[Competence centre for victims of violence as part of the public health services--a consecutive case series of the "Schutzambulanz Fulda"].
Health-care services often fail to address violence sufficiently and to provide adequate forensic medical examination and documentation, especially in cases of intimate partner violence and violence against people in need of care. The "Schutzambulanz Fulda", hosted by the public health department of Fulda county, aims to close this gap in the east Hessian region. We have evaluated the first year of service. Between November 26(th) 2010 and November 26(th) 2011 a total of 154 persons called for support. Data concerning personal characteristics of the victims and the suspected perpetrators, the time and place of the incidents, and their impacts were analysed. 76.6% of th subjects were female, 23.4% male and 18.8% were less than 18 years old. In 60 cases a documentation of the injuries suitable for use in court was made. The majority of clients were women who reported having been assaulted by their partner or ex-partner. Community violence was less frequently stated. Contact to people in need of care suffering from violence was very rare. Currently, the "Schutzambulanz" as part of a public health department meets the goals to some degree. A comparative evaluation could help to identify the benefits and disadvantages of various trusteeships within the health-care system.


[Phospholipase D activity in maize seedling roots under salt stress and presowing treatment by preparations of adaptogenic action].
The effect of different salinity level and synthetic compounds treatment on phospholipase D activity in the root tissue of maize seedlings and the content of phosphatidic acid in plasmatic membrane has been investigated. The salt exposition to 0.05 M NaCl raised the activity of phospholipase D and the content of phosphatidic acid in the plasma membrane. The salt exposition to 0.1 M NaCl lowered the activity of phospholipase D, but raised the content of phosphatidic acid in the plasma membrane. The synthetic compounds treatment increased the activity of phospholipase D. It was shown that methyure treatment decreased the content of phosphatidic acid in the plasma membrane. The ivin treatment had the opposite effect.


Application of the stepwise focusing method to optimize the cost-effectiveness of genome-wide association studies with limited research budgets for genotyping and phenotyping.
The recent cataloguing of a large number of SNPs enables us to perform genome-wide association studies for detecting common genetic variants associated with disease. Such studies, however, generally have limited research budgets for genotyping and phenotyping. It is therefore necessary to optimize the study design by determining the most cost-effective numbers of SNPs and individuals to analyze. In this report we applied the stepwise focusing method, with two-stage design, developed by Satagopan et al. (2002) and Saito & Kamatani (2002), to optimize the cost-effectiveness of a genome-wide direct association study using a transmission/disequilibrium test (TDT). The stepwise focusing method consists of two steps: a large number of SNPs are examined in the first focusing step, and then all the SNPs showing a significant P-value are tested again using a larger set of individuals in the second focusing step. In the framework of optimization, the numbers of SNPs and families and the significance levels in the first and second steps were regarded as variables to be considered. Our results showed that the stepwise focusing method achieves a distinct gain of power compared to a conventional method with the same research budget.


Effect of oral cholecalciferol supplementation at physiological and supraphysiological doses in naturally vitamin D3-deficient subterranean damara mole rats (Cryptomys damarensis).
The damara mole rat, Cryptomys damarensis, is a strictly subterranean dwelling herbivorous rodent that in its natural habitat has no access to any obvious source of cholecalciferol (D3). We examined the effects of D3 supplementation, at physiological and supraphysiological doses, on calcium metabolism, plasma concentrations of calcium and alkaline phosphatase (ALP) and D3 metabolites. Animals not receiving a D3 supplement maintained normal plasma calcium concentrations. In addition, they exhibited a high apparent fractional mineral absorption efficiency (91%) and maintained a positive mineral flux. The serum concentration of 25-(OH)D3 was undetectable (less than 5 nmol/l) and that of 1,25-(OH)2D3 was 41 +/- 10 pmol/l. Supplementation at a physiological dose of D3 resulted in increased plasma concentrations of D3 metabolites, food intake, apparent fractional absorption efficiency and apparent fractional retention efficiency. Despite the 1.8-fold increase in food intake, body mass remained constant suggesting that the enhanced energy intake was dissipated in catabolic processes. Plasma calcium and ALP concentrations were not significantly altered with physiological doses of D3. The group given supraphysiological doses of D3 exhibited hypercalcaemia, increased creatinine concentrations and markedly increased ALP levels. These data indicate that a pathological response to D3 intoxication occurred and that hepatic and renal excretory functions were impaired. It appears, therefore, that these animals function optimally at the low concentrations of D3 metabolites found naturally. Supplementation at both physiological and supraphysiological doses of D3 may disadvantage the damara mole rat.


Glial cells in the developing and adult olfactory lobe of the moth Manduca sexta.
The antennal lobe of the moth contains several classes of glial cells that are likely to play functional roles in both the developing and mature lobe. In this study, confocal and electron microscopy were used to examine in detail the morphology of two classes of glial cells, those associated with olfactory receptor axons as they course to their targets in the lobe and those that form borders around the synaptic neuropil of the olfactory glomeruli. The former, the nerve-layer glia, have long processes with multiple expansions that enwrap axon fascicles; the latter, the neuropil glia, constitute two subgroups: complex glia with large cell bodies and branching, vellate arbors; and simple glia, with multiple, mostly unbranched processes with many lamellate expansions along their lengths. The processes of complex glia appear to be closely associated with axon fascicles as they enter the glomeruli, while those of the simple glia surround the glomeruli as part of a multi-lamellar glial envelope, their processes rarely invading the synaptic neuropil of the body of the glomerulus. The full morphological development of antennal-lobe glial cells requires more than two-thirds of metamorphic development. During this period, cells that began as cuboidal or spindle-shaped cells that were extensively dye-coupled to one another gradually assume their adult form and, at least under nonstimulated conditions, greatly reduce their coupling. These changes are only weakly dependent on the presence of olfactory receptor axons. Glial processes are somewhat shorter and less branched in the absence of these axons, but basic structure and degree of dye-coupling are unchanged.


Familial aggregation of nonhematological malignancies in relatives of patients with hematological neoplasms.
Familial aggregation of nonhematological malignant disorders (NHMD) was compared in 189 families of patients with hematological neoplasms (HN) with a control group of 36 families of patients with benign hematological disorders and a second group of 33 families of patients with diabetes mellitus. A self-administered questionnaire was used requesting from each family a full list of first- and second-degree relatives, their vital status, current age or age at death, and a list of their chronic diseases, including all malignant disorders. There was no evidence of a significantly increased tendency for developing NHMD among relatives of patients with HN as compared to controls (adjusted odds ratio of 0.88; 95% confidence interval 0.61-1.27). Moreover, in the HN group, no significant difference in the frequency of NHMD was found between the families with and without familial aggregation of HN. Based on the present analysis and our previous observations on familial aggregation of HN, we conclude that the increased aggregation of malignant disorders among relatives of patients with HN is unique to the hematopoietic system and might result from a genetic predisposition to HN in these families.


Surface marker analysis of acute myeloblastic leukemia: identification of differentiation-associated phenotypes.
A series of monoclonal antibodies reactive with normal myeloid cells at different stages of differentiation (anti-MY4, -MY7, -MY8, -Mo1, -Ia) were used to characterize the leukemic cells of 70 patients with acute myeloblastic leukemia (AML). Sixty-two of the leukemias expressed a phenotype corresponding to a recognizable immature normal myeloid cell. These 62 cases could be divided into 4 phenotype groups, corresponding approximately to the normal CFU-C (group I, 21%), myeloblast (group II, 26%), promyelocyte (group III, 8%), and promonocyte (group IV, 45%). Morphological subtyping of these leukemias tended to agree with the immunologic phenotype, particularly with more "differentiated" morphological subtypes, such as acute monocytic leukemia or acute promyelocytic leukemia. However, each phenotype group contained more than one morphological type of AML, indicating that the level of differentiation of the surface membrane of AML cells may not always be concordant with morphology. The phenotype groups were also analyzed with respect to cytochemical staining patterns, age, the presence of Auer rods, and complete remission rates. Statistically significant differences among the groups were noted in the distribution of myeloperoxidase staining, nonspecific esterase staining, and Auer rods. The complete remission rates varied from 60% (groups III and IV) to 88% (group II). These results suggest that surface marker analysis in AML may be used as a highly reproducible classification system that will provide additional information about the leukemic cells in conjunction with morphological analysis.


Variation in resistance to hydrostatic pressure among strains of food-borne pathogens.
Among food-borne pathogens, some strains could be resistant to hydrostatic pressure treatment. This information is necessary to establish processing parameters to ensure safety of pressure-pasteurized foods (N. Kalchayanand, A. Sikes, C. P. Dunne, and B. Ray, J. Food Prot. 61:425-431, 1998). We studied variation in pressure resistance among strains of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella species at two temperatures of pressurization. Early-stationary-phase cells in 1% peptone solution were pressurized at 345 MPa either for 5 min at 25 degrees C or for 5, 10, or 15 min at 50 degrees C. The viability loss (in log cycles) following pressurization at 25 degrees C ranged from 0.9 to 3.5 among nine L. monocytogenes strains, 0.7 to 7.8 among seven S. aureus strains, 2.8 to 5.6 among six E. coli O157:H7 strains, and 5.5 to 8.3 among six Salmonella strains. The results show that at 25 degrees C some strains of each species are more resistant to pressure than the others. However, when one resistant and one sensitive strain from each species were pressurized at 345 MPa and 50 degrees C, the population of all except the resistant S. aureus strain was reduced by more than 8 log cycles within 5 min. Viability loss of the resistant S. aureus strain was 6.3 log cycles even after 15 min of pressurization. This shows that strains of food-borne pathogens differ in resistance to hydrostatic pressure (345 MPa) at 25 degrees C, but this difference is greatly reduced at 50 degrees C. Pressurization at 50 degrees C, in place of 25 degrees C, will ensure greater safety of foods.


Site-specific protein double labeling by expressed protein ligation: applications to repeat proteins.
In the last few years, the use of labeled proteins has significantly expanded in the life sciences. Now, labeled proteins are indispensable tools for a wide spectrum of biophysical and chemical biology applications. In particular, the quest for more sophisticated experimental setups requires the development of new synthetic methodology, especially for multiple site-specific labeling. In this paper, we describe a synthetic strategy based on expressed protein ligation to prepare proteins in high purity and homogeneity, in which two different molecular probes are incorporated specifically at any desired position. Proteins are sequentially labeled in solution, with the advantage that a large excess of probes is not required and the labeled fragments are not restricted to peptide synthesis length limitations. This strategy was applied to selectively label a repeat protein with a fluorophores pair in different positions along the protein sequence. The doubly labeled proteins were prepared at high purity and homogeneity, as required for single molecule FRET studies. Remarkably, this approach can be adapted to the introduction of more than two molecular probes.


