The APG8/12-activating enzyme APG7 is required for proper nutrient recycling and senescence in Arabidopsis thaliana.
The vacuole/lysosome serves an important recycling function during starvation and senescence in eukaryotes via a process called autophagy. Here bulk cytosolic constituents and organelles become sequestered in specialized autophagic vesicles, which then deliver their cargo to the vacuole for degradation. In yeasts, genetic screens have identified two novel post-translational modification pathways remarkably similar to ubiquitination that are required for autophagy. From searches of the Arabidopsis genome, we have identified gene families encoding proteins related to both the APG8 and -12 polypeptide tags and orthologs for all components required for their attachment. A single APG7 gene encodes the ATP-dependent activating enzyme that initiates both conjugation pathways. Phenotypic analysis of an APG7 disruption indicates that it is not essential for normal growth and development in Arabidopsis. However, the apg7-1 mutant is hypersensitive to nutrient limiting conditions and displays premature leaf senescence. mRNAs for both APG7 and APG8 preferentially accumulate as leaves senesce, suggesting that both conjugation pathways are up-regulated during the senescence syndrome. These findings show that the APG8/12 conjugation pathways have been conserved in plants and may have important roles in autophagic recycling, especially during situations that require substantial nitrogen and carbon mobilization.


Conformational preferences in a benzodiazepine series of potent nonpeptide fibrinogen receptor antagonists.
Previously, we reported the direct design of highly potent nonpeptide 3-oxo-1,4-benzodiazepine fibrinogen receptor antagonists from a constrained, RGD-containing cyclic semipeptide. The critical features incorporated into the design of these nonpeptides were the exocyclic amide at the 8-position which overlaid the Arg carbonyl, the phenyl ring which maintained an extended Gly conformation, and the diazepine ring which mimicked the gamma-turn at Asp. In this paper, we investigate conformational preferences of the 8-substituted benzodiazepine analogues by examining structural modifications to both the exocyclic amide and the seven-membered diazepine ring and by studying the conformation of the benzodiazepine ring using molecular modeling, X-ray crystallography, and NMR. We found that the directionality of the amide at the 8-position had little effect on activity and the (E)-olefin analogue retained significant potency, indicating that the trans orientation of the amide, and not the carbonyl or NH groups, made the largest contribution to the observed activity. For the diazepine ring, with the exception of the closely analogous 3-oxo-2-benzazepine ring system described previously, all of the modifications led to a significant reduction in activity compared to the potent 3-oxo-1, 4-benzodiazepine parent ring system, implicating this particular type of ring system as a desirable structural feature for high potency. Energy minimizations of a number of the modified analogues revealed that none could adopt the same low-energy conformation as the one shared by the active (S)-isomer of the 3-oxo-1, 4-benzodiazepines and 3-oxo-2-benzazepines. The overall data suggest that the features contributing to the observed high potency in this series are the orientation of the 3-4 amide and the conformational constraint imposed by the seven-membered ring, both of which position the key acidic and basic groups in the proper spatial relationship.


Role of HERV-W syncytin-1 in placentation and maintenance of human pregnancy.
Over half of human genome contains retroelements, including retrotransposons, retroviruses, and other elements. Human endogenous retroviruses (HERVs) comprise about 8% of human genome. The products of 2 of 16 identified genes of HERV-W seem to play a pivotal role in the placentation. These 2 genes are HERV-W env glycoprotein (syncytin-1) and HERV-FRD env glycoprotein (syncytin-2). It has been shown previously that syncytin-1 mediates cell-cell fusions of cytotrophoblasts into syncytiotrophoblasts. In addition, HERV-W env contains an immunosuppressive region that may prevent rejection of a semiallogenic fetus from the mother's immune system. We analyzed 40 full-term placental tissues to localize the expression of syncytin-1-ISR by immunohistochemical staining and by reverse trancscriptase (RT) in situ polymerase chain reaction (PCR). Both the immunostaining and in situ RT-PCR showed strong expression of syncytin-1 in the syncytiotrophoblast layer from the full-term placental tissues. To further analyze the mechanism of early embryo HERV-W env activation, we utilized a HTR-8/SVneo cell line developed from first trimester human trophoblasts and subjected them to various physiologic concentrations of maternal hormones. Quantitative RT-PCR analyses demonstrated that exposure to progesterone significantly upregulated the HERV-W env expression, whereas several other hormones apparently played lesser roles. In conclusion, our findings suggest that expression of syncytin-1 (HERV-W env) in utero is expressed exclusively in the syncytiotrophoblast layer and is upregulated by progesterone.


Sexual selection in complex environments.
Sexual selection has resulted in some of the most captivating features of insects, including flashy colors, bizarre structures, and complex pheromones. These features evolve in dynamic environments, where conditions can change rapidly over space and time. However, only recently has ecological complexity been embraced by theory and practice in sexual selection. We review replicated selection studies as well as studies on variation in the agents of selection to delineate gaps in current knowledge and clarify exciting new directions for research. Existing work suggests that fluctuations in sexual selection may be extremely common, though work on the ecological factors influencing these fluctuations is scarce. We suggest that deeper ecological perspectives on sexual selection may alter some of the fundamental assumptions of sexual selection theory and rapidly lead to new discoveries.


Dynamics of polymer knots at equilibrium.
The relaxation and diffusion dynamics of knotted polymers at equilibrium under good solvent conditions are investigated by dynamic Monte Carlo simulations. Prime knots of chain lengths up to N=240 monomers and knots up to 20 essential crossings are studied. The relaxation dynamics of the prime knots at equilibrium do not display the classification into group as in the case of the nonequilibrium relaxation of cut knots [Phys. Rev. E 58, R1222 (1998); Phys. Rev. Lett. 87, 175503 (2001)]. Furthermore, the time autocorrelation functions for the radius of gyration of the nontrivial knots can be fitted by a sum of two exponential decays of long and short characteristic relaxation times. These two relaxation times decrease with the number of essential crossings C. The faster relaxation follows the Rouse behavior and scales as N1+2nu and its dependence on C is consistent with the scaling analysis using the blob picture. The mean-square displacement of the center of mass of the knots obeys the free diffusion behavior compatible with the Rouse dynamics. The diffusion coefficients of the knots, D approximately 1/N for large N, but D decreases for knots with increasing C. These results are analyzed using scaling theories and discussed in terms of topological interactions in the knots.


Adverse drug reactions of montelukast and pranlukast: Analysis of the Korea database.
Leukotriene receptor antagonists are recommended to treat asthma and allergic rhinitis. Although they had been used for a long time, recent studies have reported neuropsychiatric adverse drug reactions are associated with montelukast. This study analyzed the adverse drug reactions of montelukast and pranlukast, which are the two most frequently prescribed leukotriene receptor antagonists, respectively in Korea. This study retrospectively reviewed ADRs of 5,426 montelukast and 1,146 pranlukast reported in the Korea Adverse Event Reporting System between January 2014 and December 2018. When both drugs are classified by system organ class, the most adverse drug reactions were related to the gastro-intestinal system, followed by psychiatric events. The reported adverse drug reactions for both drugs were more common in women, and the ratio of adverse drug reactions to prescriptions was highest in the elderly. Women aged 19 to 64 years reported more than twice as many adverse drug reactions than men of the same age, and more than 5 times in insomnia. When prescribing montelukast and pranlukast, attention would need to digestive and sleep disorders, especially women aged 19 to 64. After prescribing montelukast, physicians would need to pay more attention to agitation (5/396378 vs 0/82475), bad or vivid dreams (6/396378 vs 0/82475), anxiety (11/396378 vs 0/82475), depression (14/396378 vs 1/82475), tremor (53/396378 vs 7/82475), irritability (5/396378 vs 1/82475), insomnia (159/396378 vs 25/82475), and headache (68/396378 vs 10/82475), compared to when prescribing pranlukast. Further prospective research needs to elucidate the relationship between neuropsychiatric events and montelukast.


Septic induced abortions.
Induced abortions are not infrequent in our community. Being the only tertiary level care hospital in Hazara division, Ayub Teaching Hospital frequently receives cases of induced abortions with complications attempted by inappropriately trained personnel. It was decided to study the frequency of such cases in our admitted patients, complications and their outcome in such risky situation. A cross sectional study was conducted from January 2006 to December 2007 in Gynae-B Unit, Ayub Teaching Hospital, Abbottabad. All admitted patients in the unit in the 1st trimester of pregnancy and handling outside the hospital were included in the study. In two years period total Gynae admissions were 1090. There were 152 (13.95%) cases of spontaneous 1st trimester abortions and 52 (4.7%) cases of induced abortions which are included in the study. Age range of the patients was between 20-45 years, with a mean of 35 years. Majority of the patients were grand multigravidas belonging to lower socioeconomic group and not using any contraception. Twenty patients (38.4%) presented with heavy per vaginal bleeding due to incomplete nature of the procedure. Twelve patients 23% presented in shock due to excessive per vaginal bleeding. Ten patients 19.2% had uterine perforation and intraperitoneal haemorrhage. Two patients 3.8% had gut injury associated with uterine perforation. Three patients 5.7% developed septicaemia due to uterine gangrene. Ten patients 19.2% had acute pelvic infection presented with pelvic abscess and acute pelvic inflammatory disease. Maternal motility in our study was 2 out of 52 (3.8%). Induced septic abortions contribute significantly to maternal morbidity and mortality. Improving literacy rate in our female population and effective family planning should reduce its incidence. Different resources should be used to develop awareness of the hazards of induced abortions in the community.


Inflight exercise affects stand test responses after space flight.
The purpose of this study was to determine whether exercise performed by Space Shuttle crew members during short-duration space flights (9-16 d) affects the heart rate (HR) and blood pressure (BP) responses to standing within 2-4 h of landing. Thirty crew members performed self-selected inflight exercise and maintained exercise logs to monitor their exercise intensity and duration. Two subjects participated in this investigation during two different flights. A 10-min stand test, preceded by at least 6 min of quiet supine rest, was completed 10-15 d before launch (PRE) and within 4 h of landing (POST). Based upon their inflight exercise records, subjects were grouped as either high (HIex: > or = 3 times/week, HR > or = 70% HRmax, > or = 20 min/session, N = 11), medium (MEDex: > or = 3 times/week, HR < 70% HRmax, > or = 20 min/session, N = 10), or low (LOex: < or = 3 times/week, HR and duration variable, N = 11) exercisers. HR and BP responses to standing were compared between groups (ANOVA, P < or = 0.05). There were no PRE differences between the groups in supine or standing HR and BP. Although POST supine HR was similar to PRE, all groups had an increased standing HR compared with PRE. The increase in HR upon standing was significantly greater after flight in the LOex group (36 +/- 5 bpm) compared with HIex or MEDex groups (25 +/- 1 bpm; 22 +/- 2 bpm). Similarly, the decrease in pulse pressure (PP) from supine to standing was unchanged after space flight in the MEDex and HIex groups but was significantly greater in the LOex group (PRE: -9 +/- 3; POST: -19 +/- 4 mm Hg). Thus, moderate to high levels of inflight exercise attenuated HR and PP responses to standing after space flight.


