Receptor-binding properties in vitro and in vivo of ritanserin: A very potent and long acting serotonin-S2 antagonist.
In vitro and in vivo receptor-binding properties of the new serotonin antagonist, ritanserin, are reported. In in vitro binding assays, ritanserin shows high affinity binding to serotonin-S2 sites in rat frontal cortex tissue: IC50 = 0.9 nM without drug preincubation and 0.3 nM with 30-min drug preincubation; IC50 values for histamine-H1, dopamine-D2, and adrenergic-alpha 1 and -alpha 2 sites were 39-, 77-, 107-, and 166-fold higher, and at up to 1 microM, the drug did not bind to serotonin-S1 sites. In in vitro assays, ritanserin dissociated very slowly from serotonin-S2 (t1/2 = 160 min) and histamine-H1 sites (t1/2 = 77 min) and rapidly from dopamine-D2 sites (t1/2 = 11 min). Half-times of dissociation from adrenergic-alpha 1 and -alpha 2 sites were 18 and 26 min. The inhibition by ritanserin of [3H]ketanserin binding was found to be partially noncompetitive and the inhibitory potency increased with drug preincubation. Due to the slow dissociation of ritanserin from the serotonin-S2 sites, the drug cannot be displaced completely by [3H]ketanserin. In contrast, inhibition by ritanserin of [3H]haloperidol binding to dopamine-D2 sites in rat striatum was fully competitive, in agreement with the rapid dissociation of the drug from the latter sites. In ex vivo binding assays using brain areas of rats and guinea pigs treated subcutaneously with ritanserin, occupation of serotonin-S2 sites was observed at very low dosage (50% occupation at 0.08-0.1 mg/kg) and sites remained occupied during a prolonged time period (greater than 70% occupation up to 48 hr after 2.5 mg/kg ritanserin). Histamine-H1 receptor sites in guinea pig cerebellum became occupied at dosages 25-fold higher than the dosage producing occupation of frontal cortical serotonin-S2 sites. Dopamine-D2 sites in rat striatum and cortical adrenergic-alpha 1 sites became only slightly occupied (less than 20%) at higher dosages and the effect was not dose-dependent. Adrenergic-alpha 2 sites were not occupied up to doses of 160 mg/kg given subcutaneously. In vivo binding assays using [3H]spiperone confirmed the occupation of frontal cortical serotonin-S2 sites following low dosage of ritanserin and a minor occupation of striatal dopamine-D2 sites. Levels of dopamine and serotonin and their metabolites remained unchanged in brain areas of rats orally treated with ritanserin up to dosages of 40 mg/kg. At 160 mg/kg, there seemed to be a slight reduction in dopamine and serotonin content.(ABSTRACT TRUNCATED AT 400 WORDS)


Do prostaglandins have a role in breathlessness?
The effects of indomethacin on the relationship between breathlessness and minute ventilation during exercise have been determined in a double-blind, randomized study on 6 normal subjects. Indomethacin did not significantly after ventilation or oxygen uptake either at rest or during submaximal exercise. Breathlessness was assessed with visual analogue scales, and, when compared with placebo, the sensation was significantly reduced in relation to ventilation (p less than 0.02). These findings provide evidence of a possible role for prostanoids in the mechanisms that give rise to breathlessness.


Differential lectin binding patterns in the oviductal ampulla of the horse during oestrus.
We investigated the oligosaccharide sequence of glycoconjugates, mainly sialoglycoconjugates, in the horse oviductal ampulla during oestrus by means of lectin and pre-lectin methods such as the KOH-neuraminidase procedure to remove sialic acid residues and incubation with N-glycosidase F to cleave N-linked glycans. Ciliated cells displayed N-linked oligosaccharides throughout the cytoplasm. The cilia glycocalyx expressed both N- and O-linked (mucin-type) oligosaccharides, both showing a high variety of terminal sequences. In the most non-ciliated cells, the whole cytoplasm contained N-linked oligosaccharides with terminal alphaGal as well as mucin-type glycans with terminal Forssman pentasaccharides. In a few scattered non-ciliated cells, the whole cytoplasm displayed sialylated N-linked oligosaccharides with terminal Neu5Ac-GalNAc and O-linked glycans terminating with neutral and/or alphaGalNAc, Neu5Ac alpha2,6Gal/GalNAc, Neu5AcGal beta1,3GalNAc. Supra-nuclear granules, probably Golgi zones, of non-ciliated cells showed mainly O-linked glycans rich in sialic acid residues. The luminal surface of non-ciliated cells showed N-linked oligosaccharides, containing terminal/internal alphaMan/alphaGlc, betaGlcNAc and terminal alphaGal, as well as mucin-type oligosaccharides terminating with a large variety of either neutral saccharides or sialylated sequences. Apical protrusions containing O-linked oligosaccharides with terminal Forssman pentasaccharide, Neu5Ac-Gal beta1,4GlcNAc, Neu5Ac-GalNAc were seen in non-ciliated cells scattered along the epithelium. These findings show the presence of sialoglycoconjugates in the oviductal ampulla epithelium of the mare and the existence of different lectin binding profiles between ciliated and non-ciliated (secretory) cells, as well as the presence of non-ciliated cell sub-types which might determine functional differences along the ampullary epithelium of mare oviduct.