A Decapeptide Hydrated by Two Waters: Conformers Determined by Theory and Validated by Cold Ion Spectroscopy.
The intrinsic structures of biomolecules in the gas phase may not reflect their native solution geometries. Microsolvation of the molecules bridges the two environments, enabling a tracking of molecular structural changes upon hydration at the atomistic level. We employ density functional calculations to compute a large pool of structures and vibrational spectra for a gas-phase complex, in which a doubly protonated decapeptide, gramicidin S, is solvated by two water molecules. Though most vibrations of this large complex are treated in a harmonic approximation, the water molecules and the vibrations of the host ion coupled to them are locally described by a quantum mechanical vibrational self-consistent field theory with second-order perturbation correction (VSCF-PT2). Guided and validated by the available cold ion spectroscopy data, the computational analysis identifies structures of the three experimentally observed conformers of the complex. They, mainly, differ by the hydration sites, of which the one at the Orn side chain is the most important for reshaping the peptide toward its native structure. The study demonstrates the ability of a quantum chemistry approach that intelligently combines the semiempirical and ab initio computations to disentangle a complex interplay of intra- and intermolecular hydrogen bonds in large molecular systems.


Socioeconomic and environment determinants as predictors of severe malaria in children under 5 years of age admitted in two hospitals in Koudougou district, Burkina Faso: a cross sectional study.
Burkina Faso has a high incidence and death rate of severe malaria, especially for children under 5 years of age. Although the malaria elimination program is a high-priority public health project, finding an effective strategy for managing the problem is a major challenge. Understanding the various factors that contribute to the severity of malaria is essential in designing an effective strategy. In this study, parental and environmental factors associated with severe malaria in Burkinabè children were investigated in two hospitals in Koudougou Health District, Burkina Faso. Between July and September 2012, a cross-sectional study was used to test 510 children under 5 years of age (mean age: 23.5 months) admitted with suspected malaria. Each child was screened using a blood smear to identify whether he or she had severe malaria based on the criteria established by the World Health Organization (WHO). When a child was diagnosed with malaria, either severe or not severe, the parents were interviewed by a trained interviewer using a structured questionnaire. A logistic regression was used to identify the determinants of severe malaria and associated deaths. Of the 510 children having malaria, 201 (39.4%) had severe malaria. Most of the patients (54.9%) lived in rural areas. The main factors associated with severe malaria were low education level of the father, low socioeconomic status [odds ratio (OR)=4.11, 95% confidence interval (CI)=1.44-11.75], delayed treatment [OR=4.53, 95% CI=1.76-11.65], treating children at home as a typical practice when the child has a fever [OR=3.24, 95% CI=1.40-7.51], living in rural area [OR=6.66, 95% CI=3.36-13.22], and living beside a water gathering pond (OR=1.67, 95% CI=1.02-2.74]. Parental and environmental context associated with severe malaria for children under 5 years of age remains a serious public health problem that affects malaria outcomes in resource-limited areas. Promotion of early care is urgently required. Parents should be given information on the risks of not consulting a health facility when children exhibit symptoms of malaria.


Accramycin A, a New Aromatic Polyketide, from the Soil Bacterium, Streptomyces sp. MA37.
Drug-like molecules are known to contain many different building blocks with great potential as pharmacophores for drug discovery. The continued search for unique scaffolds in our laboratory led to the isolation of a novel Ghanaian soil bacterium, Streptomyces sp. MA37. This strain produces many bioactive molecules, most of which belong to carbazoles, pyrrolizidines, and fluorinated metabolites. Further probing of the metabolites of MA37 has led to the discovery of a new naphthacene-type aromatic natural product, which we have named accramycin A 1. This molecule was isolated using an HPLC-photodiode array (PDA) guided isolation process and MS/MS molecular networking. The structure of 1 was characterized by detailed analysis of LC-MS, UV, 1D, and 2D NMR data. Preliminary studies on the antibacterial properties of 1 using Group B Streptococcus (GBS) produced a minimum inhibitory concentration (MIC) of 27 µg/mL. This represents the first report of such bioactivity amongst the naphthacene-type aromatic polyketides, and also suggests the possibility for the further development of potent molecules against GBS based on the accramycin scaffold. A putative acc biosynthetic pathway for accramycin, featuring a tridecaketide-specific type II polyketide synthase, was proposed.


Domain-General and Domain-Specific Patterns of Activity Supporting Metacognition in Human Prefrontal Cortex.
Metacognition is the capacity to evaluate the success of one's own cognitive processes in various domains; for example, memory and perception. It remains controversial whether metacognition relies on a domain-general resource that is applied to different tasks or if self-evaluative processes are domain specific. Here, we investigated this issue directly by examining the neural substrates engaged when metacognitive judgments were made by human participants of both sexes during perceptual and memory tasks matched for stimulus and performance characteristics. By comparing patterns of fMRI activity while subjects evaluated their performance, we revealed both domain-specific and domain-general metacognitive representations. Multivoxel activity patterns in anterior prefrontal cortex predicted levels of confidence in a domain-specific fashion, whereas domain-general signals predicting confidence and accuracy were found in a widespread network in the frontal and posterior midline. The demonstration of domain-specific metacognitive representations suggests the presence of a content-rich mechanism available to introspection and cognitive control.SIGNIFICANCE STATEMENT We used human neuroimaging to investigate processes supporting memory and perceptual metacognition. It remains controversial whether metacognition relies on a global resource that is applied to different tasks or if self-evaluative processes are specific to particular tasks. Using multivariate decoding methods, we provide evidence that perceptual- and memory-specific metacognitive representations coexist with generic confidence signals. Our findings reconcile previously conflicting results on the domain specificity/generality of metacognition and lay the groundwork for a mechanistic understanding of metacognitive judgments.


No permanent reduction in bone mineral density during treatment of polymyalgia rheumatica and temporal arteritis using low dose corticosteroids.
The objective of the study was to examine bone mineral density (BMD) in patients with polymyalgia rheumatica (PMR) or temporal arteritis (TA) currently or previously treated with prednisolone. BMD (using single or dual x-ray absorptiometry) was measured in radius, spine, and hip in 26 currently and 28 previously prednisolone treated patients with PMR (n = 38) or TA (n = 16). The prednisolone treated patients were compared to 30 newly diagnosed PMR (n = 26) or TA patients (n=4) examined prior to start of prednisolone, and 70 healthy controls. No statistically significant differences were found between the groups regarding age, height, weight, and gender. For current users of prednisolone, the mean daily dose was 6.5 mg, the mean cumulative dose 7.7 grams, and for previous users 5.6 mg and 6.6 grams, respectively. No statistically significant differences in BMD at the different measurement sites were found between prednisolone treated patients and the two control groups. Similarly, no significant differences in BMD were found between current and previous users of prednisolone and between the prednisolone treated PMR and TA patients. In conclusion, BMD is not substantially reduced in PMR and TA patients currently or previously treated with mean low dose prednisolone. However, a tendency to a lower BMD was found in PMR/TA patients currently treated with prednisolone and in the prednisolone treated TA patients.


Restricted immunoglobulin variable region (Ig V) gene expression accompanies secondary rearrangements of light chain Ig V genes in mouse plasmacytomas.
The many binding studies of monoclonal immunoglobulin (Ig) produced by plasmacytomas have found no universally common binding properties, but instead, groups of plasmacytomas with specific antigen-binding activities to haptens such as phosphorylcholine, dextrans, fructofuranans, or dinitrophenyl. Subsequently, it was found that plasmacytomas with similar binding chain specificities not only expressed the same idiotype, but rearranged the same light (V(L)) and heavy (V(H)) variable region genes to express a characteristic monoclonal antibody. In this study, we have examined by enzyme-linked immunosorbent assay five antibodies secreted by silicone-induced mouse plasmacytomas using a broader panel of antigens including actin, myosin, tubulin, single-stranded DNA, and double-stranded DNA. We have determined the Ig heavy and light chain V gene usage in these same plasmacytomas at the DNA and RNA level. Our studies reveal: (a) antibodies secreted by plasmacytomas bind to different antigens in a manner similar to that observed for natural autoantibodies; (b) the expressed Ig heavy genes are restricted in V gene usage to the V(H)-J558 family; and (c) secondary rearrangements occur at the light chain level with at least three plasmacytomas expressing both kappa and lambda light chain genes. These results suggest that plasmacytomas use a restricted population of B cells that may still be undergoing rearrangement, thereby bypassing the allelic exclusion normally associated with expression of antibody genes.


The worm that lived: Evolution of rapid aging under high extrinsic mortality revisited.
Organisms age because of the "selection shadow"-the decline of the force of natural selection with age. Seemingly straightforward corollary of this theory is the Medawar-Williams prediction, which maintains that increased extrinsic (non-aging) mortality will result in the evolution of accelerated aging and decreased longevity. Despite its centrality to modern thinking about the ultimate causes of aging, this prediction ignores the fact that mortality is often a non-random process depending on individual condition. Increased condition-dependent mortality inescapably results in increased selection for resistance against the agent of mortality. Provided that resistance to various stressors is commonly associated with increased longevity, the evolutionary outcome is no longer certain. We recently documented this experimentally by showing that populations of Caenorhabditis remanei evolved to live shorter under high extrinsic mortality, but only when mortality was applied haphazardly. On the contrary, when extrinsic mortality was caused by heat-shock, populations experiencing the same rate of increased mortality evolved greater longevities, notwithstanding increased "selection shadow." Intriguingly, stress-resistant and long-lived worms were also more fecund. We discuss these results in the light of recent theoretical developments, such as condition-environment interactions and hyperfunction theory of aging.


Leaf surfaces of Gomphrena spp. (Amaranthaceae) from Cerrado biome.
The leaf structure and micromorphology characterize plant species and reflex its interactions with the environment. Leaf epidermis sculptures aid high transpiration plants on light reflection. The form and distribution of epicuticular wax crystalloids are important to characterize the surface. Aiming to know the micromorphology and the ultrastructure of G. arborescens, G. pohlii and G. virgata, leaves of these Cerrado native species were collected in Brasília, Distrito Federal, Brazil, at the Olympic Center of the Universidade de Brasília and at Reserva Ecológica do Roncador. Leaves of G. globosa, an Indian native species, were also studied for comparison. Leaves were fractionated, fixed and treated for observation under optical and scanning electron microscope. A description of the leaf epidermis is provided, along with some quantitative data to help the species taxonomy and support future studies on their physiology: all species are amphistomatic and have Stomatal Index between 7.27 and 18.99. The Gomphrena spp. studied have epicuticular wax platelets and wax sculptures over their larger trichome, which are relevant for their taxonomy. Over the Cerrado species cuticle, epicuticular wax is damaged by fungi hyphae development. The presence of epicuticular wax on Gomphrena spp. leaves corroborates the phylogenetical alliance between Amaranthaceae and Chenopodiaceae.