Bioactive natural products from orchids native to the Americas - A review.
The purpose of this review is to provide information on the traditional uses, phytochemical and pharmacological studies performed with species of orchids native to the Americas and the Caribbean Islands. The treatment of inflammation is the most traditional use for plants of this family, specially in Central America, while anti-inflammatory and anticancer assays are oftenly reported in pharmacological investigations. From the chemical point of view, they are sources of phenanthrenoids and stilbenes, rare secondary metabolites not commonly found in other families of plants, as well as cycloartane triterpenes, pyrrolizidine alkaloids and flavonoids. Since just few species were chemically and pharmacologically studied, in comparison to the large number of native species (less than 0.5% of the total), the orchids of the New World may be an interesting niche for the discovery of new, bioactive natural products.


Polyelectrolyte complex materials consisting of antibacterial and cell-supporting layers.
The characterization of a polyelectrolyte complex material comprised of two biopolymers, a chitosan upper layer and a gellan gum under layer, is reported. It is shown that the upper layer of chitosan with incorporated levofloxacin displays an antibacterial activity, while the under layer of a gellan gum/TiO(2) composite supports the growth of fibroblastic cells.


Redirecting RNA splicing by SMAD3 turns TGF-β into a tumor promoter.
Transforming growth factor β (TGF-β) is a well-known growth inhibitor of normal epithelial cells, but it is also secreted by solid tumors to promote cancer progression. Our recent discovery of SMAD3-PCBP1 complex with direct RNA-binding properties has shed light on how this conversion is implemented by controlling pre-mRNA splicing patterns.


Molecular inhibition of angiogenesis and metastatic potential in human squamous cell carcinomas after epidermal growth factor receptor blockade.
Tumor metastasis represents a complex multistep process that requires migration, invasion, and angiogenesis. In this study, we examined the impact of molecular blockade of the epidermal growth factor receptor on the invasive and metastatic capacity of human squamous cell carcinoma (SCC) of the head and neck using in vitro and in vivo model systems. Treatment with the anti-epidermal growth factor receptor antibody C225 attenuated the migration of SCC-1 tumor cells through a chemotaxis chamber in a dose-dependent manner. Incubation of SCC cells with 10-100 nM C225 for 4 h resulted in 40-60% inhibition of cell migration. Furthermore, in the presence of C225, the capacity of SCC-1 to invade across a layer of extracellular matrix (Matrigel) was significantly inhibited. Using an in vivo orthotopic floor-of-mouth xenograft model, locoregional tumor invasion of SCC-1 into muscle, vessel, bone, and perineural tissues was inhibited in C225-treated mice. This inhibition was additionally characterized by down-regulation in the expression of matrix metalloproteinase-9. These data suggest that inhibition of metastatic potential by C225 may be mediated via decreased migration and invasion of SCC cells. Regarding angiogenesis in vitro, we first studied human umbilical vascular endothelial cells, which established a capillary-like network structure (tube formation) in the presence of reconstituted Matrigel. Treatment with C225 reduced cell-to-cell interaction of human umbilical vascular endothelial cells, resulting in disruption of tube formation. The effect of C225 was additionally examined using an in vivo tumor xenograft neovascularization model of angiogenesis. Systemic treatment with C225 not only reduced tumor growth and the number of blood capillaries but also hindered the growth of established vessels toward the tumor. Taken together, these results provide evidence that C225 can suppress tumor-induced neovascularization and metastasis in SCC of the head and neck.


Conventional apheresis therapies: a review.
This article reviews advances in the scientific basis and medical practice of plasmapheresis and cytapheresis therapies. Newly-characterized autoantibodies in neuromyelitis optica, Guillain-Barre variants, anti-neutrophil cytoplasmic antibody (ANCA) vasculitides, etc., exemplify the modern molecular biology which now provides a rigorous framework of understanding for the clinical practice of plasmapheresis. Clinical trials continue to clarify the appropriate use of therapeutic plasmapheresis (TPE) in these and other diseases. Centrifugal (cTPE) and membrane filtration (mTPE) types of plasmapheresis are compared, with details of the plasmapheresis prescription, anticoagulation choices, replacement fluids and other practical considerations. Plasma removal is more efficient with cTPE; mTPE systems have a lower plasma extraction ratio, and therefore require higher blood flow rates or longer procedure times. Autoantibodies and other pathogenic macromolecules targeted for removal by plasmapheresis can be depleted predictably when the plasma is discarded, as in conventional TPE. On-line plasma processing to regenerate the patient's own plasma avoids the need for replacement albumin solutions or plasma transfusion, but is inherently less efficient at removing the target molecule, so usually requires a longer procedure. Therapeutic white cell reduction (leukapheresis), platelet reduction (thrombocytapheresis) and red cell exchange (erythrocytapheresis) require centrifugal apheresis systems.


Endocervical polyps in high risk human papillomavirus infections.
Human papillomavirus (HPV) positive patients with and without endocervical polyps is compared with respect to HPV genotypes and presence of pre-invasive diseases. To our knowledge, this is the first and largest report in the literature examining the endocervical polyps in HPV positive cases. Clinicopathological data for the first one million screening patients (n = 1060 992) from around the entire country during 2015 and 2016 were targeted for this research. Colposcopy, colposcopic surgical diagnostic procedures and final pathology results of 3499 patients with high-risk (HR) HPV-positive were obtained from reference colposcopy centers. Patients with endocervical polyps (n = 243 [6.9 %]) were accepted as experimental arm while patients without any endocervical polyp (n = 3256 [93.1%]) were regarded as the control group. Age, HPV genotype, Pap smear abnormality, and final pathological results were compared between two groups using Student's t-test and cross-tabulation chi-square test. The incidence of endocervical polyp was found to be 6.9 % in HR HPV-positive women. The most common HPV genotypes observed in both groups were HPV 16 or 18. Abnormal cytology reports (≥ ASC-US) were not significantly different between both groups. However, with respect to final pathological diagnosis, patients with endocervical polyp had significantly lower numbers of pre-invasive diseases (31.3% vs 44.2%; p < 0.10). Endocervical polyps may be more common in patients with HR HPV infections. HPV 18 is observed significantly more, in the HR HPV positive endocervical polyp group. Patients with endocervical polyps do not have increased risk for preinvasive cervical diseases.


Sex Chromosomes and Master Sex-Determining Genes in Turtles and Other Reptiles.
Among tetrapods, the well differentiated heteromorphic sex chromosomes of birds and mammals have been highly investigated and their master sex-determining (MSD) gene, Dmrt1 and SRY, respectively, have been identified. The homomorphic sex chromosomes of reptiles have been the least studied, but the gap with birds and mammals has begun to fill. This review describes our current knowledge of reptilian sex chromosomes at the cytogenetic and molecular level. Most of it arose recently from various studies comparing male to female gene content. This includes restriction site-associated DNA sequencing (RAD-Seq) experiments in several male and female samples, RNA sequencing and identification of Z- or X-linked genes by male/female comparative transcriptome coverage, and male/female transcriptomic or transcriptome/genome substraction approaches allowing the identification of Y- or W-linked transcripts. A few putative master sex-determining (MSD) genes have been proposed, but none has been demonstrated yet. Lastly, future directions in the field of reptilian sex chromosomes and their MSD gene studies are considered.


[Development and application of separation materials for mixed-mode chromatography].
Mixed-mode chromatography has received more and more attention due to its unique chromatographic characteristics recently. In this review, a summary of the development and applications of mixed-mode chromatography is presented. According to the types of hydrophilic interaction/ion-exchange mixed-mode chromatography (HILIC/IEX), reversed-phase/hydrophilic interaction mixed-mode chromatography (RPLC/HILIC) and reversed-phase/ion-exchange mixed-mode chromatography (RPLC/IEX), the preparation and applications of each type are introduced. Compared with single mode chromatography, the selectivity as well as the sample loading capacity of the mixed-mode chromatography can be improved. The researches mostly focus on the design of mixed-mode stationary phases and the applications especially to bio-analysis. The mixed-mode chromatography will be a potent approach for the analysis and separation of complex samples, and it will also be an effective and complementary tool in practical separation work.


Reconstruction and analysis of genome-scale metabolic model for thermophilic fungus Myceliophthora thermophila.
Myceliophthora thermophila, a thermophilic fungus that can degrade and utilize all major polysaccharides in plant biomass, has great potential in biotechnological industries. Here, the first manually curated genome-scale metabolic model iDL1450 for M. thermophila was reconstructed using an autogenerating pipeline with thorough manual curation. The model contains 1450 genes, 2592 reactions, and 1784 unique metabolites. High accuracy was shown in predictions related to carbon and nitrogen source utilization based on data obtained from Biolog experiments. Besides, metabolism profiles were analyzed using iDL1450 integrated with transcriptomics data of M. thermophila at various growth temperatures. The refined model provides new insights into thermophilic fungi metabolism and sheds light on model-driven strain design to improve biotechnological applications of this thermophilic lignocellulosic fungus.


Gene therapy using bacterial vectors.
Bacteria can be used for gene therapy via two strategies - either by transfection of eukaryotic host cells using bacteria (bactofection) or by alternative gene therapy that does not alter the host genome, but uses the prokaryotic expression system, which can be controlled or stopped from outside. While bactofection is optimal for gene substitution and DNA vaccination, alternative gene therapy is suitable for in situ delivery of proteins and treatment with intracellular bactochondria. A specific form of bacteria-mediated gene therapy is transkingdom RNA interference. In this review, advantages and issues related to bacterial vectors as well as the major applications in biomedical research are summarized. Despite numerous published experiments, especially in the treatment of solid tumors and gut infections, progress in the clinics lags behind and major improvements in the safety, and even more in the efficiency, of these approaches are needed.