Fast and accurate quantification of insertion-site specific transgene levels from raw seed samples using solid-state nanopore technology.
Many modern crop varieties contain patented biotechnology traits, and an increasing number of these crops have multiple (stacked) traits. Fast and accurate determination of transgene levels is advantageous for a variety of use cases across the food, feed and fuel value chain. With the growing number of new transgenic crops, any technology used to quantify them should have robust assays that are simple to design and optimize, thereby facilitating the addition of new traits to an assay. Here we describe a PCR-based method that is simple to design, starts from whole seeds, and can be run to end-point in less than 5 minutes. Subsequent relative quantification (trait vs. non-trait) using capillary electrophoresis performed in 5% increments across the 0-100% range showed a mean absolute error of 1.9% (s.d. = 1.1%). We also show that the PCR assay can be coupled to non-optical solid-state nanopore sensors to give seed-to-trait quantification results with a mean absolute error of 2.3% (s.d. = 1.6%). In concert, the fast PCR and nanopore sensing stages demonstrated here can be fully integrated to produce seed-to-trait quantification results in less than 10 minutes, with high accuracy across the full dynamic range.


VRK2 inhibits mitogen-activated protein kinase signaling and inversely correlates with ErbB2 in human breast cancer.
The epidermal growth factor (EGF)-ErbB-mitogen-activated protein kinase (MAPK) transcription signaling pathway is altered in many types of carcinomas, and this pathway can be regulated by new protein-protein interactions. Vaccinia-related kinase (VRK) proteins are Ser-Thr kinases that regulate several signal transduction pathways. In this work, we study the effect of VRK2 on MAPK signaling using breast cancer as a model. High levels of VRK2 inhibit EGF and ErbB2 activation of transcription by the serum response element (SRE). This effect is also detected in response to H-Ras(G12V) or B-Raf(V600E) oncogenes and is accompanied by a reduction in phosphorylated extracellular signal-regulated kinase (ERK) levels, p90RSK levels, and SRE-dependent transcription. Furthermore, VRK2 knockdown has the opposite effect, increasing the transcriptional response to stimulation with EGF and leading to increased levels of ERK phosphorylation. The molecular mechanism lies between MAPK/ERK kinase (MEK) and ERK, since MEK remains phosphorylated while ERK phosphorylation is blocked by VRK2A. This inhibition of the ERK signaling pathway is a consequence of a direct protein-protein interaction between VRK2A, MEK, and kinase suppressor of Ras 1 (KSR1). Identification of new correlations in human cancer can lead to a better understanding of the biology of individual tumors. ErbB2 and VRK2 protein levels were inversely correlated in 136 cases of human breast carcinoma. In ErbB2(+) tumors, there is a significant reduction in the VRK2 level, suggesting a role for VRK2A in ErbB2-MAPK signaling. Thus, VRK2 downregulation in carcinomas permits signal transmission through the MEK-ERK pathway without affecting AKT signaling, causing a signal imbalance among pathways that contributes to the phenotype of breast cancer.


A developmental theory of synaesthesia, with long historical roots: a comment on Hochel & Milan.
The recent surge of scientific investigation into synaesthesia, ably reviewed by Hochel and Milan (2008), is representative of an increasing recognition that our various sensory modalities are intimately interconnected rather than separate. The origin of these interconnections is the subject of an intriguing theory by Maurer and Maurer (1988). They suggest that all of us begin life as synaesthetes, with subsequent neural development reducing the connections among the senses. We present some historical roots of the idea that human life begins with the senses intertwined. The influential 18th-century philosopher Jean-Jacques Rousseau described an early theory of child development in his book Emile (1762), hypothesizing that if "a child had at its birth the stature and strength of a man ... all his sensations would be united in one place, they would exist only in the common 'sensorium'." A half-century later, a young Mary Shelley (1818) brought this idea into popular culture with the Frankenstein creature's recollection of his early experience: "A strange multiplicity of sensations seized me, and I saw, felt, heard, and smelt, at the same time; and it was, indeed, a long time before I learned to distinguish between the operations of my various senses." William James in The Principles of Psychology (1890) expressed a similar idea. In this context, the assumption of many 20th-century scientists that the senses were largely separate appears to be an historical aberration.


Revisiting an ecophysiological oddity: Hydathode-mediated foliar water uptake in Crassula species from southern Africa.
Hydathodes are usually associated with water exudation in plants. However, foliar water uptake (FWU) through the hydathodes has long been suspected in the leaf-succulent genus Crassula (Crassulaceae), a highly diverse group in southern Africa, and, to our knowledge, no empirical observations exist in the literature that unequivocally link FWU to hydathodes in this genus. FWU is expected to be particularly beneficial on the arid western side of southern Africa, where up to 50% of Crassula species occur and where periodically high air humidity leads to fog and/or dew formation. To investigate if hydathode-mediated FWU is operational in different Crassula species, we used the apoplastic fluorescent tracer Lucifer Yellow in combination with different imaging techniques. Our images of dye-treated leaves confirm that hydathode-mediated FWU does indeed occur in Crassula and that it might be widespread across the genus. Hydathodes in Crassula serve as moisture-harvesting structures, besides their more common purpose of guttation, an adaptation that has likely played an important role in the evolutionary history of the genus. Our observations suggest that ability for FWU is independent of geographical distribution and not restricted to arid environments under fog influence, as FWU is also operational in Crassula species from the rather humid eastern side of southern Africa. Our observations point towards no apparent link between FWU ability and overall leaf surface wettability in Crassula. Instead, the hierarchically sculptured leaf surfaces of several Crassula species may facilitate FWU due to hydrophilic leaf surface microdomains, even in seemingly hydrophobic species. Overall, these results confirm the ecophysiological relevance of hydathode-mediated FWU in Crassula and reassert the importance of atmospheric humidity for some arid-adapted plant groups.