Equivalence of lauric acid and glycerol monolaurate as inhibitors of signal transduction in Staphylococcus aureus.
Glycerol monolaurate (GML) inhibits the expression of virulence factors in Staphylococus aureus and the induction of vancomycin resistance in Enterococcus faecalis, presumably by blocking signal transduction. Although GML is rapidly hydrolyzed by bacteria, one of the products, lauric acid, has identical inhibitory activity and is metabolized much more slowly. At least four distinct GML-hydrolyzing activities are identified in S. aureus: the secreted Geh lipase, residual supernatant activity in a geh-null mutant strain, a novel membrane-bound esterase, and a cytoplasmic activity.


Cigarette smoking and its orthopedic consequences.
Cigarette smoking and its ramifications are coming under increasing scrutiny in the field of orthopedic surgery. Smoking has been implicated in impeding bone metabolism and fracture repair, and increasing the rate of postoperative infection and the incidence of nonunion. This article reviews the current body of knowledge on these topics, as well as the potential adverse effects of smoking on wound healing and microsurgical procedures. An in-depth discussion on the pathophysiologic mechanisms of nicotine is also included.


Arabidopsis SWC4 Binds DNA and Recruits the SWR1 Complex to Modulate Histone H2A.Z Deposition at Key Regulatory Genes.
Deposition of the H2A.Z histone variant by the SWR1 complex (SWR1-C) in regulatory regions of specific loci modulates transcription. Characterization of mutations in Arabidopsis thaliana homologs of yeast SWR1-C has revealed a role for H2A.Z exchange in a variety of developmental processes. Nevertheless, the exact composition of plant SWR1-C and how it is recruited to target genes remains to be established. Here we show that SWC4, the Arabidopsis homolog of yeast SANT domain protein Swc4/Eaf2, is a DNA-binding protein that interacts with SWR1-C subunits. We demonstrate that the swc4-1 knockout mutant is embryo-lethal, while SWC4 RNAi knockdown lines display pleiotropic phenotypic alterations in vegetative and reproductive traits, including acceleration of flowering time, indicating that SWC4 controls post-embryonic processes. Transcriptomic analyses and genome-wide profiling of H2A.Z indicate that SWC4 represses transcription of a number of genes, including the floral integrator FT and key transcription factors, mainly by modulating H2A.Z deposition. Interestingly, SWC4 silencing does not affect H2A.Z deposition at the FLC locus nor expression of this gene, a master regulator of flowering previously shown to be controlled by SWR1-C. Importantly, we find that SWC4 recognizes specific AT-rich DNA elements in the chromatin regions of target genes and that SWC4 silencing impairs SWR1-C binding at FT. Collectively, our data suggest that SWC4 regulates plant growth and development by aiding SWR1-C recruitment and modulating H2A.Z deposition.


Analysis of potential changes in seriousness of influenza A and B viruses in Hong Kong from 2001 to 2011.
Continued monitoring of the seriousness of influenza viruses is a public health priority. We applied time-series regression models to data on cardio-respiratory mortality rates in Hong Kong from 2001 to 2011. We used surveillance data on outpatient consultations for influenza-like illness, and laboratory detections of influenza types/subtypes to construct proxy measures of influenza activity. In the model we allowed the regression coefficients for influenza to drift over time, and adjusted for temperature and humidity. The regression coefficient for influenza A(H3N2) increased significantly in 2005. The regression coefficients for influenza A(H1N1) and B were relatively stable over the period. Our model suggested an increase in seriousness of A(H3N2) in 2005, the year after the appearance of the A/Fujian/411/2002(H3N2)-like virus when the drifted A/California/7/2004(H3N2)-like virus appeared. Ongoing monitoring of mortality and influenza activity could permit identification of future changes in seriousness of influenza virus infections.


α-Synucleinopathy associated c-Abl activation causes p53-dependent autophagy impairment.
Studies link c-Abl activation with the accumulation of pathogenic α-synuclein (αS) and neurodegeneration in Parkinson's disease (PD). Currently, c-Abl, a tyrosine kinase activated by cellular stress, is thought to promote αS pathology by either directly phosphorylating αS or by causing autophagy deficits. αS overexpressing transgenic (Tg) mice were used in this study. A53T Tg mice that express high levels of human mutant A53TαS under the control of prion protein promoter. Two different approaches were used in this study. Natural aging and seeding model of synucleinopathy. In seeding model, intracortical/intrastriatal (IC/IS) stereotaxic injection of toxic lysates was done using tissue lysates from end-stage symptomatic mice. In this study, nilotinib and pifithrin-α was used as a c-Abl and p53 inhibitor, respectively. Both Tg and non-transgenic (nTg) mice from each group were subjected to nilotinib (10 mg/kg) or vehicle (DMSO) treatment. Frozen brain tissues from PD and control human cases were analyzed. In vitro cells study was implied for c-Abl/p53 genetic manipulation to uncover signal transduction. Herein, we show that the pathologic effects of c-Abl in PD also involve activation of p53, as c-Abl activation in a transgenic mouse model of α-synucleinopathy (TgA53T) and human PD cases are associated with the increased p53 activation. Significantly, active p53 in TgA53T neurons accumulates in the cytosol, which may lead to inhibition of autophagy. Thus, we hypothesized that c-Abl-dependent p53 activation contributes to autophagy impairment in α-synucleinopathy. In support of the hypothesis, we show that c-Abl activation is sufficient to inhibit autophagy in p53-dependent manner. Moreover, inhibition of either c-Abl, using nilotinib, or p53, using pifithrin-α, was sufficient to increase autophagic flux in neuronal cells by inducing phosphorylation of AMP-activated kinase (AMPK), ULK1 activation, and down-regulation of mTORC1 signaling. Finally, we show that pharmacological attenuation of c-Abl activity by nilotinib treatment in the TgA53T mouse model reduces activation of p53, stimulates autophagy, decreases accumulation αS pathology, and delays disease onset. Collectively, our data show that c-Abl activation by α-synucleinopathy causes p53 dependent autophagy deficits and both c-Abl and p53 represent therapeutic target for PD.


Chemokine and chemokine receptor expression in a novel human mesangial cell line.
Chemokines are thought to play a pivotal role in mediating the selective migration of leukocytes into sites of tissue injury. The local production of chemokines by mesangial cells (MC) has been linked to inflammatory processes within the glomerulus. To study the chemokine biology of human MC, an immortalized human MC line was generated and then chemokine and chemokine receptor expression was examined in response to various proinflammatory stimuli. The results show that human MC have a specific and limited repertoire of chemokine expression. The stimulus-specific regulation of the chemokines monocyte chemoattractant protein- (MCP- 1), regulated upon activation, normal T cell expressed and secreted (RANTES), interleukin-8 (IL-8), and IP-10 was demonstrated using RNase protection assays. Transcripts for the chemokines MIP-1alpha, MIP-1beta, I-309, or lymphotactin could not be detected. The expression of CC chemokine receptors was investigated by reverse transcription-PCR and RNase protection assays. MC stimulated with interferon-gamma (IFN-gamma) expressed mRNA for the chemokine receptor CCR1. The expression could be further increased by activating the cells with a combination of tumor necrosis factor-a (TNF-alpha), IL-1beta, and IFN-gamma. Under these conditions, no mRNA for CCR2, CCR3, CCR4, CCR5, or CCR8 was detected. A comparison of the immortalized human mesangial cells with primary cells showed identical expression patterns of chemokine receptors. To demonstrate functional activity of chemokine receptors expressed by human MC, chemotaxis assays were performed. MC stimulated with a combination of TNF-alpha, IL-1beta, and IFN-gamma, but not unstimulated MC, migrated toward a RANTES gradient. Eotaxin did not enhance the migratory activity of human MC. In summary, a novel human mesangial cell line was established and the pattern of chemokine expression was examined. For the first time, the inducible expression of functionally active CCR1 by human MC was shown.


Does the Configuration at the Metal Matter in Noyori-Ikariya Type Asymmetric Transfer Hydrogenation Catalysts?
Noyori-Ikariya type [(arene)RuCl(TsDPEN)] (TsDPEN, sulfonated diphenyl ethylenediamine) complexes are widely used C=O and C=N reduction catalysts that produce chiral alcohols and amines via a key ruthenium-hydride intermediate that determines the stereochemistry of the product. Whereas many details about the interactions of the pro-chiral substrate with the hydride complex and the nature of the hydrogen transfer from the latter to the former have been investigated over the past 25 years, the role of the stereochemical configuration at the stereogenic ruthenium center in the catalysis has not been elucidated so far. Using operando FlowNMR spectroscopy and nuclear Overhauser effect spectroscopy, we show the existence of two diastereomeric hydride complexes under reaction conditions, assign their absolute configurations in solution, and monitor their interconversion during transfer hydrogenation catalysis. Configurational analysis and multifunctional density functional theory (DFT) calculations show the λ-(R,R)S Ru configured [(mesitylene)RuH(TsDPEN)] complex to be both thermodynamically and kinetically favored over its λ-(R,R)R Ru isomer with the opposite configuration at the metal. Computational analysis of both diastereomeric catalytic manifolds show the major λ-(R,R)S Ru configured [(mesitylene)RuH(TsDPEN)] complex to dominate asymmetric ketone reduction catalysis with the minor λ-(R,R)R Ru [(mesitylene)RuH(TsDPEN)] stereoisomer being both less active and less enantioselective. These findings also hold true for a tethered catalyst derivative with a propyl linker between the arene and TsDPEN ligands and thus show enantioselective transfer hydrogenation catalysis with Noyori-Ikariya complexes to proceed via a lock-and-key mechanism.