Opposite effects of electrostatics and steric exclusion on bundle formation by F-actin and other filamentous polyelectrolytes.
A number of positively charged polypeptides and proteins bundle DNA, F-actin, microtubules, and viruses such as filamentous phage fd and tobacco mosaic virus (TMV), as well as intermediate filaments formed by vimentin. The general behavior is dictated by the common polyelectrolyte nature of these biopolymers, which gives rise to nonspecific binding by ligands carrying several net opposite charges. An attractive interaction accounts for the subsequent lateral aggregation, distinguishing this transition from the liquid crystalline formation of filamentous particles at high concentrations. Morphologically similar filament bundles can also be induced by inert solutes such as polyethylene glycol (PEG) and proteins that do not bind the macromolecular filaments, but the physicochemistry underlying this class of bundle transitions is distinct. In particular, bundling transitions induced by electrostatic and steric mechanisms have an opposite dependence on the solution ionic strength and the concentration of the filamentous biopolymers. The distinct mechanisms illustrated in this report may each contribute to the formation of specific polymer bundles under physiological conditions.


Initiation of neuronal differentiation requires PI3-kinase/TOR signalling in the vertebrate neural tube.
Regulated neuron production within the vertebrate nervous system relies on input from multiple signalling pathways. Work in the Drosophila retina has demonstrated that PI3-kinase and downstream TOR signalling regulate the timing of photoreceptor differentiation; however, the function of such signals during vertebrate neurogenesis is not well understood. Here we show that mutant mice lacking PKB activity downstream of PDK1, the master kinase of the PI3-kinase pathway, exhibit deficient neuron production. We further demonstrate expression of PI3-kinase signalling components and active PKB and TOR signalling in the chick spinal cord, an early site of neurogenesis. Neuron production was also attenuated in the chick neural tube following exposure to small molecule inhibitors of PI3-kinase (LY294002) or TOR (Rapamycin) activity. Furthermore, Rapamycin repressed expression of early neuronal differentiation genes, such as Ngn2, but did not inhibit expression of Sox1B genes characteristic of proliferating neural progenitors. In addition, some cells expressing an early neuronal marker were mis-localised at the ventricular surface in the presence of Rapamycin and remained aberrantly within the cell cycle. These findings suggest that TOR signalling is necessary to initiate neuronal differentiation and that it may facilitate coordination of cell cycle and differentiation programmes. In contrast, stimulating PI3-kinase signalling did not increase neuron production, suggesting that such activity is simply permissive for vertebrate neurogenesis.


DNA methylation analysis of the HIF-1α prolyl hydroxylase domain genes PHD1, PHD2, PHD3 and the factor inhibiting HIF gene FIH in invasive breast carcinomas.
Hypoxia-inducible factor-1 (HIF-1) activity is regulated by prolyl hydroxylase (PHD1, PHD2, PHD3) and factor inhibiting HIF-1 (FIH) that target the α subunit of HIF-1 (HIF-1α) for proteosomal degradation. We hypothesised that the elevated HIF-1α level is due in some tumours to epigenetic silencing by DNA hypermethylation of the promoter region of one or more of the PHDs and FIH genes. The aims were to define the presence or absence of promoter methylation of PHDs and FIH in cell lines of various sources and breast carcinomas and, if present, determine its effect on mRNA and protein expression. Tumour cell lines (n = 20) and primary invasive breast carcinomas (n = 168) were examined for promoter region DNA methylation using methylation-sensitive high-resolution melting. There was evidence of PHD3 but not of PHD1, PHD2 or FIH DNA methylation in breast cancer (SkBr3) and leukaemic (HL60 and CCRF-CEM) cell lines, but there was no evidence of methylation in any of 168 breast cancers. Only the high-level PHD3 methylation seen in leukaemic cell lines correlated with absent mRNA and protein expression. Methylation-induced epigenetic silencing of PHD1, PHD2, PHD3 and FIH is unlikely to underlie up-regulated HIF-1α expression in human breast cancer but may play a role in other tumour types.


Kinetic assay of fluorescein mono-beta-D-galactoside hydrolysis by beta-galactosidase: a front-face measurement for strongly absorbing fluorogenic substrates.
A novel enzymatic assay method was developed for fluorogenic substrates that have significant intrinsic absorbance and fluorescence under the assay conditions. Fluorescein mono-beta-D-galactoside (FMG) was chosen as the substrate for the fluorescence enzymatic assay because of the high fluorescence of its hydrolytic product (fluorescein) and suitability of being hydrolyzed by beta-galactosidase. The fluorescence-concentration relationships for fluorescein and for FMG in both the right-angle detection mode of a fluorometer and the front-face detection mode of a fluorescence plate reader were exactly established and used to determine the kinetics of the enzyme assay. The results show that only front-face detection in the fluorescence plate reader can overcome the fluorescence concentration quenching that inevitably results from high absorbance by the intrinsically absorbing substrate in the conventional fluorometer, which utilizes right-angle detection. Only with front-face detection was the fluorescent assay of FMG hydrolysis under conditions of high optical density possible. The enzymatic measurements on the fluorescence plate reader were particularly efficient for determination of the enzyme kinetics because of the high rate of data collection. In this assay system, Michaelis-Menten constant Km and enzymatic catalysis rate k2 of FMG were determined as 117.6 microM and 22.7 mumol-(min.mg)-1, respectively. The results and methods described in this paper can be generalized for any assay using a fluorogenic substrate whether or not it has a high background absorbance.


Efficient Reaction Space Exploration with ChemTraYzer-TAD.
The development of a reaction model is often a time-consuming process, especially if unknown reactions have to be found and quantified. To alleviate the reaction modeling process, automated procedures for reaction space exploration are highly desired. We present ChemTraYzer-TAD, a new reactive molecular dynamics acceleration technique aimed at efficient reaction space exploration. The new method is based on the basin confinement strategy known from the temperature-accelerated dynamics (TAD) acceleration method. Our method features integrated ChemTraYzer bond-order processing steps for the automatic and on-the-fly determination of the positions of virtual walls in configuration space that confine the system in a potential energy basin. We use the example of 1,3-dioxolane-4-hydroperoxide-2-yl radical oxidation to show that ChemTraYzer-TAD finds more than 100 different parallel reactions for the given set of reactants in less than 2 ns of simulation time. Among the many observed reactions, ChemTraYzer-TAD finds the expected typical low-temperature reactions despite the use of extremely high simulation temperatures up to 5000 K. Our method also finds a new concerted β-scission plus O2 addition with a lower reaction barrier than the literature-known and so-far dominant β-scission.


Methodological considerations in pediatric behavioral research: measurement.
Measurement is the critical building block underlying all science. Adequate measures should demonstrate reliability, validity, and sensitivity. Special problems associated with the selection and development of measures used in behavioral pediatric research are reviewed. These include measurement specificity, construct complexity, the illusive "gold standard," construct independence, theoretical clarity, timing of measurements, and developmental considerations. The advantages and limitations of a variety of behavioral assessment strategies are described: questionnaires, interviews, behavioral observations, permanent products, biological measures, and provider ratings. Programmatic measurement research not only improves the science of behavioral pediatrics but also has widespread clinical application.


A case of congenital left ventricular diverticulum with pulmonary stenosis and its scintigraphic characteristics.
We encountered a 31-year-old female patient with mild valvular pulmonary stenosis who had no abnormality in the electrocardiogram but pulmonary dilatation in the chest radiograph. Two-dimensional echocardiography and magnetic resonance imaging demonstrated two small protrusions at the interventricular septum indicating diverticula. Large perfusion defects were observed at the anterior wall in the thallium-201 myocardial tomograms. Short axial and vertical long axial images by ECG-gated blood pool tomography revealed an out-pouching best seen during diastole and a good contraction during systole in the corresponding areas. These findings suggested the presence of thin but normal myocardium in the anterior wall, i.e. a muscular type of left ventricular diverticulum. The presence of the muscular type of left ventricular diverticulum at the anterior and septal walls was confirmed by contrast left ventriculography. A congenital diverticulum at the anterior and septal walls with pulmonary stenosis is very rare. Furthermore, its scintigraphic images were quite characteristic and useful for its diagnosis.


Self-Report and Biological Indexes of Work-Related Stress in Neonatal Healthcare Professionals: A Repeated-Measures Observational Study.
Healthcare providers working in neonatal intensive care units (NICUs) are considered at high risk for psychological work-related stress. To evaluate both perceived and biological measures of work-related stress in neonatal healthcare professionals and to compare professionals working in the NICU with their colleagues working in less critical environments (ie, neonatal wards [NWs]). The salivary cortisol level at the beginning (CORT-B) and at the end (CORT-E) of a daily work shift was collected once a week for 6 weeks and a psychological questionnaire was submitted to NW and NICU workers of a tertiary university center. No differences emerged in the overall cortisol secretion between professionals (NW 45 vs NICU 28), but the decrease in the mean cortisol values between CORT-B and CORT-E was less pronounced in NICU professionals (P < .001) who had greater psychological stress (P < .001). Lack of correlation between perceived and biological indexes was observed. NICU professionals reported greater levels of self-perceived psychological stress, especially in terms of professional self-doubt and the complexity of interactions with infants and their parents.The disconnection between psychological and biological indexes raises the issue that work-related stress might be covert to the professionals themselves. Dedicated resources should be developed to address quality of life and the work environment of NICU professionals. The absence of a correlation between perceived and biological indexes highlights the need to incorporate multidimensional physiological and biological measurements in evaluating burnout levels in neonatal healthcare providers.


Upbeat nystagmus in a patient with a small medullary infarct.
A 43 year old man presented with decreased sensitivity in the left side of the face and both upper limbs, and with upbeat nystagmus and skew deviation. MRI demonstrated a well defined lesion compatible with an infarct in the left side of the medulla, caudal and ventral to the vestibular nuclei, possibly involving the most caudal of the perihypoglossal nuclei, the nucleus intercalatus. Ocular motor studies, using an infrared system, showed that the nystagmus slow phase decayed exponentially, suggesting a failure of integration for vertical eye movements. Vertical integration might, therefore, be performed partly in the nucleus intercalatus.