Decreased urea synthesis in cafeteria-diet-induced obesity in the rat.
Feeding rats with a cafeteria diet resulted in increases in total body weight and in epididymal-adipose-tissue weight. Those rats excreted significantly less N than did controls. The amount of N ingested by cafeteria-diet-fed rats was kept equal to that of controls. This decrease in N excretion is explained by a decrease in urinary excretion of urea. This may be due to the following facts. The rate of synthesis of urea from precursors by isolated hepatocytes from cafeteria-diet-fed rats was lower than in controls. In cafeteria-diet-fed rats the activities of all the enzymes of the urea cycle are decreased. The major percentage decreases are those of carbamoylphosphate synthetase (EC 220.127.116.11) and of argininosuccinate synthetase (EC 188.8.131.52), the enzymes probably involved in the regulation of the overall rate of the cycle. When rats are switched to normal chow diet, the enzyme activities return to normal values. The uptake of amino acids by liver of cafeteria-diet-fed rats is lower than in controls. These results contrast with those obtained previously by using other models of obesity in rat (i.e. genetic or hypothalamic), in which N excretion was increased.


Modifying Epoxy Resins to Resist Both Fire and Water.
In this work, we converted inherently hydrophilic and flammable epoxy resins to resist both fire and water. This was achieved by integrating surface modified flame-retardant CoFe2O4 nanoparticles into the epoxy resin. The modified CoFe2O4 nanoparticles reduce the peak heat release rate, peak smoke production release and CO production in the CoFe2O4/epoxy resin nanocomposite by 39.6, 41.6, and 61.3%, respectively. The obtained flame retardant CoFe2O4/EP items demonstrate excellent durability with constant superhydrophobicity even after high external pressure, knife-scratch, and mechanical abrasion. In addition, the obtained items demonstrate outstanding robust water-repellent properties after facing long-term exposure to extremely corrosive liquids and also show self-cleaning properties in air and under oil.


CD83 is preformed inside monocytes, macrophages and dendritic cells, but it is only stably expressed on activated dendritic cells.
Human DCs (dendritic cells) express surface CD83 upon activation. Comparing the surface induction of CD83 with the upregulation of CD40, CD80 and CD86 during LPS (lipopolysaccharide)-induced DC maturation showed that CD83 induction occurred more rapidly. Despite the lack of CD83 on immature DCs, it was detected in these cells by Western blotting and flow cytometry. Indirect immunofluorescence revealed CD83 inside immature DCs in perinuclear regions. CD83 was absent on monocytes and macrophages, but it was detected inside these cells and found to be rapidly surface-expressed upon LPS-induced activation. Whereas CD83 expression on activated DCs was sustainable, its expression on monocytes and macrophages was transient. Optimal interleukin-4 co-stimulation during DC generation from monocytes was found to be essential for stable CD83 surface expression. CD83 was detected as 37 and 50 kDa forms in transfected 293T cells. Macrophages and immature DCs expressed the 37 kDa form, whereas mature DCs predominantly expressed the 50 kDa form. In monocytes, CD83 was detected as a 22 kDa detergent-insoluble form. The rapid CD83 surface induction on DCs and macrophages was blocked by brefeldin A, but not by cycloheximide, showing that fresh CD83 synthesis was not essential. Tunicamycin inhibited the expression of the 50 and 37 kDa CD83 forms, and also blocked CD83 surface expression on DCs and macrophages. PNGase F (peptide N-glycosidase F) digestion reduced the 37 and 50 kDa CD83 forms to 28 kDa. In summary, monocytes, macrophages and immature DCs contain preformed intracellular CD83, and its rapid surface expression upon activation is post-translationally regulated in a process involving glycosylation.


Antenatal depression across trimesters as a risk for postpartum depression and estimation of the fraction of postpartum depression attributable to antenatal depression: A systematic review and meta-analysis of cohort studies.
This meta-analysis aimed at estimating the prevalence of postpartum depression (PPD) at different postpartum timepoints in women with antenatal depression (AD) in the three trimesters. We also examined the association between AD and PPD, and estimated the population attributable fraction of PPD to AD. This systematic review and meta-analysis identified cohort studies that determined the prevalence of PPD in women who had AD, and those that examined the association between AD and PPD from PubMed, Embase, MEDLINE, CINAHL and PsycINFO. Articles were appraised using the modified Newcastle Ottawa Scale and data were analyzed using Comprehensive Meta-Analysis. Eighty-eight (88) cohort studies with a combined sample size of 1,042,448 perinatal women contributed to the meta-analysis. About 37% pregnant women who had AD, later had PPD. Those with AD had four times higher odds of developing PPD (OR: 4.58; 95% CI = 3.52-5.96). The odds of having PPD were higher when AD was observed in the first or third trimester compared to the second trimester. About 12.8% of PPD cases were attributable to AD. The findings should inform future clinical guidelines on the screening, the frequency of screening, and follow-up care in maternal-mental health.