Comparison of two doses of hypobaric bupivacaine in unilateral spinal anesthesia for hip fracture surgery: 5 mg versus 7.5 mg.
Hip fracture is a frequent and severe disease. Its prognosis depends on the perioperative hemodynamic stability which can be preserved by the unilateral spinal anesthesia especially with low doses of local anesthetics. This study aims to compare the efficacy and hemodynamic stability of two doses of hypobaric bupivacaine (7.5 mg vs 5 mg) in unilateral spinal anesthesia. In this prospective, randomized, double-blind study, 108 patients scheduled for hip fracture surgery under unilateral spinal anesthesia were enrolled to receive either 5 mg (group 1) or 7.5 mg (group 2) of hypobaric bupivacaine. Spinal anesthesia was performed in lateral position. Patients' socio-demographic characteristics, hemodynamic profile, sensory and motor blocks parameters were recorded. Both groups were comparable regarding to demographic data. Two cases of failure occurred in group 1 and one case in group 2 corresponding to a comparable efficiency rates (96.29% and 98.14% respectively; p = 0.5). A higher mean onset and lower mean regression times of sensory block were significantly noted in group 1 (7.79±3.76 min vs 5.75±2.35 min, p < 0.001 and 91.29±31.55 min vs 112.77±18.77 min, p <0.001 respectively). Incidence of bilateralization (29.62% vs 87.03%, p < 0.001), incidence of hypotensive episodes (59.25% vs 92.59%, p < 0.001) and vascular loading (1481.48±411.65 ml vs 2111.11±596.10 ml, p < 0.001) were significantly higher in group 2. The dosage of 5mg of hypobaric bupivacaine in unilateral spinal anesthesia is as effective as the dosage of 7.5 mg with lower bilateralization incidence and better hemodynamic stability.


Impact of computational approaches in the fight against COVID-19: an AI guided review of 17 000 studies.
SARS-CoV-2 caused the first severe pandemic of the digital era. Computational approaches have been ubiquitously used in an attempt to timely and effectively cope with the resulting global health crisis. In order to extensively assess such contribution, we collected, categorized and prioritized over 17 000 COVID-19-related research articles including both peer-reviewed and preprint publications that make a relevant use of computational approaches. Using machine learning methods, we identified six broad application areas i.e. Molecular Pharmacology and Biomarkers, Molecular Virology, Epidemiology, Healthcare, Clinical Medicine and Clinical Imaging. We then used our prioritization model as a guidance through an extensive, systematic review of the most relevant studies. We believe that the remarkable contribution provided by computational applications during the ongoing pandemic motivates additional efforts toward their further development and adoption, with the aim of enhancing preparedness and critical response for current and future emergencies.


Environmental exposure assessment in European birth cohorts: results from the ENRIECO project.
Environmental exposures during pregnancy and early life may have adverse health effects. Single birth cohort studies often lack statistical power to tease out such effects reliably. To improve the use of existing data and to facilitate collaboration among these studies, an inventory of the environmental exposure and health data in these studies was made as part of the ENRIECO (Environmental Health Risks in European Birth Cohorts) project. The focus with regard to exposure was on outdoor air pollution, water contamination, allergens and biological organisms, metals, pesticides, smoking and second hand tobacco smoke (SHS), persistent organic pollutants (POPs), noise, radiation, and occupational exposures. The review lists methods and data on environmental exposures in 37 European birth cohort studies. Most data is currently available for smoking and SHS (N=37 cohorts), occupational exposures (N=33), outdoor air pollution, and allergens and microbial agents (N=27). Exposure modeling is increasingly used for long-term air pollution exposure assessment; biomonitoring is used for assessment of exposure to metals, POPs and other chemicals; and environmental monitoring for house dust mite exposure assessment. Collaborative analyses with data from several birth cohorts have already been performed successfully for outdoor air pollution, water contamination, allergens, biological contaminants, molds, POPs and SHS. Key success factors for collaborative analyses are common definitions of main exposure and health variables. Our review emphasizes that such common definitions need ideally be arrived at in the study design phase. However, careful comparison of methods used in existing studies also offers excellent opportunities for collaborative analyses. Investigators can use this review to evaluate the potential for future collaborative analyses with respect to data availability and methods used in the different cohorts and to identify potential partners for a specific research question.


Hybrid therapy of atrial fibrillation: algorithms and outcome.
Rhythm control methods have not shown superior outcomes to rate control strategies in atrial fibrillation. Newer approaches to rhythm control employ "hybrid" therapies combining pharmacologic and non pharmacologic interventions. Pathophysiologic insights into mechanisms of atrial fibrillation (AF) suggest that arrhythmogenesis is due to interactions of multiple triggering rhythms and a complex electrophysiologic substrate resulting in the emergence of multiple tachyarrhythmias, often in disparate locations that may coexist in time. Thus, an "hybrid" therapy prescription is more likely to address several of the etiologic factors culminating in clinical AF. Results of pilot clinical studies of hybrid therapy are encouraging and involve drugs, devices and ablation techniques in varying permutations. Hybrid therapy algorithms using right heart procedures can improve efficacy with potentially lower risk. Considerations in implementation of these algorithms include staged or simultaneous interventions and a right versus left heart strategy. The parallel with the current coronary disease management paradigm is obvious and relevant.


Case Report of Focal Epithelial Hyperplasia (Heck's Disease) with Polymerase Chain Reaction Detection of Human Papillomavirus 13.
Focal epithelial hyperplasia (FEH), or Heck's disease, is an uncommon benign proliferation of oral mucosa caused by the human papillomavirus (HPV), particularly subtypes 13 and 32. The disease typically presents in young Native American patients and is characterized by multiple asymptomatic papules and nodules on the oral mucosa, lips, tongue, and gingiva. The factors that determine susceptibility to FEH are unknown, but the ethnic and geographic distribution of FEH suggests that genetic predisposition, particularly having the human lymphocytic antigen DR4 type, may be involved in pathogenesis. We report a case of FEH with polymerase chain reaction detection of HPV13 in a healthy 11-year-old Hispanic girl and discuss the current understanding of disease pathogenesis, susceptibility, and treatment.


Does serum butyrylcholinesterase level determine the severity and mortality of COVID-19 pneumonia?: Prospective study.
The WHO emphasized the importance of knowing the risk factors for the severity of the disease in the COVID-19 pandemic. Our aim in this study was to determine the relationship between serum Butyrylcholinesterase (BChE) level, which is rapidly affected by inflammation, and the severity of COVID-19 pneumonia and mortality. Patients diagnosed with COVID-19 pneumonia between March and May 2021 were included in the study. The patients were divided into two groups as severe and mild to moderate pneumonia according to the WHO's guidelines. Serum BChE levels were studied by ELISA method from the blood samples taken from the patients on the day of hospitalization. The severity of the disease and other factors affecting hospital mortality were also evaluated. 147 patients with COVID-19 pneumonia were included in this study. Of these patients, 58% had severe pneumonia and 42% had mild to moderate pneumonia. The BChE level was median 13 (IQR: 11.2-21.5)ng/ml in patients with severe COVID-19 pneumonia and median 20 (IQR: 10-35.7)ng/ml in patients with mild to moderate pneumonia (p: 0.001). Hospital with mortality rate was higher in patients with low BChE levels. However, statistically, BChE hasn't associated mortality in COVID-19 pneumonia [OR 1.002 (0.957-1.049) p: 0.490]. CRP, procalcitonin, lactate, and D-dimer levels were associated mortality in COVID-19 pneumonia. Being not statistically significant, the mortality rate was higher in patients with low BChE levels. BChE level is an important marker in determining the severity of COVID-19 pneumonia. Early prediction of the severity of COVID-19 pneumonia will enable early planning of the treatment process.


Genome ploidy in different stages of the Giardia lamblia life cycle.
The early diverging eukaryotic parasite Giardia lamblia is unusual in that it contains two apparently identical nuclei in the vegetative trophozoite stage. We have determined the nuclear and cellular genome ploidy of G. lamblia cells during all stages of the life cycle. During vegetative growth, the nuclei cycle between a diploid (2N) and tetraploid (4N) genome content and the cell, consequently, cycles between 4N and 8N. Stationary phase trophozoites arrest in the G2 phase with a ploidy of 8N (two nuclei, each with a 4N ploidy). On its way to cyst formation, a G1 trophozoite goes through two successive rounds of chromosome replication without an intervening cell division event. Fully differentiated cysts contain four nuclei, each with a ploidy of 4N, resulting in a cyst ploidy of 16N. The newly excysted cell, for which we suggest the term 'excyzoite', contains four nuclei (cellular ploidy 16N). In a reversal of the events occurring during encystation, the excyzoite divides twice to form four trophozoites containing two diploid nuclei each. The formation of multiple cells from a single cyst is likely to be one of the main reasons for the low infectious doses of G. lamblia.


Artificial neural networks and center-of-pressure modeling: a practical method for sensorimotor-degradation assessment.
Numerous methods for studying the prevention of falls and age-related sensorimotor degradation have been proposed and tested. Some approaches are too impractical to use with seniors or too expensive for practitioners. Practitioners desire a simple, reliable technique. The goals of this research were to develop such an approach and to apply it in exploring the effect of Tai Chi on age-related sensorimotor degradation. The method employed artificial-neural-network (ANN) models trained by using individuals' center-of-pressure (COP) measurements and age. Ninety-six White and Chinese adults without Tai Chi training were tested. In contrast, a third group, Chinese seniors with Tai Chi training, was tested to ascertain any influence from Tai Chi on sensorimotor aging. This study supported ANN technology with COP data as a feasible tool in the exploration of sensorimotor degradation and demonstrated that Tai Chi slowed down the effects of sensorimotor aging.


A three-dimensional edge operator.
Modern scanning techniques, such as computed tomography, have begun to produce true three-dimensional imagery of internal structures. The first stage in finding structure in these images, like that for standard two-dimensional images, is to evaluate a local edge operator over the image. If an edge segment in two dimensions is modeled as an oriented unit line segment that separates unit squares (i.e., pixels) of different intensities, then a three-dimensional edge segment is an oriented unit plane that separates unit volumes (i.e., voxels) of different intensities. In this correspondence we derive an operator that finds the best oriented plane at each point in the image. This operator, which is based directly on the 3-D problem, complements other approaches that are either interactive or heuristic extensions of 2-D techniques.