Intercellular communication-filling in the gaps.
Coordination and synchrony of a variety of cellular activities in tissues of plants and animals occur as a consequence of the transfer of low molecular weight biosynthetic and signaling molecules through specialized structures (plasmodesmata in plant cells and gap junctions in mammalian cells) that form aqueous channels between contacting cells. Investigations with rat liver demonstrated that cell-cell communication is mediated by a 32 kilodalton polypeptide that forms a hexameric pore structure in the plasma membrane. Following association with the same structure in a contiguous cell, a trans-double membrane channel is created that has been termed a gap junction. In plant tissue, long tubelike structures called plasmodesmata are suggested to serve a similar cell-cell linking function between cytoplasmic compartments. Although morphologically distinct, dynamic observations suggest similarities in transport properties between gap junctions and plasmodesmata. Recent work now provides evidence that these functional similarities may reflect a more profound identity between the paradigm animal gap junction polypeptide (32 kilodalton rat liver polypeptide) and an immunologically homologous protein localized to plant plasma membrane/cell wall fractions that may be a component of plasmodesmata.


PHLS works towards effective interaction with primary care.
The PHLS primary care initiative was set up to develop and maintain links with primary care professionals, including researchers, GP research networks, and primary care groups in order to prioritize research and laboratory service developments. Such collaboration has spawned the development of antibiotic guidance, which has been posted on the Internet, and participation of the PHLS in other infectious disease guidance (for example, Prodigy). The implementation of the NHS information strategy will further facilitate the development of and access to local and national primary care guidance, laboratory reporting, and participation in surveillance. Primary care professionals identified several main research priorities in an exercise that facilitated collaboration in the development of research projects and grant applications. These priorities are the natural history of infections presenting to GPs, antibiotic resistance, the role of clinical scores and the laboratory in the diagnosis of infection, the importance of patient and professional attitudes in modifying antimicrobial prescribing, and the role of health professionals in community infection control. Current rapid changes in primary care in the new NHS--such as NHS Direct and the establishment of walk-in centres--may affect the surveillance of communicable disease by offering opportunities for improved data collection but also by posing threats to current systems of surveillance.


Mechanisms of cardiac hypertrophy and injury--possible role of protein kinase C activation.
To examine the molecular mechanisms by which mechanical stimuli induced cardiac hypertrophy and injury, we cultured rat neonatal cardiocytes in deformable dishes and imposed an in vitro mechanical load by stretching the adherent cells. Myocyte stretching increased total cell RNA content and mRNA levels of c-fos. Marked accumulation of c-fos mRNA followed increases in intracellular Na+ and protein kinase C activation. The accumulation of c-fos mRNA by cardiocyte stretching was suppressed by protein kinase C inhibitors but not by stretch channel blockers. Moreover, myocyte stretching increased inositol phosphate levels, and activation of protein kinase C by phorbolesters stumulated the expression of c-fos. We also examined TGF beta expression in the heart. TGF beta is known to be stimulated by protein kinase C activation, and the mRNA level of TGF beta was increased in in vivo heart by pressure overload. Furthermore, collagen synthesis was stimulated by TGF beta in cultured fibroblasts from hearts. These findings suggest that hemodynamic overload may stimulate cardiac hypertrophy and induce cardiac injury (fibrosis) through protein kinase C activation.


Prevalence of pancreatic diabetes in patients carrying mutations or polymorphisms of the PRSS1 gene in the Han population.
This study updated the estimated prevalence of type 3c diabetes damage to the pancreas through different genotypes of PRSS1 and their clinical characteristics in the Han population. Cross-sectional analysis was performed of the most recent (2003-2007) patients with pancreatitis from six hospitals of the Han population in South China (n = 253). There were 32 patients with pancreatitis carrying a PRSS1 gene abnormality within intron region among 253 cases of pancreatitis, including 27 patients carrying novel single nucleotide polymorphisms, namely, IVS 3 +75 A --> G conversion, and five patients with the mutation IVS3 + 10 T --> G. Among these patients, there were only three cases of patients with diabetes (9.37%). This was lower than the prevalence of abnormalities in the exons of the PRSS1 gene (51.92%): 12 patients with c.361 G --> A, eight patients with c.415 T --> A, and five patients with c.365G --> A. Among them were 12 persons with diabetes, including five requiring insulin to regulate blood sugar. What is more, among the 27 patients carrying PRSS1 gene polymorphism (c.486 C --> T, within the exon 4), there were 15 persons with diabetes symptoms. More than 40% of these patients required insulin to regulate blood sugar. An abnormality within the intron region of the PRSS1 gene represents one of the causes of pancreatitis in Chinese patients, but it is not related to pancreatic diabetes. However, the exon abnormality obviously raises the morbidity rate of type 3c diabetes, which relies on insulin.


DIGE Analysis of Fish Tissues.
Two-dimensional difference gel electrophoresis (2D-DIGE) appears to be especially useful in quantitative approaches, allowing the co-separation of proteins of control samples from proteins of treatment/disease samples on the same gel, eliminating gel-to-gel variability. The principle of 2D-DIGE is to label proteins prior to isoelectric focusing and use three spectrally resolvable fluorescent dyes, allowing the independent labeling of control and experimental samples. This procedure makes it possible to reduce the number of gels in an experiment, allowing the accurate and reproducible quantification of multiple samples. 2D-DIGE has been found to be an excellent methodical tool in several areas of fish research, including environmental pollution and toxicology, the mechanisms of development and disorders, reproduction, nutrition, evolution, and ecology.


Management of mental health conditions in refugee youth: An overview for the psychiatric-mental health nurse practitioner.
In 2017, there were a total of 70 million forcibly displaced individuals or refugees, worldwide. Unfortunately, over 50% of refugees are under the age of 18 years. Refugee youth are at-risk for a variety of mental health conditions. The purpose of this report is to provide psychiatric-mental health nurse practitioners (PMHNPs) with an understanding of unique cultural implications in refugee youth, risk factors towards the development of psychiatric illness, and means to identify those at-risk for sequelae or those meeting diagnostic criteria. Also presented are foundational treatment and management implications to ensure the provision of culturally sensitive and efficacious care to this unique population. Utilizing PubMed, CINAHL, and PsycINFO, a literature review was conducted to identify relevant publications pertaining to mental health issues in refugee youth spanning from 2013 to present. Reference lists of identified articles were also searched. A variety of risk and protective factors are discussed, spanning from premigration, during flight, and postsettling periods. PMHNPs must be proficient in screening and diagnosis of mental health conditions in refugee youth and implementation of pharmacological and psychotherapeutic interventions. PMHNPs must also be well versed in community-based resources that can be utilized to promote optimal outcomes.


Photo- and Electrocatalytic Reduction of CO2 over Metal-Organic Frameworks and Their Derived Oxides: A Correlation of the Reaction Mechanism with the Electronic Structure.
A Ce/Ti-based bimetallic 2-aminoterephthalate metal-organic framework (MOF) was synthesized and evaluated for photocatalytic reduction of CO2 in comparison with an isoreticular pristine monometallic Ce-terephthalate MOF. Owing to highly selective CO2 adsorption capability, optimized band gaps, higher flux of photogenerated electron-hole pairs, and a lower rate of recombination, this material exhibited better photocatalytic reduction of CO2 and lower hydrogen evolution compared to Ce-terephthalate. Thorough probing of the surface and electronic structure inferred that the reducibility of Ce4+ to Ce3+ was due to the introduction of an amine functional group into the linker, and low-lying Ti(3d) orbitals in Ce/Ti-2-aminoterephthalate facilitated the photoreduction reaction. Both the MOFs were calcined to their respective oxides of Ce1-xTixO2 and CeO2, and the electrocatalytic reduction of CO2 was performed over the oxidic materials. In contrast to the photocatalytic reaction mechanism, the lattice substitution of Ti in the CeO2 fluorite cubic structure showed a better hydrogen evolution reaction and consequently, poorer electroreduction of CO2 compared to pristine CeO2. Density functional theory calculations of the competitive hydrogen evolution reaction on the MOF and the oxide surfaces corroborated the experimental findings.


δEF1 associates with DNMT1 and maintains DNA methylation of the E-cadherin promoter in breast cancer cells.
Abnormal DNA methylation at the C-5 position of cytosine (5mC) of CpG dinucleotides is a well-known epigenetic feature of cancer. Levels of E-cadherin, which is regularly expressed in epithelial tissues, are frequently reduced in epithelial tumors due to transcriptional repression, sometimes accompanied by hypermethylation of the promoter region. δEF1 family proteins (δEF1/ZEB1 and SIP1/ZEB2), key regulators of the epithelial-mesenchymal transition (EMT), suppress E-cadherin expression at the transcriptional level. We recently showed that levels of mRNAs encoding δEF1 proteins are regulated reciprocally with E-cadherin level in breast cancer cells. Here, we examined the mechanism underlying downregulation of E-cadherin expression in three basal-type breast cancer cells in which the E-cadherin promoter region is hypermethylated (Hs578T) or moderately methylated (BT549 and MDA-MB-231). Regardless of methylation status, treatment with 5-aza-2'-deoxycytidine (5-aza), which inhibits DNA methyltransferases, had no effect on E-cadherin expression. Knockdown of δEF1 and SIP1 resulted in recovery of E-cadherin expression in cells lacking hypermethylation, whereas combined treatment with 5-aza synergistically restored E-cadherin expression, especially when the E-cadherin promoter was hypermethylated. Moreover, δEF1 interacted with DNA methyltransferase 1 (DNMT1) through the Smad-binding domain. Sustained knockdown of δEF1 family proteins reduced the number of 5mC sites in the E-cadherin promoter region, suggesting that these proteins maintain 5mC through interaction with DNMT1 in breast cancer cells. Thus, δEF1 family proteins appear to repress expression of E-cadherin during cancer progression, both directly at the transcriptional level and indirectly at the epigenetic level.


Aetiological, modifying and lethal factors in cleft lip and palate.
Prenatal factors influencing cleft lip (CL), cleft lip and palate (CLP) and isolated cleft palate (CP) may account for the development of the defect (aetiological factor) for a change in the degree or type of the defect (modifying factor), or for the death of the embryo (lethal factor). Each active factor appears to act in all three directions, all be it at different ratios. This is used for analysis of groups of cleft-effected individuals registered at the Department of Plastic Surgery, Prague, from the catchment area of Bohemia. In terms of cleft defect teratogenesis (aetiology, modification, lethality) a "protective influence" appears probable in female embryos, in blood group A (ABO system), in HLA antigens A9, A11, B35. An "embryotoxic influence" i.e., an increase in all the three influences is seen in male embryos, regional influences, in the B and AB blood groups, in HLA antigens B17, in primiparae, in multiple pregnancies, in older mothers and in cytomegalovirus infections. The Epstein-Barr virus seem to increase, in particular, CLP embryo lethality. The activity of the factors was rated by means of correlations between the cleft frequency in first-degree relatives and the frequency of the factor in six subgroups classified by the cleft defect subtypes, by means of interactions between two and more factors and by a study of the differences between male and female probands. A model of the threshold of CL and CP teratogenesis was proposed, a model in agreement with prenatal reciprocal equilibrium.