The association between depressive symptoms and acute exacerbations of COPD.
Depression is an important comorbidity for patients with chronic obstructive pulmonary disease (COPD). The association between depression and acute exacerbations of COPD is unknown. This study was designed to determine the frequency of COPD exacerbations in outpatients with and without depressive symptoms. In this retrospective cohort study, patients with a primary diagnosis of COPD were followed for 1 year after discharge from a pulmonary rehabilitation program and the frequency of exacerbations was recorded. Upon completion of the program, all patients were administered the Short-Form 36 Health Survey (SF-36), which contains a mental health domain. Patients were classified as having depressive symptoms based on their domain score, which was separately validated in a second population of patients. Of the 194 patients with COPD who completed the pulmonary rehabilitation program, 32 (16.5%) had depressive symptoms. There were no differences in terms of age, race, pack-years, forced expiratory volume in 1 second (FEV(1)), 6-minute walk distance, body mass index, use of supplemental oxygen, use of inhaled steroids, or the Charlson Comorbidity Index between patients with and without depressive symptoms. Patients with depressive symptoms had more exacerbations in the following year (1.91 vs. 1.36; p = 0.02), were 2.8 times more likely to have ever had an exacerbation (95% confidence interval (CI), 1.1-7.3; p = 0.03), and suffered a first exacerbation earlier (148 days compared with 266 days; p = 0.04) than nondepressed patients. COPD patients with depressive symptoms have a significantly higher risk for exacerbations. Early screening for depression in patients with COPD may help identify those patients at higher risk for subsequent exacerbations.


Apparent lisinopril overdose requiring hemodialysis.
A case of apparent overdose of angiotensin-converting-enzyme inhibitors requiring hemodialysis is reported. A 51-year-old white man (weight, 85 kg; height, 178 cm; body mass index, 28) with a history of hypertension, low back pain, and anxiety apparently took 27 lisinopril 10-mg tablets (3.18 mg/kg body weight) over a period of 3 or fewer days. The friend who brought him to the emergency department reported that the patient was hard to rouse and was speaking incoherently on the day of admission. Over the previous few days, the patient reportedly had visual hallucinations, incoherence, and inarticulate speech. Laboratory tests, electrocardiography, and computed tomography were performed. The patient was judged to have high-anion-gap metabolic acidosis, acute kidney injury, severe hyperkalemia, and rhabdomyolysis. He was given three doses of albuterol via a nebulizer, three doses of calcium gluconate 1 g i.v., two doses of sodium bicarbonate 100 meq i.v., two doses of sodium polystyrene sulfonate 30 g orally, three doses of insulin 10 units i.v., and three doses of dextrose 25 g (as 50% dextrose injection) i.v. He then underwent emergent hemodialysis and was admitted to the intensive care unit. The patient's confusion abated, kidney function improved, and acid-base and electrolyte imbalances resolved. The patient was discharged after 15 days. A man who had evidently taken an overdose of lisinopril had multiorgan dysfunction in the absence of hypotension. The abnormalities resolved after he was treated for acidosis and hyperkalemia and received hemodialysis to remove the lisinopril.


Predicting XAFS scattering path cumulants and XAFS spectra for metals (Cu, Ni, Fe, Ti, Au) using molecular dynamics simulations.
The ability of molecular dynamics (MD) simulations to support the analysis of X-ray absorption fine-structure (XAFS) data for metals is evaluated. The low-order cumulants (ΔR, σ(2), C3) for XAFS scattering paths are calculated for the metals Cu, Ni, Fe, Ti and Au at 300 K using 28 interatomic potentials of the embedded-atom method type. The MD cumulant predictions were evaluated within a cumulant expansion XAFS fitting model, using global (path-independent) scaling factors. Direct simulations of the corresponding XAFS spectra, χ(R), are also performed using MD configurational data in combination with the FEFF ab initio code. The cumulant scaling parameters compensate for differences between the real and effective scattering path distributions, and for any errors that might exist in the MD predictions and in the experimental data. The fitted value of ΔR is susceptible to experimental errors and inadvertent lattice thermal expansion in the simulation crystallites. The unadjusted predictions of σ(2) vary in accuracy, but do not show a consistent bias for any metal except Au, for which all potentials overestimate σ(2). The unadjusted C3 predictions produced by different potentials display only order-of-magnitude consistency. The accuracy of direct simulations of χ(R) for a given metal varies among the different potentials. For each of the metals Cu, Ni, Fe and Ti, one or more of the tested potentials was found to provide a reasonable simulation of χ(R). However, none of the potentials tested for Au was sufficiently accurate for this purpose.


Does night-time aircraft noise trigger mortality? A case-crossover study on 24 886 cardiovascular deaths.
It is unclear whether night-time noise events, including from aeroplanes, could trigger a cardiovascular death. In this study, we investigate the potential acute effects of aircraft noise on mortality and the specific role of different night-time exposure windows by means of a case-crossover study design. We selected 24 886 cases of death from cardiovascular disease (CVD) from the Swiss National Cohort around Zürich Airport between 2000 and 2015. For night-time deaths, exposure levels 2 h preceding death were significantly associated with mortality for all causes of CVD [OR = 1.44 (1.03-2.04) for the highest exposure group (LAeq > 50 dB vs. <20 dB)]. Most consistent associations were observed for ischaemic heart diseases, myocardial infarction, heart failure, and arrhythmia. Association were more pronounced for females (P = 0.02) and for people living in areas with low road and railway background noise (P = 0.01) and in buildings constructed before 1970 (P = 0.36). We calculated a population attributable fraction of 3% in our study population. Our findings suggest that night-time aircraft noise can trigger acute cardiovascular mortality. The association was similar to that previously observed for long-term aircraft noise exposure.