Replication stress affects the fidelity of nucleosome-mediated epigenetic inheritance.
The fidelity of epigenetic inheritance or, the precision by which epigenetic information is passed along, is an essential parameter for measuring the effectiveness of the process. How the precision of the process is achieved or modulated, however, remains largely elusive. We have performed quantitative measurement of epigenetic fidelity, using position effect variegation (PEV) in Schizosaccharomyces pombe as readout, to explore whether replication perturbation affects nucleosome-mediated epigenetic inheritance. We show that replication stresses, due to either hydroxyurea treatment or various forms of genetic lesions of the replication machinery, reduce the inheritance accuracy of CENP-A/Cnp1 nucleosome positioning within centromere. Mechanistically, we demonstrate that excessive formation of single-stranded DNA, a common molecular abnormality under these conditions, might have correlation with the reduction in fidelity of centromeric chromatin duplication. Furthermore, we show that replication stress broadly changes chromatin structure at various loci in the genome, such as telomere heterochromatin expanding and mating type locus heterochromatin spreading out of the boundaries. Interestingly, the levels of inheritable expanding at sub-telomeric heterochromatin regions are highly variable among independent cell populations. Finally, we show that HU treatment of the multi-cellular organisms C. elegans and D. melanogaster affects epigenetically programmed development and PEV, illustrating the evolutionary conservation of the phenomenon. Replication stress, in addition to its demonstrated role in genetic instability, promotes variable epigenetic instability throughout the epigenome.


Epidemiology of Hip Flexor and Hip Adductor Strains in National Collegiate Athletic Association Athletes, 2009/2010-2014/2015.
Little research has examined the rates and patterns of hip flexor or hip adductor strains in student-athletes in the National Collegiate Athletic Association (NCAA). To describe the epidemiology of hip flexor and adductor strains in NCAA athletes during the 2009/2010-2014/2015 academic years. Descriptive epidemiology study. Rates and patterns of hip flexor and adductor strains in collegiate sports were examined in a convenience sample of NCAA varsity teams from 25 sports. Rates and distributions of strains by mechanism, recurrence, and participation restriction time were examined. Injury rate ratios (IRRs) and proportion ratios were calculated to compare rates within and between sports by event type, sex, mechanism, recurrence, and participation restriction time. A total of 770 hip flexor and 621 hip adductor strains were reported, resulting in overall injury rates of 1.60 and 1.29 per 10,000 athlete-exposures (AEs), respectively. In men, the rate of hip flexor strains was 1.81 per 10,000 AEs, and that for hip adductor strains was 1.71 per 10,000 AEs. In women, the rate of hip flexor strains was 1.59 per 10,000 AEs, and the rate of hip adductor strains was 1.15 per 10,000 AEs. The highest rates of strains were found in men's soccer and men's ice hockey (range, 2.47-3.77 per 10,000 AEs). Most hip flexor and hip adductor strains occurred in practice, but both had higher rates in competition. In sex-comparable sports, hip flexor strain rates did not differ between the sexes (IRR = 1.14; 95% CI, 0.96-1.36), but the rate of hip adductor strains was higher in men than women (IRR = 1.49; 95% CI, 1.22-1.81). Noncontact was the most common mechanism for both types of strains (hip flexor strains, 59.4%; hip adductor strains, 62.5%); 10.1% of hip flexor strains and 11.1% of hip adductor strains were recurrent. The highest rates of recurrence of both types of strain were found in men's and women's ice hockey (range, 16.0%-30.6%). Most hip flexor and hip adductor strains resulted in <1 week of participation restriction (hip flexor strains, 83.8%; hip adductor strains, 82.9%). The NCAA sports with the highest rates of hip flexor and hip adductor strains were men's soccer and men's ice hockey. In sex-comparable sports, men had a higher rate of hip adductor, but not hip flexor, strains. Recurrence rates were remarkably high in ice hockey. Male sports teams, especially soccer and ice hockey, should place an emphasis on prevention programs for hip adductor strains. Secondary prevention programs involving thorough rehabilitation and strict return-to-play criteria should be developed and implemented to curb the high recurrence rate of these injuries, particularly in ice hockey.


Potential fate of ingested Lactobacillus plantarum and its occurrence in human feces.
Lactobacillus plantarum has been used in human clinical trials to promote beneficial effects in the immune system, to alleviate intestinal disorders, and to reduce the risk of cardiovascular disease. It is also involved in many fermentation processes in the food industry. However, information on the fate of ingested L. plantarum is limited. In this study, 61 subjects received daily doses of fermented milk containing 2 × 10(11) cells of L. plantarum Lp115 for different periods of time. The target microorganism was monitored in the fecal microbiota via quantitative PCR (qPCR). L. plantarum was detected and quantified in all of the subjects during the ingestion periods. The differences between the L. plantarum levels at time zero and during all the different ingestion periods were statistically significant (P = 0.001). However, at 15 and 45 days after discontinuing supplementation, the number of lactobacilli was reduced to the baseline level (those at time zero). A longer period with L. plantarum in the diet did not result in increased levels of this bacterium in the stool, based on postconsumption evaluations (P = 0.001). The qPCR method was specific and sensitive for L. plantarum quantification in such a complex microbial environment as the gastrointestinal tract.


[Pathogenetic justification for the surgeon's active procedure in acute gastrointestinal hemorrhages of peptic origin].
The relationship between the disorders in the indices of the internal medium of the organism and the volume of blood loss, the number of recurrent hemorrhages, the length of the posthemorrhagic period was established on the basis of experimental, laboratory, instrumental investigations and the experience of treatment of 1987 patients with acute gastro-intestinal hemorrhages. The advantages of active methods and the early urgent operation over other kinds of treatment were supported by the immediate results of operations in 415 patients with profuse gastro-intestinal hemorrhage of ulcerous etiology. The terms and results of operations were shown to be in direct dependence. The mortality rate after the operations performed within 24 hours was 1%.


Understanding the Burden of 30-Day Readmission in Patients With Both Primary and Secondary Diagnoses of Heart Failure: Causes, Timing, and Impact of Co-Morbidities.
Although efforts to reduce 30-day readmission rates have mainly focused on patients with heart failure (HF) as a primary diagnosis at index hospitalization, patients with HF as a secondary diagnosis remain common, costly, and understudied. This study aimed to determine the incidence, etiology, and patterns of 30-day readmissions after discharge for HF as a primary and secondary diagnosis and investigate the impact of co-morbidities on HF readmission. The National Readmission Database from 2014 to 2016 was used to identify HF patients with a linked 30-day readmission. Patient and hospital characteristics, admission features, and Elixhauser-related co-morbidities were compared between the 2 groups. Readmitted patients in both groups were younger, male, with lower household income, higher mortality risk, and higher hospitalization costs. Over 60% of readmissions were for reasons other than HF, and greater than 1/3 had more than 2 readmissions within 30 days, with a median time to readmission of 12 days. Both cohorts had high readmission rates and high rates of readmission for causes other than HF. Our findings suggest that efforts to reduce 30-day readmission rates should be extended to patients with secondary HF diagnosis, with surveillance extending to 2 weeks postdischarge to identify patients at risk.


Effect of aminoguanidine on ischemia-reperfusion induced myocardial injury in rats.
Myocardial ischemia-reperfusion (MI/R) has been implicated in the induction of inducible nitric oxide synthase (iNOS) that leads to increase production of nitric oxide (NO). Recently, excessive production of NO has been involved in causing myocardial injury. In our in vivo model, we examined the effects of aminoguanidine (AMG), a known iNOS inhibitor, on percentage infarct size in anaesthetized rats. A total of 14 rats were equally divided into two groups (n = 7 in each group). To produce myocardial necrosis, the left main coronary artery was occluded for 30 min, followed by 120 min of reperfusion, in anesthetized rats. AMG (200 mg kg(-1)) was given intravenously 10 min before occlusion. The volume of infarct size and the risk zone were determined by planimentry of each tracing and multiplying by the slice thickness. Infarct size was normalized by expressing it as a percentage of the area at risk. Hemodynamic parameters were measured via the left carotid artery. Compared to MI/R group, whereas AMG administration elevated mean arterial blood pressure, statistically reduced the myocardial infarct size (21+/- 1 and 14+/- 4%, respectively) and infract size/risk zone (53+/- 3 and 37+/- 5%, respectively) in rat model of ischemia-reperfusion. In conclusion, this study indicates that iNOS inhibitor, AMG, show reduction in NO's side effect in I/R injury.


Incremental value of the preoperative echocardiogram to predict mortality and major morbidity in coronary artery bypass surgery.
Although echocardiography is commonly performed before coronary artery bypass surgery, there has yet to be a study examining the incremental prognostic value of a complete echocardiogram. Patients undergoing isolated coronary artery bypass surgery at 2 hospitals were divided into derivation and validation cohorts. A panel of quantitative echocardiographic parameters was measured. Clinical variables were extracted from the Society of Thoracic Surgeons database. The primary outcome was in-hospital mortality or major morbidity, and the secondary outcome was long-term all-cause mortality. The derivation cohort consisted of 667 patients with a mean age of 67.2±11.1 years and 22.8% females. The following echocardiographic parameters were found to be optimal predictors of mortality or major morbidity: severe diastolic dysfunction, as evidenced by restrictive filling (odds ratio, 2.96; 95% confidence interval, 1.59-5.49), right ventricular dysfunction, as evidenced by fractional area change <35% (odds ratio, 3.03; 95% confidence interval, 1.28-7.20), or myocardial performance index >0.40 (odds ratio, 1.89; 95% confidence interval, 1.13-3.15). These results were confirmed in the validation cohort of 187 patients. When added to the Society of Thoracic Surgeons risk score, the echocardiographic parameters resulted in a net improvement in model discrimination and reclassification with a change in c-statistic from 0.68 to 0.73 and an integrated discrimination improvement of 5.9% (95% confidence interval, 2.8%-8.9%). In the Cox proportional hazards model, right ventricular dysfunction and pulmonary hypertension were independently predictive of mortality over 3.2 years of follow-up. Preoperative echocardiography, in particular right ventricular dysfunction and restrictive left ventricular filling, provides incremental prognostic value in identifying patients at higher risk of mortality or major morbidity after coronary artery bypass surgery.


Improving the protein activity and stability under acidic conditions via site-specific conjugation of a pH-responsive polyelectrolyte.
Maintaining the protein activity and stability under acidic conditions is important in bioengineering and biomedical applications. Polyelectrolyte conjugation as a means of stabilizing proteins has received much recent attention. Retention of protein activity, and especially, improvement of protein stability by minimizing the number of polymer chains in the conjugate, as well as by choosing the optimal site for conjugation, is critical in practical applications. In this research, the cationic polyelectrolyte poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) was conjugated to the inorganic pyrophosphatase (PPase) site specifically. Conjugation of pDMAEMA to the specific site N124 on the protein surface led to a significant increase in activity at acidic pH. At pH 4.0, the activity of the pDMAEMA-conjugated protein was increased 3-fold relative to the unconjugated one. Dynamic light scattering (DLS) measurements showed that the aggregation state of the protein depended on the polymer charge as the pH was varied. Protein aggregation at low pH was prevented by pDMAEMA conjugation, resulting in an increase in protein stability under acidic conditions. The conjugate retained 60% of its initial activity after 4 h at pH 4.0, whereas the unconjugated protein lost 40% of its initial activity within 15 min at this pH. These results suggest an approach for preserving the protein activity and stability at low pH based on site-specific polyelectrolyte conjugation to the protein surface, thereby providing a new strategy for expanding the use of proteins in an acidic environment.


