their receipt of monosynaptic and disynaptic inputs from other lamina I and II neurones. One-third of LCNs and two-thirds of PNs tested responded to substance P application. Thus, substance P released by a noxious afferent stimulation may excite PNs in two ways: directly, and via the activation of presynaptic LCN circuitries. In conclusion, we have described important properties of identified lamina I neurones and their roles in a new circuit for gating pain responses.


Evaluation of subdermal biodegradable implants incorporating rifampicin as a method of drug delivery in experimental tuberculosis of guinea pigs.
Conventional chemotherapy of tuberculosis and leprosy requires rifampicin to be administered orally. The long period of treatment and adverse side effects of the drug lead to poor compliance. To overcome this, subdermal implants incorporating rifampicin in pure and micro-encapsulated forms with biodegradable material were used as a new drug delivery system in experimental tuberculosis of guinea pigs. Two experiments were performed with 45-mg and 100-mg drug implants. Progress of infection was followed at intervals by studying necropsy scores and weights of the organs of predilection and levels of the drug in the blood were determined. There was a constant and sustained release of the drug in therapeutic concentrations for 30 and 50 days until the implants were completely assimilated without causing any damage to the implant site. The importance of multiple implants at long intervals is discussed.


Modeling light trapping in nanostructured solar cells.
The integration of nanophotonic and plasmonic structures with solar cells offers the ability to control and confine light in nanoscale dimensions. These nanostructures can be used to couple incident sunlight into both localized and guided modes, enhancing absorption while reducing the quantity of material. Here we use electromagnetic modeling to study the resonances in a solar cell containing both plasmonic metal back contacts and nanostructured semiconductor top contacts, identify the local and guided modes contributing to enhanced absorption, and optimize the design. We then study the role of the different interfaces and show that Al is a viable plasmonic back contact material.


Prevalence of high screen time and associated factors among students: a cross-sectional study in Zhejiang, China.
To investigate the prevalence and correlates of high screen time (ST) among students in Zhejiang, China. Cross-sectional study. School-based adolescent health survey in Zhejiang Province, China. 23 543 students in grades 7-12 from 442 different schools. High ST. The mean age of the students was 15.6 years and 49.7% of them were girls. The prevalence of high ST (screen viewing ≥2 hours per day) was 42.4% (95% CI 40.2% to 44.5%), higher in boys than in girls (45.4%(95% CI 42.8% to 48.0%) vs 39.1% (95% CI 36.6% to 41.7%)). No statistically significant difference was found between urban and rural areas (43.0% (95% CI 37.2% to 48.7%) vs 42.1% (95% CI 39.6% to 44.6%)). The prevalence of high ST among middle school, academic high school and vocational high school students was 35.3%, 30.0% and 73.5%, respectively. Multivariable logistic analysis showed that older age, attendance at vocational high school, non-intact family, poor academic performance, bad self-reported health status, loneliness and drinking carbonated beverages ≥3 times every day were positively associated with high ST. Attendance at academic high school, higher parental education and being physically active were negatively associated with high ST. High ST was prevalent among students and associated with a cluster of sociodemographic and behavioural risk factors in Zhejiang, China.


Direct coupling of a free-flow isotachophoresis (FFITP) device with electrospray ionization mass spectrometry (ESI-MS).
We present the online coupling of a free-flow isotachophoresis (FFITP) device to an electrospray ionization mass spectrometer (ESI-MS) for continuous analysis without extensive sample preparation. Free-flow-electrophoresis techniques are used for continuous electrophoretic separations using an electric field applied perpendicular to the buffer and sample flow, with FFITP using a discontinuous electrolyte system to concurrently focus a target analyte and remove interferences. The online coupling of FFITP to ESI-MS decouples the separation and detection timeframe because the electrophoretic separation takes place perpendicular to the flow direction, which can be beneficial for monitoring (bio)chemical changes and/or extensive MS(n) studies. We demonstrated the coupling of FFITP with ESI-MS for simultaneous concentration of target analytes and sample clean-up. Furthermore, we show hydrodynamic control of the fluidic fraction injected into the MS, allowing for fluidically controlled scanning of the ITP window. Future applications of this approach are expected in monitoring biochemical changes and proteomics.