Health-promoting effects of Lactobacillus-fermented barley in weaned pigs challenged with Escherichia coli K88.
Fermented feeds are being considered as practical alternatives to antimicrobial growth promoters (AGP) supplemented in nursery pig diets. This study aimed to investigate health-promoting effects of fermented barley in weaned pigs challenged with Escherichia coli K88 +. A total of 37 piglets were weaned at 21 ± 1 day of age (6.41 ± 0.47 kg of BW) and assigned to either of the following five treatment groups: (1) unchallenged control (UCC; n = 7), (2) challenged control (CC; n = 7), (3) AGP (CC + 0.1% AGP; n = 7), (4) Ferm1 (challenged and fed homofermentative Lactobacillus plantarum (Homo)-fermented barley; n = 8) and (5) Ferm2 (challenged and fed heterofermentative L. buchneri (Hetero)-fermented barley; n = 8). The control diet included unfermented barley. Barley was fermented with either Homo or Hetero for 90 days under anaerobic conditions. On day 10, all pigs except those in UCC group were orally inoculated with E. coli K88 + (6 × 109 colony forming units/ml). The pre-planned orthogonal test was performed to compare (1) UCC and CC, (2) CC and AGP, (3) CC and Ferm1 + Ferm2, as well as (4) Ferm1 and Ferm2. Challenged control pigs showed shorter (P < 0.05) villus height (VH) in the duodenum and deeper (P < 0.05) crypt depth (CD) in the jejunum than UCC pigs. The AGP group had higher (P < 0.05) VH and lower (P < 0.05) IL-6 gene expression in the jejunum compared with CC group. Compared to CC, Ferm1 and Ferm2 had decreased (P < 0.05) CD in the duodenum, IL-6 gene expression in the jejunum and rectal temperature at 24 h post-challenge. Pigs fed fermented barley diets showed greater (P < 0.05) faecal abundance of Clostridium Cluster IV and Lactobacilli than those fed UCC diet. Ferm2-fed pigs showed lower (P < 0.05) concentrations of band cells, eosinophils and lymphocytes at 6, 24 and 48 h after challenge, respectively, and lower (P < 0.05) faecal abundance of Enterobacteriaceae 24 h after challenge than the Ferm1-fed pigs. In conclusion, the substitution of unfermented barley with fermented barley in a nursery diet showed similar results as those shown by AGP supplementation in terms of enhancing the intestinal morphology and modulating faecal microbiota composition, as well as down-regulating the pro-inflammatory cytokines; therefore, fermented barley can be a possible nutritional strategy for managing nursery pigs fed diets without in-feed AGP.


Filament formation by metabolic enzymes is a specific adaptation to an advanced state of cellular starvation.
One of the key questions in biology is how the metabolism of a cell responds to changes in the environment. In budding yeast, starvation causes a drop in intracellular pH, but the functional role of this pH change is not well understood. Here, we show that the enzyme glutamine synthetase (Gln1) forms filaments at low pH and that filament formation leads to enzymatic inactivation. Filament formation by Gln1 is a highly cooperative process, strongly dependent on macromolecular crowding, and involves back-to-back stacking of cylindrical homo-decamers into filaments that associate laterally to form higher order fibrils. Other metabolic enzymes also assemble into filaments at low pH. Hence, we propose that filament formation is a general mechanism to inactivate and store key metabolic enzymes during a state of advanced cellular starvation. These findings have broad implications for understanding the interplay between nutritional stress, the metabolism and the physical organization of a cell.


Gas6 Promotes Inflammatory (CCR2hiCX3CR1lo) Monocyte Recruitment in Venous Thrombosis.
Coagulation and inflammation are inter-related. Gas6 (growth arrest-specific 6) promotes venous thrombosis and participates to inflammation through endothelial-innate immune cell interactions. Innate immune cells can provide the initiating stimulus for venous thrombus development. We hypothesize that Gas6 promotes monocyte recruitment during venous thrombosis. Deep venous thrombosis was induced in wild-type and Gas6-deficient (-/-) mice using 5% FeCl3 and flow reduction in the inferior vena cava. Total monocyte depletion was achieved by injection of clodronate before deep venous thrombosis. Inflammatory monocytes were depleted using an anti-C-C chemokine receptor type 2 (CCR2) antibody. Similarly, injection of an anti-chemokine ligand 2 (CCL2) antibody induced CCL2 depletion. Flow cytometry and immunofluorescence were used to characterize the monocytes recruited to the thrombus. In vivo, absence of Gas6 was associated with a reduction of monocyte recruitment in both deep venous thrombosis models. Global monocyte depletion by clodronate leads to smaller thrombi in wild-type mice. Compared with wild type, the thrombi from Gas6-/- mice contain less inflammatory (CCR2hiCX3CR1lo) monocytes, consistent with a Gas6-dependent recruitment of this monocyte subset. Correspondingly, selective depletion of CCR2hiCX3CR1lo monocytes reduced the formation of venous thrombi in wild-type mice demonstrating a predominant role of the inflammatory monocytes in thrombosis. In vitro, the expression of both CCR2 and CCL2 were Gas6 dependent in monocytes and endothelial cells, respectively, impacting monocyte migration. Moreover, Gas6-dependent CCL2 expression and monocyte migration were mediated via JNK (c-Jun N-terminal kinase). This study demonstrates that Gas6 specifically promotes the recruitment of inflammatory CCR2hiCX3CR1lo monocytes through the regulation of both CCR2 and CCL2 during deep venous thrombosis.


Observation of heterogeneous gene products by FT-ICR MS.
The ability of FT-ICR MS to resolve isotopic variants of intact proteins for each of the charge states formed by electrospray ionization offers a sensitive, rapid method for detecting "low mass" heterogeneity, where this is defined as the presence of structural variants differing in mass by 2 Da or less. Such heterogeneity may reflect biological or chemical modifications of structure or may result from the coexpression of related proteins from a multi-gene family. In the analytical approach described here, comparisons are made between observed isotopic distributions and those expected for predicted protein sequences. Close agreement is demonstrated for a homogeneous model protein, and the utility of the method has been evaluated in the study of mouse major urinary proteins (MUPs), a group of closely related sequences. Divergence of the experimental isotopic distribution from distributions predicted for known MUP sequences can be explained, in quantitative terms, by the coexpression of closely related sequences. This approach provides a facile method for the assessment of protein homogeneity and for the detection of structural variants, without recourse to proteolytic digestion and analysis of the resulting products.