Species-level deconvolution of metagenome assemblies with Hi-C-based contact probability maps.
Microbial communities consist of mixed populations of organisms, including unknown species in unknown abundances. These communities are often studied through metagenomic shotgun sequencing, but standard library construction methods remove long-range contiguity information; thus, shotgun sequencing and de novo assembly of a metagenome typically yield a collection of contigs that cannot readily be grouped by species. Methods for generating chromatin-level contact probability maps, e.g., as generated by the Hi-C method, provide a signal of contiguity that is completely intracellular and contains both intrachromosomal and interchromosomal information. Here, we demonstrate how this signal can be exploited to reconstruct the individual genomes of microbial species present within a mixed sample. We apply this approach to two synthetic metagenome samples, successfully clustering the genome content of fungal, bacterial, and archaeal species with more than 99% agreement with published reference genomes. We also show that the Hi-C signal can secondarily be used to create scaffolded genome assemblies of individual eukaryotic species present within the microbial community, with higher levels of contiguity than some of the species' published reference genomes.


An identifiable molluscan neuron responds to changes in earth-strength magnetic fields.
Diverse animals can orient using geomagnetic cues, but little is known about the neurophysiological mechanisms that underlie magnetic field detection. The marine mollusc Tritonia diomedea (Bergh) has a magnetic sense and its nervous system is amenable to cellular-level electrophysiological analysis. In a semi-intact whole-animal preparation, intracellular recordings from the large, visually identifiable neurons left pedal 5 (LPe5) and right pedal 5 (RPe5) in the brain of Tritonia revealed enhanced electrical activity in response to changes in ambient earth-strength magnetic fields. No such changes in activity were observed in approximately 50 other neurons subjected to identical magnetic stimuli. The responses of LPe5 were characterized by increases in spiking frequency occurring about 6-16 min after the ambient magnetic field had been rotated to a new position. The response was abolished when the brain had been isolated from the periphery of the animal by severing nerves, a procedure that also transected prominent neurites of LPe5. We hypothesize that LPe5 is one component of a neural circuit mediating detection of the earth's magnetic field or orientation to it.


Lipopolysaccharide-Elicited TSLPR Expression Enriches a Functionally Discrete Subset of Human CD14+ CD1c+ Monocytes.
Thymic stromal lymphopoietin (TSLP) is a cytokine produced mainly by epithelial cells in response to inflammatory or microbial stimuli and binds to the TSLP receptor (TSLPR) complex, a heterodimer composed of TSLPR and IL-7 receptor α (CD127). TSLP activates multiple immune cell subsets expressing the TSLPR complex and plays a role in several models of disease. Although human monocytes express TSLPR and CD127 mRNAs in response to the TLR4 agonist LPS, their responsiveness to TSLP is poorly defined. We demonstrate that TSLP enhances human CD14+ monocyte CCL17 production in response to LPS and IL-4. Surprisingly, only a subset of CD14+ CD16- monocytes, TSLPR+ monocytes (TSLPR+ mono), expresses TSLPR complex upon LPS stimulation in an NF-κB- and p38-dependent manner. Phenotypic, functional, and transcriptomic analysis revealed specific features of TSLPR+ mono, including higher CCL17 and IL-10 production and increased expression of genes with important immune functions (i.e., GAS6, ALOX15B, FCGR2B, LAIR1). Strikingly, TSLPR+ mono express higher levels of the dendritic cell marker CD1c. This evidence led us to identify a subset of peripheral blood CD14+ CD1c+ cells that expresses the highest levels of TSLPR upon LPS stimulation. The translational relevance of these findings is highlighted by the higher expression of TSLPR and CD127 mRNAs in monocytes isolated from patients with Gram-negative sepsis compared with healthy control subjects. Our results emphasize a phenotypic and functional heterogeneity in an apparently homogeneous population of human CD14+ CD16- monocytes and prompt further ontogenetic and functional analysis of CD14+ CD1c+ and LPS-activated CD14+ CD1c+ TSLPR+ mono.


Site-directed mutations altering methyl-accepting residues of a sensory transducer protein.
The Trg protein is one of a family of transducer proteins that mediate chemotactic response in Escherichia coli. Transducers are methyl-accepting proteins that gain or lose methyl esters on specific glutamyl residues during sensory adaptation. In this study, the significance of multiple sites of methylation on transducer proteins was addressed by using oligonucleotide-directed, site-specific mutagenesis to substitute an alanyl residue at each of the five methyl-accepting sites in Trg. The resulting collection of five mutations, each inactivating a single site, was analyzed for effects on covalent modification at the remaining sites on Trg and for the ability of the altered proteins to mediate sensory adaptation. Most of the alanyl substitutions had substantial biochemical effects, enhancing or reducing methyl-accepting activity of other sites, including one case of activation of a site not methylated in wild-type protein. Analysis of the altered proteins provided explanations for many features of the complex pattern of electrophoretic forms exhibited by Trg. The mutant proteins were less efficient than normal Trg in mediating adaptation. Correlation of biochemical and behavioral data indicated that reduction in the number of methyl-accepting sites on the transducer lengthened the time required to reach an adapted state.


[Amalgam fillings and T-lymphocyte changes].
Dental amalgam and nickel alloys have been considered quite safe. Previous authors reported the effect of dental amalgam and nickel alloys on human T-lymphocytes modifications after amalgam dental fillings, into dose-dependence of any modifications and into possible temporary. Eight patients were subjected to dental care with amalgam dental fillings. Drawings of blood were executed at start, fifteen days after late fillings and two months later. The results about modifications of T-lymphocytes were not univocal. We believe, at now, that temporary modifications of the immunity seem to be related to a cytotoxic mechanism.


Pseudomonas bacteremia. Retrospective analysis of 410 episodes.
We reviewed 410 episodes of Pseudomonas bacteremia occurring in patients with cancer during a ten-year period. Pseudomonas bacteremia was most common among patients with acute leukemia. The majority of patients acquired their infections in the hospital, and 51% had received antibiotic therapy for other presumed or proved infection during the preceding week. Shock occurred in 33%, and 32% had concomitant pneumonia. The overall cure rate was 62%; it was 67% for patients receiving appropriate antibiotics but only 14% for those receiving inappropriate antibiotics. A one- to two-day delay in the administration of appropriate antibiotic therapy reduced the cure rate from 74% to 46%. Patients who received an antipseudomonal beta-lactam antibiotic with or without an aminoglycoside had a significantly higher cure rate than patients who received only an aminoglycoside (72% and 71% vs 29%). Patients with shock, pneumonia, or persistent neutropenia had a substantially poorer prognosis.


[Pathological classification of malignant peritoneal mesothelioma and comparative analysis with peritoneal carcinomatosis].
Objective: To analyze the pathological classification of malignant peritoneal mesothelioma (MPeM) and screen the immunohistochemical markers that can distinguish MPeM from peritoneal metastatic carcinoma (PC) . Methods: In June 2020, the pathological results of peritoneal biopsy of 158 MPeM and 138 PC patients from Cangzhou Central Hospital, Cangzhou People's Hospital, and Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine from May 2011 to July 2019 were retrospectively analyzed, and the pathological classifications of MPeM in Cangzhou were summarized. Immunohistochemical markers of MPeM and PC patients were analyzed, and receiver operating characteristic curve (ROC curve) was drawn for differential diagnosis of MPeM and PC. Results: There were 55 male and 103 female MPeM patients in Cangzhou, with an average age of 57.1 years old. The asbestos exposure rate was 91.14% (144/158). The most common pathological classifications were cutaneous type, accounting for 90.51% (143/158). There were significant differences in the expression of calreticulum protein, CK5/6, vimentin, D2-40, carcinoembryonic antigen (CEA) and tail type homologous nuclear gene transcription factor 2 (CDX-2) between MPeM and PC (P<0.05). Among the 6 positive markers, the sensitivity of calreticulum protein was the highest (0.905) and CEA was the lowest (0.428) . Conclusion: Calreticulum protein, CK5/6, vimentin, D2-40, CEA and CDX-2 may be used as specific markers to distinguish the diagnosis of MPeM from PC.


TLR7 ligands induce higher IFN-alpha production in females.
IFN-alpha exercises multiple immune modulatory and antiviral activities and has been suggested to play a critical role in the pathogenesis of systemic lupus erythematosus (SLE). Plasmacytoid dendritic cells (pDCs) release IFN-alpha upon TLR7 and TLR9 ligation. With respect to the nine times higher incidence of SLE in women and the clinical use of synthetic TLR ligands as novel immune adjuvants, we analyzed IFN-alpha and TNF-alpha production in healthy human individuals. Blood samples were incubated with synthetic TLR7 and TLR9 ligands. In three independent groups (n(1) = 120, n(2) = 101, and n(3) = 123), analysis revealed a capacity of female PBLs to produce significantly higher IFN-alpha levels after TLR7 stimulation (p(1) < 0.0000001, p(2) < 0.0000001, and p(3) < 0.0001) compared with male PBLs. In contrast, no sex differences were evident after TLR9 stimulation. TNF-alpha production after TLR7 stimulation and also total pDC numbers were not different between females and males. X-inactivation escape of the TLR7 gene was investigated in monoclonal B cell lines and, independently, in pDCs after cell sorting and single-cell picking, indicating regular silencing of one TLR7 allele in females. Additionally, exogenous 17beta-estrogen and estrogen receptor antagonism did not indicate a significant role on TLR7-induced IFN-alpha production. Our data reveal for the first time a profound sex-dependent pathway of TLR7-induced IFN-alpha with higher production in females. These findings may explain the higher prevalence of SLE in females and the reported decreased therapeutic efficacy of synthetic TLR7 ligands in male individuals.