[Effects of low temperature exposure on dermal microvascular endothelial cells function].
To explore the damage effects and expression of vascular endothelial growth factor (VEGF) exposed with different low-temperatures on rat dermal microvascular endothelial cells (DMVECs). Primary DMVECs were obtained by discontinuous Percoll gradient centrifugation. The DMVECs were identified by phase contrast microscope and immunofluorescence studies for CD31 antigen. Applied 28 degrees C, 12 degrees C and 0 degrees C to interfere with rat DMVECs as cold-exposure model. The changes of cells morphology were observed under invert microscope. The membrane integrity was determined by lactate dehydrogenase (LDH) activity. RT-PCR was used to examine the expression of vascular endothelial growth factor mRNA in cells. The monolayer of cultured PMVECs displayed the shape of pavingstone. CD31 antigen and binding BSI results by fluorescence microscope identified the cultured cells were DMVECs. After 24 h cold exposure, the cell morphology of 0 degrees C group was shrunken, the other groups were "Fibroblast-like". The LDH activity (U/L) in the medium of 28 degrees C, 12 degrees C and 0 degrees C groups was 54.17 +/- 3.02, 64.66 +/- 3.03, 82.13 +/- 10.91 respectively, which was significantly higher than that of 37 degrees C group (12.23 +/- 3.0, P < 0.01). The VEGF mRNA expression level was up-regulated in 28 degrees C group and 12 degrees C group versus control group (P < 0.05), but unchanged in 0 degrees C group. The rat DMVECs injury severity are deteriorated with temperature decreasing, and VEGF might be involved in the regulation of membrane permeability in this period.


[The arcuate eminence of the petrous portion of the temporal bone. Descriptive features].
The superior semicircular canal is an important landmark in the middle cranial fossa approach to the temporal bone and the internal acoustic meatus. This mark is frequently located by its topographic association to the arcuate eminence, a bony prominence located on the anterior surface of petrous portion of the temporal bone. In recent studies, otolaryngologists described that the arcuate eminence lies more posterior and laterally than the superior semicircular canal and stated that some eminences corresponded to sulci of the temporal lobe, of which most were traces of the occipitotemporal sulcus. Our work examines by dissection the relationship of the arcuate eminence to the superior semicircular canal, demonstrating a third origin of the arcuate eminence: well pneumatized temporal bones in the region of the epi-tympanic recess.


Towards a tipping point in responding to change: rising costs, fewer options for Arctic and global societies.
Climate change incurs costs, but government adaptation budgets are limited. Beyond a certain point, individuals must bear the costs or adapt to new circumstances, creating political-economic tipping points that we explore in three examples. First, many Alaska Native villages are threatened by erosion, but relocation is expensive. To date, critically threatened villages have not yet been relocated, suggesting that we may already have reached a political-economic tipping point. Second, forest fires shape landscape and ecological characteristics in interior Alaska. Climate-driven changes in fire regime require increased fire-fighting resources to maintain current patterns of vegetation and land use, but these resources appear to be less and less available, indicating an approaching tipping point. Third, rapid sea level rise, for example from accelerated melting of the Greenland ice sheet, will create a choice between protection and abandonment for coastal regions throughout the world, a potential global tipping point comparable to those now faced by Arctic communities. The examples illustrate the basic idea that if costs of response increase more quickly than available resources, then society has fewer and fewer options as time passes.


Development of a Uterosacral Ligament Suspension Rat Model.
Pelvic organ prolapse (POP) is a common pelvic floor disorder (PFD) with the potential to significantly impact a woman's quality of life. Approximately 10%-20% of women undergo pelvic floor repair surgery to treat prolapse in the United States. PFD cases result in an overall $26.3 billion annual cost in the United States alone. This multifactorial condition has a negative impact on the quality of life and yet the treatment options have only dwindled in the recent past. One common surgical option is uterosacral ligament suspension (USLS), which is typically performed by affixing the vaginal vault to the uterosacral ligament in the pelvis. This repair has a lower incidence of complications compared to those with mesh augmentation, but is notable for a relatively high failure rate of up to 40%. Considering the lack of standard animal models to study pelvic floor dysfunction, there is an urgent clinical need for innovation in this field with a focus on developing cost-effective and accessible animal models. In this manuscript, we describe a rat model of USLS involving a complete hysterectomy followed by fixation of the remaining vaginal vault to the uterosacral ligament. The goal of this model is to mimic the procedure performed on women to be able to use the model to then investigate reparative strategies that improve the mechanical integrity of the ligament attachment. Importantly, we also describe the development of an in situ tensile testing procedure to characterize interface integrity at chosen time points following surgical intervention. Overall, this model will be a useful tool for future studies that investigate treatment options for POP repair via USLS.


[Two adolescent cases of pseudoseizure with mental retardation].
We experienced two cases of complicated pseudoseizure, whose diagnosis and treatment were based on Kretschmer's concept of "primitive hysteria". The first case was a 16-year-old boy who experienced a convulsion before a swimming class which he disliked. He was immediately brought to the emergency department of our hospital. Initially, he was treated as a true epileptic patient, and his convulsion continued for hours. We monitored the patient by video, which aided in making a precise diagnosis. He also showed a pain disorder. The second case was a 16-year-old girl who developed hyperventilation and convulsion during her graduation ceremony. The characteristics of her convulsion were similar to the first case. Four days after her admission to our hospital, we concluded that her symptoms were a part of primitive hysteria. After her discharge, she experienced some intermittent episodes of convulsion. There was also a possibility of sexual abuse from her father. Both patients had a family history of mental retardation and an unstable home life, as well as similar symptoms. Even in a modern general hospital, there is a lack of understanding about pseudoseizure, thus, medication may be unnecessary for undiagnosed patients. DSM-IV-TR as well as ICD-10 criteria do not mention anything about primitive hysteria. However, we recommend revitalization of this concept because it is a useful, appropriate, and necessary description of pseudoseizure with complications.


[Triage quickens the treatment of the most sick patients].
Systematic process triage is a relatively unknown concept in Denmark. Currently there are no national recommendations regarding triage models for use in the emergency department (ED). Four medium-sized EDs from different regions across the country cooperated in a joint venture to develop a new triage model, Danish Emergency Process Triage (DEPT). DEPT is inspired by the Swedish ADAPT system, but modified for a Danish context. This paper summarizes the cumulated experience with the new system.


How does blood oxygen level-dependent (BOLD) contrast correlate with oxygen partial pressure (pO2) inside tumors?
Blood oxygen level-dependent (BOLD) contrast-based functional MRI (fMRI) has been reported as a method to assess the evolution of tumor oxygenation after hyperoxic treatments, because of its sensitivity to changes in blood flow and deoxyhemoglobin content. However a number of questions remain: 1) In view of tumor heterogeneity, how good is the correlation between the MR parameters in gradient-echo imaging (signal intensity (SI) or effective transverse relaxation time (T(*)(2))) and local tumor oxygen partial pressure (pO(2))? 2) Is the magnitude of the change in SI or T(*)(2) a quantitative marker for variation in pO(2)? 3) Is initial T(*)(2) a good marker for initial pO(2)? To address these questions, murine tumors were imaged during respiratory challenges at 4.7 Tesla, using fiber-optic microprobes to simultaneously acquire tumor pO(2) and erythrocyte flux. The BOLD signal response (SI and T(*)(2)) was temporally correlated with changes in pO(2). However, the magnitude of the signal bore no absolute relation to pO(2) across tumors, i.e., a given change in SI corresponded to a 25 mmHg pO(2) change in one tumor, but to a 100 mmHg change in another. The initial T(*)(2) value did not reliably predict tumor oxygenation at the beginning of the experiment. In conclusion, the major advantages of the technique include noninvasiveness, high spatial resolution, and real-time detection of pO(2) fluctuations. Information afforded by the BOLD imaging technique is qualitative in nature and may be combined with other techniques capable of providing an absolute measure of pO(2).


Analysis of variation in cis-9, trans-11 conjugated linoleic acid (CLA) in milk fat of dairy cows.
Conjugated linoleic acid (CLA) is a fatty acid with numerous putative health benefits and is a natural component of ruminant-derived food products. An intermediate in rumen biohydrogenation is cis-9, trans-11 CLA, the major CLA isomer in milk fat. However, the major source of cis-9, trans-11 CLA in milk is endogenous synthesis by delta 9-desaturase conversion of trans-11 C18:1, another rumen biohydrogenation intermediate. The desaturase indices serve as a proxy for delta 9-desaturase activity and are calculated from the ratios of fatty acid pairs that represent product/substrate for this enzyme. This study analyzed individual animal variation in milk fat content of cis-9, trans-11 CLA and in desaturase indices in milk fat. Thirty lactating Holstein cows were allocated to one of three treatment groups: one received a standard total mixed ration, one received a diet that produced an elevated milk fat content of CLA, and a third treatment group was alternated between these diets at 3-wk intervals over the 12-wk study. There was a two- to threefold variation among individuals on the same diet for both milk fat content of CLA and desaturase indices in milk fat. This hierarchy was maintained to a large extent over the 12-wk study even in the variable treatment group that alternated between the two diets. Within the variable diet treatment, some animals consistently had a substantial response in milk fat content of CLA to dietary shifts, whereas other cows had little or no response. We conclude that while diet is a major determinant of the CLA content in milk fat, individual animal differences also have a substantial effect. The variation among individuals includes differences related to both rumen biohydrogenation and delta 9-desaturase activity in the mammary gland.


[Assessment of the preparation Carietest, Septodont].
The usefulness for staining of plaques on teeth was assessed using the preparation Carietest manufactured by Septodont. Carietest is a ready for use green fluid prepared, according to the producer, of blue sulphane and yellow tartrazin. It is applied painting the teeth in any area in which poor oral hygiene is suspected. The preparation was tried out using as reference standard Plakk-festö erythrosin tablets of Hungarian origin. The studied group comprised 25 patients. The range of staining of dental plaques by both preparations was determined using a simplified fuchsin index. The study showed a good suitability of the preparation Carietest for staining of bacterial dental plaques.