Complementary and alternative medicine use at a single pediatric inflammatory bowel disease center.
The aim of this study was to determine the use of complementary and alternative medicine (CAM) in an inflammatory bowel disease population at a single pediatric center. The secondary aims were to determine predictors of CAM use and assess parental attitude to CAM use. A survey was developed that was distributed electronically and given out in the clinic. Two hundred thirty-five surveys were analyzed. Thirty-six percent of respondents reported that their children had used CAM, while 19.6% were current users. Sixty-three percent of respondents were "extremely" or "very supportive" of CAM and 57.6% would have been "extremely" or "very supportive" at the time of their children's diagnosis. The most commonly used CAM modalities were fish oil (48.8%), probiotics (22.5%), acupuncture/pressure (17.5%), aloe (16.3%), yoga/meditation (16.3%), chiropractic (12%), and herbal medicine (13.8%). Multivariate analysis revealed 2 independent factors predictive of subjects using CAM: use of biologics (odds ratio of 2.8; p = .008) and subjects' parent using CAM (odds ratio of 10.9; p ≤ .001). More than one third of children in this study and their parents have used CAM. Families are supportive of CAM both at the time of diagnosis and as an ongoing component of their child's treatment even if they were not past or current users of CAM. Predictors of CAM use were treatment with a biologic and having a parent who used CAM.


The relationship of the bayes risk to certain separability measures in normal classification.
For the problem of classifying an element (e.g., an unknown pattern) into one of two given categories where the associated observables are distributed according to one of two known multivariate normal populations having a common covariance matrix, it is shown that the minimum Bayes risk is a strict monotonic function of certain separability or statistical distance measures regardless of the a priori probabilities and the assigned loss function. However, for the associated conditional expected losses, strict monotonicity holds, if and only if a certain condition dependent on these probabilities and the given loss function is satisfied. These results remain valid for classification problems in which the observable can be transformed by a one-to-one differentiable mapping to normality.


Endogenous gastric-resident mesenchymal stem cells contribute to formation of cancer stroma and progression of gastric cancer.
Carcinoma-associated fibroblasts (CAFs) contribute to carcinogenesis and cancer progression, although their origin and role remain unclear. We recently identified and investigated the in situ identity and implications of gastric submucosa-resident mesenchymal stem cells (GS-MSCs) in the progression of gastric carcinogenesis. We isolated GS-MSCs from gastric submucosa using hydrogel-supported organ culture and defined their identity. Isolated cells were assessed in vitro by immunophenotype and mesengenic multipotency. Reciprocal interactions between GS-MSCs and gastric cancer cells were evaluated. To determine the role of GS-MSCs, xenografts were constructed of gastric cancer cells admixed with or without GS-MSCs. Isolated cells fulfilled MSCs requirements in regard to plastic adherence, stromal cell immunophenotype, and multipotency. We demonstrated a paracrine loop that gastric cancer cells enhanced the migration, proliferation, and differentiation of GS-MSCs; additionally, GS-MSCs promoted the proliferation of gastric cancer cell in vitro. Xenograft experiments showed that GS-MSCs significantly promoted cancer growth and angiogenesis. GS-MSCs that integrated into gastric cancer became not only CAFs but also rarely endothelial cells which contributed to the formation of cellular and vascular cancer stroma. Endogenous GS-MSCs play an important role in gastric cancer progression.


Incidence, Microbiological Profile, and Impact of Preventive Measures on Central Line-associated Bloodstream Infection in Liver Care Intensive Care Unit.
Central line-associated bloodstream infection (CLABSI) is among one of the preventable healthcare-associated infections (HAIs). The data for the CLABSI rate in liver care intensive care unit (LCICU) patients are scarce, so the present study was conducted to ascertain the CLABSI rate, the microbiological profile, and the impact of preventive measures for reduction of infection. This is a prospective observational study done on LCICU patients during the period of January 2017-December 2018. We followed up patients on the central venous catheter for the development of CLABSI as a part of routine surveillance of HAIs. The impact of introduction and implementation of the CLABSI bundle to reduce the CLABSI rate was analyzed and the microbiological profile of infection was determined. During the study period, the total number of patients admitted in LCICU were 1,336 (648 in 2017 and 688 in 2018) and a total of 995 central lines were inserted for various indications. A total of 57 patients were meeting the CLABSI criteria among 7,324 central line catheter days of surveillance. In year 2017, rate of CLABSI was 11.78/1,000 catheter days and after implementation of the bundle in 2018 the rate reduced to 3.99/1,000 catheter days. Gram-negative organisms (86%) predominated with Pseudomonas aeruginosa being the most common pathogen (19.3%). Out of 49 isolates of gram-negative bacilli (GNB), 40 (81.6%) were multidrug resistant (MDR) and 9 (18.4%) were pan-drug resistant. We found significant reduction in the CLABSI rate after implementation of the bundle of care. Gram-negative bacilli were the most common pathogen in our study and antimicrobial resistance was very high, which suggest hospital environment as a source of infection. Knowledge of the microbiological profile and the preventive strategy of CLABSI is essential for prevention and timely initiation of the most appropriate anti-infective therapy, if it happens. Khodare A, Kale P, Pindi G, Joy L, Khillan V. Incidence, Microbiological Profile, and Impact of Preventive Measures on Central Line-associated Bloodstream Infection in Liver Care Intensive Care Unit. Indian J Crit Care Med 2020;24(1):17-22.