Involvement of the leaf antioxidant system in the response to soil flooding in two Trifolium genotypes differing in their tolerance to waterlogging.
A comparative study of the response to waterlogging in a tolerant (Trifolium repens L., white clover cultivar Rivendel) and susceptible (Trifolium pratense L., red clover cultivar Raya) plants was undertaken to reveal the possible link between plant performance and oxidative stress protection mechanisms in leaves. Two weeks of soil waterlogging induced visible leaf damage in the susceptible genotype. In the tolerant one, signs of stress suffering appeared a week later. Waterlogging induced hydrogen peroxide accumulation in both clover species. The content of lipid hydroperoxides markedly increased in the sensitive plants along with stress prolongation, while in the tolerant ones their initial rise was followed by return to control levels. In the leaves of both genotypes ascorbic acid content increased following treatment, accompanied by transient increase in oxidized ascorbate. Superoxide dismutase (SOD) isoforms responded differently to the treatment, CuZn SOD isoforms being inhibited; catalase activity diminished while peroxidase activity increased and a new peroxidase isoform was detected after prolonged waterlogging in both clover species. Results support more pronounced oxidative secondary stress in red clover leaves as a result of waterlogging with progressively increased oxidative membrane injury, protein loss, and peroxidase activity enhancement. White clover presented relative protein stability and earlier and more active ascorbate involvement in the antioxidative protection.


Current status of veterinary vaccines.
The major goals of veterinary vaccines are to improve the health and welfare of companion animals, increase production of livestock in a cost-effective manner, and prevent animal-to-human transmission from both domestic animals and wildlife. These diverse aims have led to different approaches to the development of veterinary vaccines from crude but effective whole-pathogen preparations to molecularly defined subunit vaccines, genetically engineered organisms or chimeras, vectored antigen formulations, and naked DNA injections. The final successful outcome of vaccine research and development is the generation of a product that will be available in the marketplace or that will be used in the field to achieve desired outcomes. As detailed in this review, successful veterinary vaccines have been produced against viral, bacterial, protozoal, and multicellular pathogens, which in many ways have led the field in the application and adaptation of novel technologies. These veterinary vaccines have had, and continue to have, a major impact not only on animal health and production but also on human health through increasing safe food supplies and preventing animal-to-human transmission of infectious diseases. The continued interaction between animals and human researchers and health professionals will be of major importance for adapting new technologies, providing animal models of disease, and confronting new and emerging infectious diseases.


Comparison of accidental pediatric scald burns in a tertiary care center: hot cauldron burns versus accidental spill burns.
Background: Scald burns result from exposure to high-temperature fluids and are more common in the pediatric age group. They occur mainly by two mechanisms: (i) spill and (ii) immersion (hot cauldron) burns. These two patterns differ in clinical characteristics and outcomes. Scalds cause significant morbidity and mortality in children. The objective of this study was to compare accidental spill burns and hot cauldron burns in a hospital setting. Methods: An analytical cross-sectional study was conducted by reviewing the secondary data of scald cases admitted during the years 2019 and 2020 in a burn-dedicated tertiary care center. Total population sampling was adopted. Data analysis was done partly using SPSS, version-23, and Stata-15. Mann Whitney U-test and Chi-square/Fisher's exact test were done appropriately to find associations between different variables. Binary regression analysis was performed taking mortality events as the outcome of interest. Results: Out of 108 scald cases, 43 (39.8%) had hot cauldron burns and 65 (60.2%) had accidental spill burns. Overall mortality was 16 (14.8%), out of which hot cauldron burns and accidental spill burns comprised 12 (75.0%) and 4 (25.0%), respectively. Binary logistic regression analysis showed the type of scald, age, and Baux score found to be associated with mortality. Every one-year increment in age had a 29% lower odds of occurrence of mortality event (adjusted odds ratio [OR], 0.71; 95% confidence interval [CI], 0.50-0.99, p=0.042). Likewise, every one-point increment in Baux score was associated with 19% higher odds of mortality (adjusted OR, 1.190; 95% CI, 1.08-1.32; p<0.001). Conclusions: Accidental spill burn was more common but mortality was significantly higher for hot cauldron burns. The risk of mortality was significantly higher in burn events occurring outside the house, and burns involving back, buttocks, perineum, and lower extremities.


The watershed: a factor in coronary vein graft occlusion.
In 50 patients with one or more aortocoronary saphenous vein grafts investigated by angiography the patency rate of grafts inserted into arteries with a total proximal occlusion was significantly higher than of those inserted into arteries with a proximal stenosis. The interval between operation and investigation in the two groups was similar. In 10 patients with double grafts, one to an occluded and one to a non-occluded artery, there were 7 with one blocked graft, in each case to the non-occluded artery. In the other 3 both grafts were patent. A watershed was shown at angiography in one patient with a graft inserted into a stenosed artery; the distal run-off appeared good, but reflux of the contrast up the coronary artery into the aorta occurred when injecting into the graft, and vice versa. This watershed may operate to a minor degree in all grafts inserted into non-occluded arteries, and by causing stasis at the anastomosis, could explain the higher incidence of graft occlusion in this group. Ligation of the coronary artery proximal to the anastomosis may therefore be necessary to achieve the highest patency rate.


Swept-Source OCT Angiographic Characteristics of Treatment-Naïve Nonexudative Macular Neovascularization in AMD Prior to Exudation.
To investigate the characteristics of treatment-naïve nonexudative macular neovascularization (MNV) in age-related macular degeneration before the onset of exudation using swept-source optical coherence tomography angiography. MNV area, choriocapillaris (CC) flow deficits (FDs), vessel area density (VAD), vessel skeleton density (VSD), retinal pigment epithelial detachment (PED) volume, mean choroidal thickness (MCT), and choroid vascularity index (CVI) measurements were assessed at two visits prior to exudation. We compared measurements made at the second visit and the rate of change between visits in eyes with and without exudation. The differences in these parameters between eyes with and without subsequent exudation were summarized with 95% confidence intervals (CIs). Twenty-one eyes with nonexudative MNV were identified and followed. Nine eyes developed exudation, and 12 eyes did not develop exudation. Differences between these groups of eyes for all parameters tended to be small, and the 95% CIs largely ruled out any substantial differences. Overall, eyes with exudation had 24% smaller VAD, 20% smaller VSD, and 33% smaller PED volume measurements. No noteworthy differences were observed for MNV area, CC FDs, MCT, or CVI measurements. The onset of exudation was correlated with lesions having less vascularity and smaller PED volume measurements, but measurements of MNV area, CC FDs, MCT, and CVI were not correlated with near-term exudation. Investigations are ongoing to further explore these and other anatomic changes as harbingers of near-term exudation.


The role of physical activity in early adulthood and middle-age on bone health after menopause in epidemiological population from Silesia Osteo Active Study.
Osteoporosis is a growing problem in women after menopause. Among factors protecting from this disease is a physical activity (PA). The objective of the study was to evaluate the influence of PA in early adulthood and at middle-age on bone health after menopause. The study group consisted of 362 randomly recruited postmenopausal women after menopause. Mean age was 65.2±6.9 years. Medical history was collected from all participants, and they completed questionnaires assessing PA in early adulthood and at middle-age. Physical capacity was estimated using the Duke scale. Bone status was measured with use of densitometry (DXA) for lumbar spine and hip and calcaneus quantitative ultrasound (QUS) measurements. Physical activity and healthy lifestyle activity in early adulthood did not correlate with bone health, but current PA did (for QUS parameters r=.11; P<.05). Physical capacity correlated with all QUS parameters (r=.2 to .22; P<.05) and femoral neck (FN) BMD and T-score (for both r=.16; P<.05). Current PA frequency at the level of several times a week has the highest positive impact on FN DXA results (P=.01). Bone mass in DXA and QUS variables is related to the type of exercises (P<.05). Physical activity, especially several times a week, at middle-age is a major factor influencing bone health in women after menopause. Further studies concerning the type of exercises are needed.


MicroRNAs/TP53 feedback circuitry in glioblastoma multiforme.
MicroRNAs (miRNAs) are increasingly implicated in regulating cancer initiation and progression. In this study, two miRNAs, miR-25 and -32, are identified as p53-repressed miRNAs by p53-dependent negative regulation of their transcriptional regulators, E2F1 and MYC. However, miR-25 and -32 result in p53 accumulation by directly targeting Mdm2 and TSC1, which are negative regulators of p53 and the mTOR (mammalian target of rapamycin) pathway, respectively, leading to inhibition of cellular proliferation through cell cycle arrest. Thus, there is a recurrent autoregulatory circuit involving expression of p53, E2F1, and MYC to regulate the expression of miR-25 and -32, which are miRNAs that, in turn, control p53 accumulation. Significantly, overexpression of transfected miR-25 and -32 in glioblastoma multiforme cells inhibited growth of the glioblastoma multiforme cells in mouse brain in vivo. The results define miR-25 and -32 as positive regulators of p53, underscoring their role in tumorigenesis in glioblastoma.


Blood sampling through peripheral venous catheters is reliable for selected basic analytes in children.
The goal was to determine the interchangeability of peripheral venous catheter (PVC) and venipuncture blood sampling (BS). Paired blood samples from hospitalized children were obtained through venipuncture and from existing PVCs, following discard of 2 mL of blood. Comparisons of 9 complete blood count indices (white and red blood cell counts, hemoglobin and hematocrit levels, mean corpuscular volume, mean corpuscular hemoglobin level, red blood cell distribution width, platelet count, and mean platelet volume) and 5 basic chemical analysis indices (sodium, potassium, glucose, chloride, and urea levels) were performed, and hemolysis was documented. Irrespective of gauge, blood samples were obtained successfully from 40 (85.1%) of 47 PVCs, with no abnormal hemolysis. BS through venipuncture took longer than BS from PVCs (175.8 +/- 229.6 vs 104.5 +/- 53.4 seconds; P = .053) and was associated with significantly more distress/crying (73.1% vs 0%; P < .001). There were no significant differences between venipuncture and PVC samples (paired t test). Twenty-one (6%) of 348 pairs analyzed with the Clinical Laboratory Improvement Amendment standards fell outside the range of acceptable variance (8 of 21 aberrations were attributed to glucose measurements). Bland-Altman analysis indicated that, with the exclusion of glucose measurements, BS from PVCs is reliable, with 29 (6.5%) of 448 pairs exceeding the limits of agreement. Of those, 9 cases were clinically significant, but none would have altered clinical management. PVC sampling was shown to be a pain-reducing method that can be used for children for selected basic analytes. The findings for glucose were unreliable.