Pharmacological activation of soluble guanylate cyclase improves vascular graft function.
Ischaemia-reperfusion injury impairs the nitric oxide/soluble guanylate cyclase/cyclic guanosine monophosphate (cGMP) signalling pathway and leads to vascular dysfunction. We assessed the hypothesis that the soluble guanylate cyclase activator cinaciguat would protect the vascular graft against ischaemia-reperfusion injury. In the treatment groups, rats (n = 8/group) were pretreated with either intravenous saline or intravenous cinaciguat (10 mg/kg) 2 h before an aortic transplant. Aortic grafts were stored for 2 h in saline and transplanted into the abdominal aorta of the recipients. Two hours after the transplant, the grafts were harvested and mounted in an organ bath. Vascular function of the grafts was investigated in the organ bath. Terminal deoxynucleotidyl transferase dUTP nick end labelling, cluster of differentiation 31, caspase-3, endothelial nitric oxide synthase, cGMP, nitrotyrosine and vascular cell adhesion molecule 1 immunochemical reactions were also investigated. Pretreatment with cinaciguat significantly improved endothelium-dependent maximal relaxation 2 h after reperfusion compared with the saline group (maximal relaxation control: 96.5 ± 1%, saline: 40.4 ± 3% vs cinaciguat: 54.7 ± 2%; P < 0.05). Pretreatment with cinaciguat significantly reduced DNA fragmentation and nitro-oxidative stress; decreased the caspase-3 and vascular cell adhesion molecule 1 scores; and increased endothelial nitric oxide synthase, cGMP and cluster of differentiation 31 scores. Our results demonstrated that enhancement of cGMP signalling by pharmacological activation of the soluble guanylate cyclase activator cinaciguat might represent a beneficial therapy for treating endothelial dysfunction of arterial bypass graft during cardiac surgery.


The value of two-field pattern electroretinogram in routine clinical electrophysiologic practice.
To investigate the clinical use of the large-field pattern electroretinogram (PERG) as an adjunct test to the International-standard PERG in an unselected sequential cohort of patients referred for routine electrophysiologic assessment. Pattern electroretinograms to both 15° × 11° (International Society for Clinical Electrophysiology of Vision Standard) and 30° × 22° (large field) checkerboard field sizes were recorded in 277 consecutive electrophysiology patients, aged 10-79 years. Most patients had additional tests including full-field electroretinogram, electrooculogram, multifocal electroretinograms, or cortical visual evoked potential. Patient data were compared with data from 27 control subjects. Satisfactory 2-field PERG data were obtained in 91% (N = 253) of patients; data from 24 patients (9%) were excluded because of poor compliance (n = 17) or nystagmus (n = 7). Standard PERGs were consistent with macular dysfunction in 44% of cases; large-field PERG revealed macular dysfunction in an additional 8% of eyes and helped to distinguish between localized central, predominantly paracentral, and widespread macular dysfunction. The results were consistent with multifocal electroretinogram and/or imaging studies on the same patients. In some patients with optic nerve disease, the large-field PERG provided clearer evidence of normal macular function than the standard PERG. Routine use of the large-field PERG is a valuable complement to standard-field PERG testing in the evaluation and management of patients with different forms of macular or generalized retinal dysfunction and can be useful in patients with optic nerve disease.


Removing intensity effects and identifying significant genes for Affymetrix arrays in macrophage migration inhibitory factor-suppressed neuroblastoma cells.
A semilinear in-slide model is introduced to remove the intensity effect in the scanning process. It is demonstrated that the intensity effect can be estimated accurately and removed effectively. This normalization step is vital for Affymetrix arrays to reveal relevant biological results when comparing gene expression in multiple arrays. The normalized expression ratios are analyzed further by a modified two-sample t test along with a sieved permutation scheme for computing P values. The improved specificity and sensitivity are demonstrated by using a study on the impact of macrophage migration inhibitory factor (MIF) reduction in neuroblastoma cells. With semilinear in-slide model analysis, expression of 166 genes was altered with a P value no greater than 0.001. Among those genes, 44 were altered >2-fold. MIF-regulated genes associated with tumor development including IL-8 and C-met, which are overexpressed in many tumors, were down-regulated in MIF-reduced cells. On the other hand, some tumor-suppressor genes such as EPHB6, visinin-like protein 1 (VSNL-1), and BLU were up-regulated in MIF-reduced cells. In addition, we demonstrated that down-regulation of MIF expression could result in a reduction in cell proliferation and tumor growth in vitro and in vivo. Our data not only demonstrate that targeting MIF expression is a promising therapeutic strategy in human neuroblastoma therapy but also indicate the MIF target genes for additional study.


Regulation of the corticosteroid signalling system in rainbow trout HPI axis during confinement stress.
This study aims to shed light on corticosteroid regulation of stress in teleost fish with focus on the corticosteroid signalling system. The role of the mineralocorticoid-like hormone 11-deoxycorticosterone (DOC) in fish is still enigmatic, as is the function of the mineralocorticoid receptor, MR. Low plasma DOC levels and ubiquitous tissue distribution of MR question the physiological relevance of the mineralocorticoid-axis. Furthermore, the particular purpose of each of the three corticosteroid receptors in fish, the glucocorticoid receptors, GR1 and GR2, and the MR, is still largely unknown. Therefore we investigate the regulation of cortisol and DOC in plasma and mRNA levels of MR, GR1 and GR2 in the HPI-axis tissues (hypothalamus, pituitary and interrenal gland) during a detailed confinement stress time-course. Here we show a sustained up-regulation of plasma DOC levels during a confinement stress time-course. However, the low DOC levels compared to cortisol measured in the plasma do not favour an activity of DOC through MR receptors. Furthermore, we show differential contribution of the CRs in regulation and control of HPI axis activity following confinement stress. Judged by the variation of mRNA levels negative feedback regulation of cortisol release occurs on the level of the pituitary via MR and on the level of the interrenal gland via GR2. Finally, asa significant effect of confinement stress on CR expressions was observed in the pituitary gland, we completed this experiment by demonstrating that corticosteroid receptors (GR1, GR2 and MR) are co-expressed in the ACTH cells located in the adenohypophysis. Overall, these data suggest the involvement of these receptors in the regulation of the HPI axis activity by cortisol.


Estimation of Photorespiration Based on the Initial Rate of Postillumination CO(2) Release: I. A Nonsteady State Model for Measurement of CO(2) Exchange Transients.
Although open systems have been used for the study of transients in leaf CO(2) exchange such as the postillumination burst, these systems frequently do not permit reliable estimates of transient rates due to their nonsteady state nature. A nonsteady state mathematical approach is described which predicts changes in CO(2) concentration in the leaf chamber and infrared gas analyzer measuring cell as a function of leaf CO(2) exchange rate in Nicotiana tabacum vars John Williams Broadleaf and Havana Seed. With the aid of a computer, a numerical formula simulates the mixing and dilution which occurs as CO(2) passes through the finite volume of the measuring cell of the analyzer. The method is presented with special relevance to photorespiration as manifested by the postillumination burst of CO(2). The latter is suggested to decline with the first order kinetics following darkening of a C(3) leaf. This approach provides a basis for reliable estimation of the initial and, hence, maximal rate of CO(2) evolution during the postillumination burst under a variety of environmental conditions.


Frequency of serological non-responders and false-negative RT-PCR results in SARS-CoV-2 testing: a population-based study.
Objectives The sensitivity of molecular and serological methods for COVID-19 testing in an epidemiological setting is not well described. The aim of the study was to determine the frequency of negative RT-PCR results at first clinical presentation as well as negative serological results after a follow-up of at least 3 weeks. Methods Among all patients seen for suspected COVID-19 in Liechtenstein (n=1921), we included initially RT-PCR positive index patients (n=85) as well as initially RT-PCR negative (n=66) for follow-up with SARS-CoV-2 antibody testing. Antibodies were detected with seven different commercially available immunoassays. Frequencies of negative RT-PCR and serology results in individuals with COVID-19 were determined and compared to those observed in a validation cohort of Swiss patients (n=211). Results Among COVID-19 patients in Liechtenstein, false-negative RT-PCR at initial presentation was seen in 18% (12/66), whereas negative serology in COVID-19 patients was 4% (3/85). The validation cohort showed similar frequencies: 2/66 (3%) for negative serology, and 16/155 (10%) for false negative RT-PCR. COVID-19 patients with negative follow-up serology tended to have a longer disease duration (p=0.05) and more clinical symptoms than other patients with COVID-19 (p<0.05). The antibody titer from quantitative immunoassays was positively associated with the number of disease symptoms and disease duration (p<0.001). Conclusions RT-PCR at initial presentation in patients with suspected COVID-19 can miss infected patients. Antibody titers of SARS-CoV-2 assays are linked to the number of disease symptoms and the duration of disease. One in 25 patients with RT-PCR-positive COVID-19 does not develop antibodies detectable with frequently employed and commercially available immunoassays.


Selected clinical features of the head and neck in women with Turner syndrome and the 45,X/46,XY karyotype.
A 45,X/46,XY karyotype in women with Turner syndrome (TS) is very rare. The presence of a Y chromosome in the karyotype causes phenotypic differences and increased risk for neoplastic disease, compared to TS-women with other karyotypes. Our study addresses an issue: non-genital phenotypic differences between TS-patients with a Y-chromosome of their karyotype and TS-women without it. Results from patient history/physical examinations of the head and neck of eight TS-women and the 45,X/46,XY karyotype were compared with those observed in 164 TS-women and 30 controls. The heights of TS-groups: 142.5 ± 7.2 and 144.9 ± 7.2 cm were lower than controls (165.2 ± 6.6 cm). Participants were examined from 1995 to 2014. Among 28 study parameters, 15 were more frequently observed in TS women with the 45,X/46,XY karyotype compared to controls. Only abnormalities in the oral cavity and a history of childhood lymphoedema, differed significantly in the TS groups. With respect to the head and neck, the patient history and physical examination results of TS-women and the 45,X/46,XY karyotype and TS and other karyotypes revealed similar differences compared to controls. Compared to others TS patients, 45,X/46,XY individuals might more frequently have oral cavity soft tissue abnormalities and more rarely a history of childhood lymphoedema. (Endokrynol Pol 2017; 68 (1): 47-52).