Ultrafine particle levels measured on board short-haul commercial passenger jet aircraft.
Airline crew members report adverse health effects during and after inhalation exposure to engine oil fumes sourced to the air supply system onboard commercial and military aircraft. Most investigations into the causal factors of their reported symptoms focus on specific chemical contaminants in the fumes. The adverse health effects reported in aircrew exposed to the aircraft air supply, bled unfiltered off the engine or Auxiliary Power Unit (APU) may be related to particulate exposures, which are widely known to effect health. While oil contaminates the aircraft air supply, some suggest that this will only occur when there is a bearing seal failure, others document that there is low level oil contamination of the air supply during normal engine operation. This brief pilot study explores whether particulate exposure may be associated with the normal engine/APU and air supply operation and to therefore increase the understanding that UFP exposures may have on crew and passengers. An ultrafine particle counter was utilised by an experienced airline captain in the passenger cabin of four short-haul commercial passenger aircraft. All flights were under 90 min on aircraft from two different carriers ranging from 7 months to 14 years old. UFP concentrations showed maximum concentrations ranging from 31,300 to 97,800 particles/cm3 when APU was selected on as a source of air on the ground and with engine bleed air and the air conditioning packs selected on during the climb. In 2 of the 4 flights the peaks were associated with an engine oil smell. Increases in UFP particle concentrations occurred with changes in engine/APU power and air supply configuration changes. This study identified increases in UFP concentrations associated with engine and APU power changes and changes in air supply configuration. These results correlated with times when engine and APU oil seals are known to be less effective, enabling oil leakage to occur. The concentrations reached in the passenger cabins exceeded those taken in other ground-based environments. UFP exposures in aircraft cabins during normal flight indicates there will be health consequences for long serving aircrew and some passengers.


Karyotype analysis of the euploid cell population of a mouse embryonic stem cell line revealed a high incidence of chromosome abnormalities that varied during culture.
It is common knowledge that mouse embryonic stem cell (mESC) lines accumulate chromosomal changes during culture. Despite the wide use of mESCs as a model of early mammalian development and cell differentiation, there is a lack of systematic studies aimed at characterizing their karyological changes during culture. We cultured an mESC line, derived in our laboratory, for a period of 3 months investigating its chromosome complement at different times. About 60% of the metaphases analysed were euploid throughout the culture period but, from passage 13, only 50% of the euploid metaphases had a proper chromosome complement. The remaining 50% showed chromosome abnormalities, mainly gain or loss of entire chromosomes, both within the same passage and among different passages analysed. The very heterogeneous spectrum of abnormalities indicates a high frequency of chromosome mutations that arise continuously during culture. The heterogeneity of the aberrant chromosome constitution of 2n = 40 metaphases, observed at different passages of culture, might be due either to their elimination or to a shift towards the hypoeu- or hypereuploid population of those metaphases that accumulate further chromosome abnormalities. The stability of the frequency of eu-, hypoeu- and hypereuploid populations during culture might, however, be due to the elimination of those cells that carry a high mutational burden. Based on our results, we suggest that karyotype analysis of the euploid cell population of mESC lines is necessary when such lines are used in the production of chimeric mice, for their contribution to the germ line, or when they are differentiated into specific cell types.


Structural and evolutionary relationships of beta-lactamase transposons from Staphylococcus aureus.
A comparison of the beta-lactamase elements detected on three classes of large plasmids together with the chromosomes of penicillin-resistant Staphylococcus aureus revealed substantial physical and genetic relatedness. In most cases, beta-lactamase production could be associated with the presence of a DNA segment of approximately 6.7 kb. Analysis showed that the plasmid-borne determinants constitute nearly identical transposons or transposon-like elements. An element indistinguishable from one of these, Tn4002, which is carried by the pSK1 family of plasmids in clinical isolates from Australian hospitals, was also identified on the staphylococcal chromosome and is implicated in an evolutionary cycle of transposition between chromosomal and extrachromosomal sites in Australian strains of multiresistant S. aureus.


Gene expression in plant lipid metabolism in Arabidopsis seedlings.
Events in plant lipid metabolism are important during seedling establishment. As it has not been experimentally verified whether lipid metabolism in 2- and 5-day-old Arabidopsis thaliana seedlings is diurnally-controlled, quantitative real-time PCR analysis was used to investigate the expression of target genes in acyl-lipid transfer, β-oxidation and triacylglycerol (TAG) synthesis and hydrolysis in wild-type Arabidopsis WS and Col-0. In both WS and Col-0, ACYL-COA-BINDING PROTEIN3 (ACBP3), DIACYLGLYCEROL ACYLTRANSFERASE1 (DGAT1) and DGAT3 showed diurnal control in 2- and 5-day-old seedlings. Also, COMATOSE (CTS) was diurnally regulated in 2-day-old seedlings and LONG-CHAIN ACYL-COA SYNTHETASE6 (LACS6) in 5-day-old seedlings in both WS and Col-0. Subsequently, the effect of CIRCADIAN CLOCK ASSOCIATED1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY) from the core clock system was examined using the cca1lhy mutant and CCA1-overexpressing (CCA1-OX) lines versus wild-type WS and Col-0, respectively. Results revealed differential gene expression in lipid metabolism between 2- and 5-day-old mutant and wild-type WS seedlings, as well as between CCA1-OX and wild-type Col-0. Of the ACBPs, ACBP3 displayed the most significant changes between cca1lhy and WS and between CCA1-OX and Col-0, consistent with previous reports that ACBP3 is greatly affected by light/dark cycling. Evidence of oil body retention in 4- and 5-day-old seedlings of the cca1lhy mutant in comparison to WS indicated the effect of cca1lhy on storage lipid reserve mobilization. Lipid profiling revealed differences in primary lipid metabolism, namely in TAG, fatty acid methyl ester and acyl-CoA contents amongst cca1lhy, CCA1-OX, and wild-type seedlings. Taken together, this study demonstrates that lipid metabolism is subject to diurnal regulation in the early stages of seedling development in Arabidopsis.