The 2010 outbreak of cholera among workers of a jute mill in Kolkata, West Bengal, India.
On 10 March 2010, an outbreak of diarrhoeal disease was reported among workers of a jute mill in Kolkata, West Bengal, India. The cluster was investigated to identify the agent(s) and the source of infection and make recommendations. A suspected case of cholera was defined as having >3 loose watery stools in a 24-hour period and searched for case-patients in the workers' colony. The outbreak was described by time, place, and person, and a case-control study was conducted to identify the source of infection. Rectal swabs were collected from the hospitalized case-patients, and the local water-supply system was assessed. In total, 197 case-patients were identified among 5,910 residents of the workers' colony (attack rate 3.33%). Fifteen of 24 stool samples were positive for Vibrio cholerae O1. The outbreak started on 7 March, peaked on 11 March, and ended on 16 March 2010. Compared to 120 controls, 60 cases did not differ in terms of age and socioeconomic status. Drinking-water from the reservoir within the mill premises was associated with an increased risk of illness [odds ratio: 26.7, 95% confidence interval (CI) 11.4-62.6) and accounted for most cases (population attributable risk percentage = 82%, 95% CI 70.8-92.9). An outbreak of cholera occurred among workers of the jute mill due to contamination of the drinking-water reservoir. It occurred within a few days of re-opening of the mill after the workers' strike. Health authorities need to enforce disinfection of drinking-water and regularly test its bacteriological quality, particularly before re-opening of the mill after the strike.


Hyperglycemia induces memory impairment linked to increased acetylcholinesterase activity in zebrafish (Danio rerio).
Diabetes mellitus, which causes hyperglycemia, affects the central nervous system and can impairs cognitive functions, such as memory. The aim of this study was to investigate the effects of hyperglycemia on memory as well as on the activity of acethylcholinesterase. Hyperglycemia was induced in adult zebrafish by immersion in glucose 111mM by 14 days. The animals were divided in 4 groups: control, glucose-treated, glucose-washout 7-days and glucose-washout 14-days. We evaluated the performance in inhibitory avoidance task and locomotor activity. We also determined acethylcholinesterase activity and gene expression from whole brain. In order to counteract the effect of hyperglycemia underlined by effects on acethylcholinesterase activity, we treated the animals with galantamine (0.05ng/g), an inhibitor of this enzyme. Also we evaluated the gene expression of insulin receptor and glucose transporter from zebrafish brain. The hyperglycemia promoted memory deficit in adult zebrafish, which can be explained by increased AChE activity. The ache mRNA levels from zebrafish brain were decrease in 111mM glucose group and returned to normal levels after 7 days of glucose withdrawal. Insulin receptors (insra-1, insra-2, insrb-1 and insrb-2) and glut-3 mRNA levels were not significantly changed. Our results also demonstrated that galantamine was able to reverse the memory deficit caused by hyperglycemia, demonstrating that these effects involve modulation of AChE activity. These data suggest that the memory impairment induced by hyperglycemia is underlined by the cholinergic dysfunction caused by the mechanisms involving the control of acetylcholinesterase function and gene expression.


The growth and the fluid dynamics of protein crystals and soft organic tissues: models and simulations, similarities and differences.
The fluid-dynamic environment within typical growth reactors as well as the interaction of such flow with the intrinsic kinetics of the growth process are investigated in the frame of the new fields of protein crystal and tissue engineering. The paper uses available data to introduce a set of novel growth models. The surface conditions are coupled to the exchange mass flux at the specimen/culture-medium interface and lead to the introduction of a group of differential equations for the nutrient concentration around the sample and for the evolution of the construct mass displacement. These models take into account the sensitivity of the construct/liquid interface to the level of supersaturation in the case of macromolecular crystal growth and to the "direct" effect of the fluid-dynamic shear stress in the case of biological tissue growth. They then are used to show how the proposed surface kinetic laws can predict (through sophisticated numerical simulations) many of the known characteristics of protein crystals and biological tissues produced using well-known and widely used reactors. This procedure provides validation of the models and associated numerical method and at the same time gives insights into the mechanisms of the phenomena. The onset of morphological instabilities is discussed and investigated in detail. The interplay between the increasing size of the sample and the structure of the convective field established inside the reactor is analysed. It is shown that this interaction is essential in determining the time evolution of the specimen shape. Analogies about growing macromolecular crystals and growing biological tissues are pointed out in terms of behaviours and cause-and-effect relationships. These aspects lead to a common source (in terms of original mathematical models, ideas and results) made available for the scientific community under the optimistic idea that the contacts established between the "two fields of engineering" will develop into an ongoing, mutually beneficial dialogue.