[Effects of crocin on hippocampus rapid kindling epilepsy in mice].
Objective: To investigate the effect of crocin on the progression and generalized seizure of temporal lobe epilepsy in mice. Methods: Hippocampus rapid kindling model was established in C57BL/6J mice. The effects of crocin on seizure stage, afterdischarge duration (ADD), number of stimulation in each stage and final state, the incidence of generalized seizure (GS), average seizure stage and ADD were observed. Results: Crocin (20 mg/kg) significantly retarded behavioral seizure stages ( P<0.05) and shortened cumulative ADD ( P<0.01) during hippocampus rapid kindling acquisition in mice compared with vehicle group. Meanwhile, number of stimulations in stage 1-2 was significantly increased ( P<0.05) and the incidence of fully kindled animals was significantly decreased ( P<0.01). However, 10 or 50 mg/kg crocin showed no significant effect on the above indexes (all P>0.05). Crocin (100 or 200 mg/kg) significantly decreased the incidence of GS (all P<0.01) and reduced average seizure stages (all P<0.01) in fully-kindled mice compared with vehicle group; Fifty mg/kg crocin only reduced average seizure stages ( P<0.05). Conclusion: Low-dose crocin can retard the progression in hippocampus rapid kindling acquisition in mice, while high-dose crocin relieves the GS in fully-kindled mice, which suggests that crocin may be a potential anti-epileptic compound.


A unique serotonin receptor in choroid plexus is linked to phosphatidylinositol turnover.
A novel serotonergic binding site, the 5-HT1C site, has been characterized recently in choroid plexus and several brain regions. The biochemical and physiological roles of this site have not been previously described. In this report we show that serotonin (5-hydroxytryptamine, 5-HT) stimulates phosphatidylinositol turnover in rat choroid plexus. The pharmacology of serotonin-stimulated phosphatidylinositol hydrolysis in choroid plexus was compared to the pharmacology in cerebral cortex, where this response is mediated by the serotonin 5-HT2 receptor. Serotonin increased phosphatidylinositol turnover in choroid plexus by 6-fold and in cerebral cortex by 2.5-fold. Serotonin was greater than 10-fold more potent in choroid plexus (EC50 = 46 nM) than in cerebral cortex (EC50 = 540 nM). The serotonin antagonists ketanserin, mianserin, and spiperone inhibited the response in the two tissues with different potencies. In cerebral cortex all three exhibited nanomolar affinities consistent with their potencies at the 5-HT2 site. In choroid plexus, however, the rank order (mianserin greater than ketanserin much greater than spiperone) and absolute potencies were consistent with binding to the 5-HT1C site. These data suggest that the 5-HT1C site in choroid plexus is a functional receptor that utilizes phosphatidylinositol turnover as its biochemical effector system.


Site-specific cleavage of the host poly(A) binding protein by the encephalomyocarditis virus 3C proteinase stimulates viral replication.
Although picornavirus RNA genomes contain a 3'-terminal poly(A) tract that is critical for their replication, the impact of encephalomyocarditis virus (EMCV) infection on the host poly(A)-binding protein (PABP) remains unknown. Here, we establish that EMCV infection stimulates site-specific PABP proteolysis, resulting in accumulation of a 45-kDa N-terminal PABP fragment in virus-infected cells. Expression of a functional EMCV 3C proteinase was necessary and sufficient to stimulate PABP cleavage in uninfected cells, and bacterially expressed 3C cleaved recombinant PABP in vitro in the absence of any virus-encoded or eukaryotic cellular cofactors. N-terminal sequencing of the resulting C-terminal PABP fragment identified a 3C(pro) cleavage site on PABP between amino acids Q437 and G438, severing the C-terminal protein-interacting domain from the N-terminal RNA binding fragment. Single amino acid substitution mutants with changes at Q437 were resistant to 3C(pro) cleavage in vitro and in vivo, validating that this is the sole detectable PABP cleavage site. Finally, while ongoing protein synthesis was not detectably altered in EMCV-infected cells expressing a cleavage-resistant PABP variant, viral RNA synthesis and infectious virus production were both reduced. Together, these results establish that the EMCV 3C proteinase mediates site-specific PABP cleavage and demonstrate that PABP cleavage by 3C regulates EMCV replication.


Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit.
Genome editing facilitated by Cas9-based RNA-guided nucleases (RGNs) is becoming an increasingly important and popular technique for reverse genetics in both model and non-model species. So far, RGNs were mainly applied for the induction of point mutations, and one major challenge consists in the detection of genome-edited individuals from a mutagenized population. Also, point mutations are not appropriate for functional dissection of non-coding DNA. Here, the multiplexing capacity of a newly developed genome editing toolkit was exploited for the induction of inheritable chromosomal deletions at six different loci in Nicotiana benthamiana and Arabidopsis. In both species, the preferential formation of small deletions was observed, suggesting reduced efficiency with increasing deletion size. Importantly, small deletions (<100 bp) were detected at high frequencies in N. benthamiana T0 and Arabidopsis T2 populations. Thus, targeting of small deletions by paired nucleases represents a simple approach for the generation of mutant alleles segregating as size polymorphisms in subsequent generations. Phenotypically selected deletions of up to 120 kb occurred at low frequencies in Arabidopsis, suggesting larger population sizes for the discovery of valuable alleles from addressing gene clusters or non-coding DNA for deletion by programmable nucleases.


Combined doppler and phased-array echocardiographic estimation of cardiac output.
The capability of a pulsed Doppler flowmeter combined with a phased-array imaging system to measure volume flow was tested in vitro and in patients undergoing cardiac catheterization. The Doppler-phased-array system (DPA) was used to determine vessel diameter and a superimposed cursor was used to locate the range and angle of the Doppler sample volume. DPA estimates of continuous flow through tubing in a water tank correlated strongly (r = 0.99) with measured flow corresponding to physiologic ranges from 3-12 l/min. For pulsatile flow in a water tank, a correlation of r = 0.86 with measured flow was obtained, whereas DPA estimates of cardiac outputs compared with Fick estimates in the 11 patients produced a correlation of r = 0.83. These data indicate that estimates of cardiac output are possible using the DPA approach.


[Investigation into vegetative regulation of blood circulation during 7-day dry immersion].
The paper presents the results of investigations into vegetative regulation of blood circulation and regulation modification by peroral amlodipine and myostimulation during 7-d dry immersion. It was shown that in immersion vegetative regulation readjusted towards predominance of the sympathetic mechanisms. Myostimulation and peroral amlodipine modified regulation substantially mobilizing high level suprasegmentary structures. It should be noted that amlodipine and myostimulation differ in mechanisms of their action on central regulation. Pharmaceutical intervention seems to have a more complex and varying effect on people including side-effects. Presumably, it was the cause of poor orthostatic tolerance of several test-subjects.


Maternal Fat Feeding Augments Offspring Nephron Endowment in Mice.
Increasing consumption of a high fat 'Western' diet has led to a growing number of pregnancies complicated by maternal obesity. Maternal overnutrition and obesity have health implications for offspring, yet little is known about their effects on offspring kidney development and renal function. Female C57Bl6 mice were fed a high fat diet (HFD, 21% fat) or matched normal fat diet (NFD, 6% fat) for 6 weeks prior to pregnancy and throughout gestation and lactation. HFD dams were overweight and glucose intolerant prior to mating but not in late gestation. Offspring of NFD and HFD dams had similar body weights at embryonic day (E)15.5, E18.5 and at postnatal day (PN)21. HFD offspring had normal ureteric tree development and nephron number at E15.5. However, using unbiased stereology, kidneys of HFD offspring were found to have 20-25% more nephrons than offspring of NFD dams at E18.5 and PN21. Offspring of HFD dams with body weight and glucose profiles similar to NFD dams prior to pregnancy also had an elevated nephron endowment. At 9 months of age, adult offspring of HFD dams displayed mild fasting hyperglycaemia but similar body weights to NFD offspring. Renal function and morphology, measured by transcutaneous clearance of FITC-sinistrin and stereology respectively, were normal. This study demonstrates that maternal fat feeding augments offspring nephron endowment with no long-term consequences for offspring renal health. Future studies assessing the effects of a chronic stressor on adult mice with augmented nephron number are warranted, as are studies investigating the molecular mechanisms that result in high nephron endowment.


First Demonstration of Electrostatic Damping of Parametric Instability at Advanced LIGO.
Interferometric gravitational wave detectors operate with high optical power in their arms in order to achieve high shot-noise limited strain sensitivity. A significant limitation to increasing the optical power is the phenomenon of three-mode parametric instabilities, in which the laser field in the arm cavities is scattered into higher-order optical modes by acoustic modes of the cavity mirrors. The optical modes can further drive the acoustic modes via radiation pressure, potentially producing an exponential buildup. One proposed technique to stabilize parametric instability is active damping of acoustic modes. We report here the first demonstration of damping a parametrically unstable mode using active feedback forces on the cavity mirror. A 15 538 Hz mode that grew exponentially with a time constant of 182 sec was damped using electrostatic actuation, with a resulting decay time constant of 23 sec. An average control force of 0.03 nN was required to maintain the acoustic mode at its minimum amplitude.


Vaccination concerns, beliefs and practices among Ukrainian migrants in Poland: a qualitative study.
Ukrainians numbering approximately 1.2 million are the largest migrant group in Poland. Data on vaccination coverage among migrants are not collected in EU, including Poland. Therefore, this qualitative study aimed to identify vaccination practices in this migrant group, to explore facilitators and barriers to vaccination and related access to Polish healthcare services. In September 2019, a qualitative study of Ukrainian migrants (UMs) living in Szczecin, Poland, and recruited through a snowball sampling method, was conducted. Using a semi-structured topic guide, four focus groups were held with 22 UMs aged 18-45. Participants were asked about their attitudes towards vaccination in general with comparison between services in Poland and Ukraine. Following transcription and translation, a thematic analysis was conducted. Respondents were distrustful of Ukrainian vaccination policy, medical personnel and individual vaccines, however, they often returned to Ukraine for dental and gynaecological appoint-ments. While critical with regards to registering with Polish GPs practices, UMs were confident in health professionals, as well as vaccine delivery. Vaccines were perceived as safer and of better quality than in Ukraine. Difficulties in translating vaccination records were rarely reported, verbal communi-cation was not problematic due to language similarities. All UM parents reported vaccinating their children according to the Polish schedule. However, a significant number of adult UMs have not completed mandatory vaccinations, although they may have obtained false immunization certificates; according to UMs those can be obtained by bribing. Participants reported lower acceptance of the influenza vaccine, mainly due to perceptions around its importance; none had been vaccinated against influenza. None of UMs had heard of the HPV vaccine. UMs experienced challenges in accessing credible online vaccination information in Ukrainian, no official local health authority vaccination material existed either, except for information about measles. This study pinpointed positive UM attitudes and practices regarding child vaccination in the Polish healthcare system and identified issues for improvement, such as adult vaccination. Health communication should be more tailored within UMs information delivery systems to enable migrants to make informed choices about vaccination. Further research is needed to better assess factors affecting vaccine uptake identified in this study.