Identification of a hamster sperm 26-kilodalton dehydrogenase/reductase that is exclusively localized to the mitochondria of the flagellum.
Sperm mitochondria undergo remodeling during posttesticular maturation that includes extensive disulfide cross-linking of proteins of the outer membrane to form the insoluble mitochondrial capsule. The relationship of these changes to mitochondrial function in mature gametes is unclear. The phospholipid hydroperoxide glutathione peroxidase (GPX4; also termed PHGPx) represents a major disulfide bond-stabilized protein of the mitochondrial capsule, and it is readily released by disulfide-reducing agents. However, in addition to GPX4, we detected a second major protein of 26 kDa (MP26) that was eluted from purified hamster sperm tails by the disulfide-reducing agent dithiothreitol. The objectives of the present study were to identify and characterize MP26 and to explore its potential role in mitochondrial function. Proteomic analysis of MP26 by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) identified 14 peptides with sequence identity to a member of the short-chain dehydrogenase/reductase superfamily termed P26h, which was implicated previously in hamster sperm-zona binding, and with high sequence similarity to mouse lung carbonyl reductase. Indirect immunofluorescence localized MP26 to the midpiece, and two-dimensional PAGE and immunoblot analysis identified a single MP26 isoform of pI 9.0. Immunoblot analyses of cauda epididymal fluid and of purified sperm plasma membranes and mitochondria revealed the exclusive localization of MP26 to the mitochondrial fraction. These data indicate that MP26 does not function in zona binding; instead, like GPX4, it may be associated with the mitochondrial capsule and play an important role in sperm mitochondrial function.


Accessing Active Inference Theory through Its Implicit and Deliberative Practice in Human Organizations.
Active inference theory (AIT) is a corollary of the free-energy principle, which formalizes cognition of living system's autopoietic organization. AIT comprises specialist terminology and mathematics used in theoretical neurobiology. Yet, active inference is common practice in human organizations, such as private companies, public institutions, and not-for-profits. Active inference encompasses three interrelated types of actions, which are carried out to minimize uncertainty about how organizations will survive. The three types of action are updating work beliefs, shifting work attention, and/or changing how work is performed. Accordingly, an alternative starting point for grasping active inference, rather than trying to understand AIT specialist terminology and mathematics, is to reflect upon lived experience. In other words, grasping active inference through autoethnographic research. In this short communication paper, accessing AIT through autoethnography is explained in terms of active inference in existing organizational practice (implicit active inference), new organizational methodologies that are informed by AIT (deliberative active inference), and combining implicit and deliberative active inference. In addition, these autoethnographic options for grasping AIT are related to generative learning.


VARAdb: a comprehensive variation annotation database for human.
With the study of human diseases and biological processes increasing, a large number of non-coding variants have been identified and facilitated. The rapid accumulation of genetic and epigenomic information has resulted in an urgent need to collect and process data to explore the regulation of non-coding variants. Here, we developed a comprehensive variation annotation database for human (VARAdb, http://www.licpathway.net/VARAdb/), which specifically considers non-coding variants. VARAdb provides annotation information for 577,283,813 variations and novel variants, prioritizes variations based on scores using nine annotation categories, and supports pathway downstream analysis. Importantly, VARAdb integrates a large amount of genetic and epigenomic data into five annotation sections, which include 'Variation information', 'Regulatory information', 'Related genes', 'Chromatin accessibility' and 'Chromatin interaction'. The detailed annotation information consists of motif changes, risk SNPs, LD SNPs, eQTLs, clinical variant-drug-gene pairs, sequence conservation, somatic mutations, enhancers, super enhancers, promoters, transcription factors, chromatin states, histone modifications, chromatin accessibility regions and chromatin interactions. This database is a user-friendly interface to query, browse and visualize variations and related annotation information. VARAdb is a useful resource for selecting potential functional variations and interpreting their effects on human diseases and biological processes.


Fiber-based source of 500 kW mid-infrared solitons.
Fiber-based sources delivering high-energy few-cycle pulses at high repetition rates are currently being developed in the near-infrared spectral range, thanks to the wide availability of telecommunication-grade optical fibers and components. Similar sources in the middle-wave infrared (mid-IR) spectral domain, however, are scarce, although such sources are of high interest for applications such as high-precision frequency metrology and molecular spectroscopy or as a seed source to reach further into the mid-IR via coherent nonlinear processes. Here we report on the design of a fiber-based source of 50-nJ energy 90 fs duration pulses up to 2950 nm, corresponding to 500 kW peak power. To obtain this level of peak power we exploit multi-solitonic fission and soliton self-frequency shift in large mode area fibers excited by picosecond pulses emitted at 2 µm from a megahertz repetition rate fiber laser. We leverage mature silica-based fiber technology up to 2.4 µm and restrict the use of fluoride fiber to the very last frequency-shifting stage. The level of instantaneous power and ultra-short duration achieved in this Letter pave the way to all-fiber format generation of an ultra-broadband coherent continuum in the mid-IR with profound implications for applications such as high-resolution molecular spectroscopy and imaging.


Fire management effects on ruminal digestibility and in vitro methane emissions of subtropical rangeland plant species.
Prescribed fire is a common management practice used to manipulate rangeland plant productivity and composition. Although the nutritive value of most herbaceous plant species is considered poor for grazing animals, native rangelands in Florida are an important source of forage for livestock, especially during the winter months, when the productivity of cultivated perennial warm-season pastures is limited. This study evaluated the effects of prescribed fire on methanogenic potential and nutritive value of selected native rangeland plant species. Treatments were a 3 × 2 factorial arrangement of plant species (creeping bluestem [Schizachyrium scoparium var. stoloniferum {Nash} Wipff], wiregrass [Aristida stricta {Michx.}], or saw palmetto [Serenoa repens {W. Bartram} Small]) and prescribed fire management [2 yr after burning (control) vs. 1 yr after burning (burned)] distributed in a randomized complete block design with four replicates. Samples were analyzed for crude protein (CP), neutral detergent undigestible fiber (NDF), in vitro methane production, and in situ ruminal disappearance. Prescribed fire generally increased forage CP and DM effective degradability relative to control; however, no effect was observed on saw palmetto. Wiregrass had the least CP concentration in both burned (8.5%) and control (2.3%). In burned treatments, creeping bluestem and palmetto had greater DM effective degradability (62% and 58%) than wiregrass (53%). Fire increased in vitro gas production by 60 (creeping bluestem) to 90% (wiregrass) relative to control treatments. No effect of fire on methane production was observed for any of the plant species evaluated in this study. Creeping bluestem had the greatest methane production (12.5 mg/g DM), followed by wiregrass (5.3 mg/g DM) and saw palmetto (1.4 mg/g DM). Methane:DM effective degradability decreased in the following order: creeping bluestem ≥ wiregrass > saw palmetto. Data indicated prescribed fire was an effective tool to increase creeping bluestem and wiregrass nutritive value but no effect was observed on saw palmetto. Cattle grazing grass-dominated rangelands will likely emit more gas and methane than shrub or tree-dominated ecosystems; however, the greater forage nutritive value and subsequent positive impacts on animal production are expected to offset a substantial fraction of enteric methane emissions.


Infrared suppression by hybrid EUV multilayer--IR etalon structures.
We have developed a multilayer mirror for extreme UV (EUV) radiation (13.5 nm), which has near-zero reflectance for IR line radiation (10.6 μm). The EUV reflecting multilayer is based on alternating B4C and Si layers. Substantial transparency of these materials with respect to the IR radiation allowed the integration of the multilayer coating in a resonant quarter-wave structure for 10.6 μm. Samples were manufactured using magnetron sputtering deposition technique and demonstrated suppression of the IR radiation by up to 3 orders of magnitude. The EUV peak reflectance amounts 45% at 13.5 nm, with a bandwidth at FWHM being 0.284 nm. Therefore such a mirror could replace conventional multilayer mirrors to suppress undesired spectral components in monochromatic imaging applications, including EUV photolithography.


A patient-specific numerical modeling of the spontaneous coronary artery dissection in relation to atherosclerosis.
The spontaneous coronary artery dissection (SCAD) is a clinical complication of angioplasty leading to an initiation of a tear/crack in the intima layer of the artery. The crack can propagate to the interface of the intima-media layer following by intramural hematoma. The relation between the SCAD and atherosclerosis is a controversial issue, as some studies stated no correlation between them while others showed that a crack can initiate in the intima but cannot propagate into the atrophied media layer. To investigate the relation between the intraluminal crack propagation in the atherosclerotic artery and SCAD, this study numerically investigated the initiation and propagation of a crack in the intraluminal and radial locations of the healthy and atherosclerotic human coronary arterial walls. The energy release rate, namely J-integral, is computed as a numerical derivative of the strain energy with respect to a crack extension using a user-defined virtual crack method (VCE) of extended finite element method (XFEM). Experimental measurements were carried out to calculate the elasto-plastic mechanical properties of the healthy and atherosclerotic human coronary arteries. The experimental data were then assigned to our own established patient-specific FE model of the coronary artery. Cracks were sketched in the intraluminal and radial locations of the arterial wall and allowed to propagate to the virtual interface of the intima-media to form a false lumen. The results revealed a higher stress at the crack tip of the healthy arterial wall compared to the atherosclerotic one. Lower crack tip opening displacement (CTOD) and crack tip opening angle (CTOA) were observed in the intraluminal crack of the atherosclerotic artery. J-integral of the atherosclerotic arterial wall was also found to be higher than the healthy one at the intraluminal crack. The results revealed that although a crack can initiate in the intraluminal of an atherosclerotic artery, it cannot propagate into the media layer due to a relatively higher rate of the strain energy release in the atherosclerotic arterial wall compared to the healthy one.


An Empirical and Subjective Model of Upper Extremity Fatigue Under Hypogravity.
In the context of extra-terrestrial missions, the effects of hypogravity (0 < G < 1) on the human body can reduce the well-being of the crew, cause musculoskeletal problems and affect their ability to perform tasks, especially during long-term missions. To date, studies of the effects of hypogravity on human movement are limited to experiments on the lower limbs. Here, we extend the knowledge base to the upper limbs, by conducting experiments to evaluate the effect of hypogravity on upper limb physical fatigue and mental workload in participants. Our hypothesis was that hypogravity would both increase participant productivity, by reducing overall physical fatigue expressed in Endurance Time, and reduce mental workload. Task Intensity-Endurance time curves are developed especially in seated positions, while performing static, dynamic, repetitive tasks. This experiment involved 32 healthy participants without chronic problems of the musculoskeletal system aged 33.59 ± 8.16 years. Using the collected data, fatigue models were constructed for tasks of varying Intensity. In addition, all participants completed the NASA - Task Load Index subjective mental workload assessment, which revealed the level of subjective workload when executing different tasks. We found two trends in the empirical fatigue models associated with the difference between the strength capabilities of males and females. The first is a significant positive (p = 0.002) relation between Endurance time and gravity level (⅙ G Moon, ⅓ G Mars, 1G) with negative coefficient for males and females for a static task. And there is marginal relation (p < 0.1) between overall mental workload and gravity level with a positive coefficient for males and females for the same task. The same trend was observed for dynamic and repetitive tasks. We concluded that the Task Intensity-Endurance Time model, adapted to hypogravity in combination with subjective mental assessment, is useful to human fatigue investigation. The combination of these methods used for ergonomic analysis and digital human modeling, could improve worker productivity. Finally, this study may help prepare astronauts for long-term missions on the Moon and Mars and improve our understanding of how we can prevent musculoskeletal disorders caused by hazardous manual handling under such extreme environments.