Artificial intelligence for diagnosis and Gleason grading of prostate cancer: the PANDA challenge.
Artificial intelligence (AI) has shown promise for diagnosing prostate cancer in biopsies. However, results have been limited to individual studies, lacking validation in multinational settings. Competitions have been shown to be accelerators for medical imaging innovations, but their impact is hindered by lack of reproducibility and independent validation. With this in mind, we organized the PANDA challenge-the largest histopathology competition to date, joined by 1,290 developers-to catalyze development of reproducible AI algorithms for Gleason grading using 10,616 digitized prostate biopsies. We validated that a diverse set of submitted algorithms reached pathologist-level performance on independent cross-continental cohorts, fully blinded to the algorithm developers. On United States and European external validation sets, the algorithms achieved agreements of 0.862 (quadratically weighted κ, 95% confidence interval (CI), 0.840-0.884) and 0.868 (95% CI, 0.835-0.900) with expert uropathologists. Successful generalization across different patient populations, laboratories and reference standards, achieved by a variety of algorithmic approaches, warrants evaluating AI-based Gleason grading in prospective clinical trials.


Association of slime with pathogenicity of coagulase-negative staphylococci causing nosocomial septicemia.
To assess the role of slime in the pathogenesis of nosocomial bloodstream infections caused by coagulase-negative staphylococci, we compared the characteristics of 27 nosocomial bloodstream isolates with those of 27 skin isolates from non-hospital personnel. Of 27 bloodstream isolates, 14 were judged to be significant by a clinical index, and 13 were contaminants. Slime production was observed in 13 of 14 significant isolates but in only 3 of 13 contaminants (P = 0.0003) and 4 of 27 skin isolates (P = 0.0001). The 14 pathogens were identified as Staphylococcus epidermidis. Only 7 of 13 contaminants and 9 of 27 skin isolates belonged to the same species (P less than 0.006). Slime-producing strains of S. epidermidis represented 13 of 14 pathogens but only 2 of 13 contaminants (P less than 0.0003). Neither adherence to Teflon catheters nor phagocytosis and killing of coagulase-negative staphylococci by polymorphonuclear leukocytes was significantly influenced by slime production. Nevertheless, the identity of the organism and the slime production test predicted the clinical significance of blood isolates of coagulase-negative staphylococci with an overall accuracy of 89%.


Is there a need for preoperative imaging of the internal mammary recipient site for autologous breast reconstruction?
Preoperative imaging of recipient-site vasculatur in autologous breast reconstruction may potentiate improved outcomes through the identification of individual variations in vascular architecture. There are a range of both normal and pathologic states which can substantially affect the internal mammary vessels in particular, and the identification of these preoperatively may significantly affect operative approach. There are a range of imaging modalities available, with ultrasound particularly useful, and computed tomography angiography (CTA) evolving as a useful option, albeit with radiation exposure. The benefits of CTA must be balanced against its risks, which include contrast nephrotoxicity and allergic reactions, and radiation exposure. The radiation risk with thoracic imaging is substantially higher than that for donor sites, such as the abdominal wall, with reasons including exposure of the contralateral breast to radiation (with a risk of contralateral breast cancer in this population 2 to 6 times higher than that of primary breast cancer, reaching a 20-year incidence of 15%), as well as proximity to the thyroid gland. Current evidence suggests that although many cases may not warrant such imaging because of risk, the benefits of preoperative CTA in selected patients may outweigh the risks of exposure, prompting an individualized approach.


With HIV prevalence among women increasing, more provinces encourage prenatal testing.
Testing of pregnant women for antibodies to HIV remains a thorny medical and ethical issue. On the one side are children who stand to benefit if their mothers' HIV status is determined. On the other is women's right to determine what tests they will receive. A majority of provinces now have counselling programs in place to encourage women to undergo testing. However, testing remains voluntary and informed consent is required before a test is done--policies that have CMA support.