Venoarterial Extracorporeal Membrane Oxygenation Support as a Bridge to Heart Transplant: Report of 3 Cases.
Heart transplant is the only definitive treatment of end-stage heart failure. Venoarterial extracorporeal membrane oxygenation may be used as a bridge to heart transplant. Among 31 patients who underwent heart transplant between January 2014 and June 2016, we present our experiences with 3 patients who received venoarterial extracorporeal support as a bridge to heart transplant. The first patient was a 51-year-old male with ischemic dilated cardiomyopathy. Transplant was performed after 6 days of extracorporeal support, and the patient was discharged and alive at follow-up. Patient 2 was a 12-yearold girl with dilated cardiomyopathy who presented with cardiac arrest. Extracorporeal support was initiated during cardiopulmonary resuscitation. She had full neurologic recovery and remained on the wait list. She received a transplant 22 days after resuscitation. She survived and was alive at day 220 posttransplant. The third patient was a 50-year-old male with ischemic dilated cardiomyopathy requiring venoarterial extracorporeal support. Percutaneous balloon atrial septostomy was performed for left ventricle venting. He underwent transplant on day 28 after intensive care unit admission. He died 29 days after release from the hospital. Regarding patients on heart transplant wait lists who are worsening despite optimal medical therapy, venoarterial extracorporeal membrane oxygenation support is a safe and viable last resort.


Study of Sedative Tea Phytocomplex within the Framework of Studies Aimed at Creation of a Rectal Dosage Form with Antihistaminic Effect.
We designed a new complex drug with antiallergic effect containing, in addition to the main component loratadine, a phytocomplex for an extra therapeutic effect. A collection of plants with sedative activity is chosen and the optimal agent for extraction of bioactive compounds (40% ethanol) and optimal degree of plant fragmentation are determined. Chemical composition of the sedative tea is evaluated by reverse phase HPLC. The marker components of the species are detected: xanthohumol and isoxanthohumol - Humulus lupulus cone components, Mentha piperita rosmarinic acid, and scutellareine, Menyanthes trifolia element - quercetin-3-rutinoside, and caffeic acid. Standardization of the species by the absolute graduation method in conversion to quercetin-3-rutinoside is suggested.


Machine learning predictive model for severe COVID-19.
To develop a modified predictive model for severe COVID-19 in people infected with Sars-Cov-2. We developed the predictive model for severe patients of COVID-19 based on the clinical date from the Tumor Center of Union Hospital affiliated with Tongji Medical College, China. A total of 151 cases from Jan. 26 to Mar. 20, 2020, were included. Then we followed 5 steps to predict and evaluate the model: data preprocessing, data splitting, feature selection, model building, prevention of overfitting, and Evaluation, and combined with artificial neural network algorithms. We processed the results in the 5 steps. In feature selection, ALB showed a strong negative correlation (r = 0.771, P < 0.001) whereas GLB (r = 0.661, P < 0.001) and BUN (r = 0.714, P < 0.001) showed a strong positive correlation with severity of COVID-19. TensorFlow was subsequently applied to develop a neural network model. The model achieved good prediction performance, with an area under the curve value of 0.953(0.889-0.982). Our results showed its outstanding performance in prediction. GLB and BUN may be two risk factors for severe COVID-19. Our findings could be of great benefit in the future treatment of patients with COVID-19 and will help to improve the quality of care in the long term. This model has great significance to rationalize early clinical interventions and improve the cure rate.


Coronary artery disease is associated with Alzheimer disease neuropathology in APOE4 carriers.
To examine the associations between postmortem Alzheimer disease (AD) neuropathology and autopsy-verified cardiovascular disease. The authors examined 99 subjects (mean age at death = 87.6; SD = 8.7) from the Mount Sinai School of Medicine Department of Psychiatry Brain Bank who were devoid of cerebrovascular disease-associated lesions or of non-AD-related neuropathology. Density of neuritic plaques (NPs) and neurofibrillary tangles (NFTs) as well as coronary artery and aortic atherosclerosis, left ventricular wall thickness, and heart weight were measured. Partial correlations were used to assess the associations of the four cardiovascular variables with NPs and NFTs in the hippocampus, entorhinal cortex, and multiple regions of the cerebral cortex after controlling for age at death, sex, dementia severity, body mass index, and ApoE genotype. These analyses were also repeated separately for ApoE4 carriers and noncarriers. The extent of coronary artery disease and to a lesser extent atherosclerosis were significantly associated with the density of cardinal neuropathologic lesions of AD in this autopsy sample (significant correlations between 0.22 and 0.29). These associations were more pronounced for the ApoE4 allele carriers (n = 42; significant correlations between 0.34 and 0.47). The degree of coronary artery disease is independently associated with the cardinal neuropathological lesions of Alzheimer disease. These associations are primarily attributable to individuals with the ApoE4 allele.


Infusion of nursing informatics: from Outside In.
Knowledge about informatics is essential for all nurses. Many faculties know and employ parts of nursing informatics in their courses and curriculum, however the infusion of technology enhanced learning and informatics into the curriculum is expensive, time consuming and takes expertise. To address these issues a 5-institution Nursing Collaborative designed a five-year plan that has each of the institutions responsible to provide informatics teaching, learning and resources to faculty and students at all five institutions - moving learning from Outside In.


DVT prophylaxis in the perioperative setting.
In last month's BJPB, Agnes Arnold discussed the pathophysiology of deep vein thrombosis and suggested ways that the risk could be reduced, particularly with respect to DVTs resulting from endothelial damage in surgery. In this, the second of a two-part series, Agnes continues by looking at hypercoagulability of blood and venous stasis and the impact of these conditions on the risk of DVT.


Association of a microsomal triglyceride transfer protein gene polymorphism with blood pressure in Japanese women.
Genetic variants of the microsomal triglyceride transfer protein (MTP) have been associated with the serum concentration of low density lipoprotein-cholesterol, predisposition to coronary heart disease, or longevity. The relation of a -493Gright curved arrow T polymorphism in the promoter of MTP to blood pressure was examined in a population-based study. The subjects (1124 men, 1108 women) were aged 40-79 years and were randomly recruited to a population-based prospective cohort study of aging and age-related diseases in Japan. Blood pressure was measured at least twice with subjects in the sitting position. The serum lipid profile was determined after the subjects had fasted overnight. The -493Gright curved arrow T genotype of MTP was determined with a fluorescence-based allele-specific DNA primer assay system. There was no difference in the serum concentrations of total cholesterol, high density lipoprotein-cholesterol, low density lipoprotein-cholesterol, or triglycerides among MTP genotypes for men or for women. Systolic or diastolic blood pressure was not related to the -493Gright curved arrow T polymorphism in men. For women, however, systolic and diastolic blood pressures were significantly related to MTP genotype, with the T allele of the polymorphism being associated with low blood pressure. The relation between MTP genotype and the prevalence of hypertension was almost significant (P=0.055) for all women. Although MTP genotype was not associated with the prevalence of hypertension in premenopausal women, the relation between these parameters was significant (P=0.040) in postmenopausal women, with the TT genotype protecting against this condition. These results suggest that MTP genotype is a determinant of blood pressure in Japanese women.


Echocardiographic abnormalities and predictors of mortality in hospitalized COVID-19 patients: the ECHOVID-19 study.
The present study had two aims: (i) compare echocardiographic parameters in COVID-19 patients with matched controls and (2) assess the prognostic value of measures of left (LV) and right ventricular (RV) function in relation to COVID-19 related death. In this prospective multicentre cohort study, 214 consecutive hospitalized COVID-19 patients underwent an echocardiographic examination (by pre-determined research protocol). All participants were successfully matched 1:1 with controls from the general population on age, sex, and hypertension. Mean age of the study sample was 69 years, and 55% were male participants. LV and RV systolic function was significantly reduced in COVID-19 cases as assessed by global longitudinal strain (GLS) (16.4% ± 4.3 vs. 18.5% ± 3.0, P < 0.001), tricuspid annular plane systolic excursion (TAPSE) (2.0 ± 0.4 vs. 2.6 ± 0.5, P < 0.001), and RV strain (19.8 ± 5.9 vs. 24.2 ± 6.5, P = 0.004). All parameters remained significantly reduced after adjusting for important cardiac risk factors. During follow-up (median: 40 days), 25 COVID-19 cases died. In multivariable Cox regression reduced TAPSE [hazard ratio (HR) = 1.18, 95% confidence interval (CI) [1.07-1.31], P = 0.002, per 1 mm decrease], RV strain (HR = 1.64, 95%CI[1.02;2.66], P = 0.043, per 1% decrease) and GLS (HR = 1.20, 95%CI[1.07-1.35], P = 0.002, per 1% decrease) were significantly associated with COVID-19-related death. TAPSE and GLS remained significantly associated with the outcome after restricting the analysis to patients without prevalent heart disease. RV and LV function are significantly impaired in hospitalized COVID-19 patients compared with matched controls. Furthermore, reduced TAPSE and GLS are independently associated with COVID-19-related death.


[Recurrent lower gastrointestinal bleeding due to a gastrointestinal stromal tumor (GIST) of the small intestine].
Gastrointestinal bleeding is a common cause of hospitalisation. In most patients, the origin and location of bleeding can be established in routine endoscopic and radiological examinations. The undiagnosed cases of so-called gastrointestinal bleeding of unknown origin require more sophisticated diagnostic methods. We present a case of 35-year old man with recurrent gastrointestinal bleeding due to a gastrointestinal stromal tumor of the jejunum that was revealed in the visceral angiography.