An overview of new and emerging antibody therapies for moderate-severe atopic dermatitis in adults.
A comprehensive review of the English-language medical literature was performed searching for ongoing and closed clinical trials concerning new and emerging monoclonal antibody therapies for moderate-to-severe atopic dermatitis in adults. Atopic dermatitis is a chronic inflammatory cutaneous disease with a complex pathogenesis. In the last years, numerous advances in understanding the atopic dermatitis pathogenesis allowed to obtain several therapeutic options, such as numerous monoclonal antibodies. Some monoclonal antibodies, such as dupilumab (anti-IL-4 Rα) and tralokinumab (anti-IL13) are already approved for the treatment of moderate-to-severe atopic dermatitis, and numerous articles in the literature have demonstrated their efficacy and safety. As there are numerous drugs under investigation, this review focuses on emerging monoclonal antibody therapies. There are numerous monoclonal antibodies under investigation that may be approved in the near future for the treatment of atopic dermatitis. Data from phase 2b and phase III clinical trials in moderate-to-severe atopic dermatitis in adults indicate that these drugs have a promising efficacy and safety profile. Monoclonal antibodies currently under investigation will be available in the coming years to enrich the therapeutic choice of new alternatives that are valid both in terms of efficacy and safety.


[Cycloferon biological activity characteristics].
Study the effect of cycloferon in experimental and clinical conditions on persistence properties of aurococci as well as features of their morpho-functional reaction by atomic force microscopy. The study was carried out in 12 Staphylococcus aureus clones isolated from mucous membrane of nose anterior part of a resident carrier. The effect of cycloferon in vivo was evaluated in 26 resident staphylococci carriers under the control of anti-carnosine activity of staphylococci. Anti-carnosine activity was determined by O.V. Bukharin et al. (1999), biofilm formation -by G.A. O'Toole et al. (2000). Staphylococci treated with cycloferon were studied by atomic force microscopy in contact mode using scanning probe SMM-2000 microscope. The decrease of persistence properties of staphylococci under the effect of cycloferon in vitro and in vivo may be examined as one of the mechanisms of biological activity of the preparation. A significant increase of S. aureus surface roughness and changes in their morphology under the effect of cycloferon allow stating the disorder of barrier functions in the aurococci cell wall. The data obtained expand the understanding of cycloferon biological activity mechanisms.


Nitrate- and molybdenum-independent signal transduction mutations in narX that alter regulation of anaerobic respiratory genes in Escherichia coli.
Escherichia coli can respire anaerobically by reducing nitrate, trimethylamine-N-oxide, dimethyl sulfoxide, or fumarate. When nitrate is present, expression of the genes for fumarate (frdABCD), trimethylamine-N-oxide, and dimethyl sulfoxide (dmsABC) is repressed while expression of the nitrate reductase (narGHJI) gene is induced. This regulation requires molybdate and is mediated by the narX and narL gene products, which together form a two-component regulatory system. We provide evidence that NarX is a nitrate and molybdenum sensor which activates NarL when nitrate is available to cells. Mutants generated by hydroxylamine mutagenesis were repressed for frdA-lacZ expression even when cells were grown in the absence of nitrate. The mutations responsible for three of these nitrate independence (NarX*) phenotypes were localized to narX and further characterized in vivo for their ability to repress frdA-lacZ expression. Two of the mutants (the narX64 and narX71 mutants) had a greatly reduced requirement for molybdenum to function but still responded to nitrate. In contrast, a third mutant (the narX32 mutant) required molybdenum but did not exhibit full repression of frdA-lacZ expression even when nitrate was present. These narX* alleles also caused the induction of nitrate reductase gene expression and the repression of a dmsA-lacZ fusion in the absence of nitrate. Each narX* mutation was determined to lie in an 11-amino-acid region of the NarX polypeptide that follows a proposed transmembrane domain. We suggest that the conformation of the narX* gene products is altered such that even in the absence of nitrate each of these gene products more closely resembles the wild-type NarX protein when nitrate is present. These data establish a clear role for the narX gene product in gene regulation and strongly suggest its role in sensing nitrate and molybdenum.


Mandated reporters' experiences with reporting child maltreatment: a meta-synthesis of qualitative studies.
To systematically search for research about the effectiveness of mandatory reporting of child maltreatment and to synthesise qualitative research that explores mandated reporters' (MRs) experiences with reporting. As no studies assessing the effectiveness of mandatory reporting were retrieved from our systematic search, we conducted a meta-synthesis of retrieved qualitative research. Searches in Medline (Ovid), Embase, PsycINFO, Cumulative Index to Nursing and Allied Health Literature, Sociological Abstracts, Education Resources Information Center, Criminal Justice Abstracts and Cochrane Library yielded over 6000 citations, which were deduplicated and then screened by two independent reviewers. English-language, primary qualitative studies that investigated MRs' experiences with reporting of child maltreatment were included. Critical appraisal involved a modified checklist from the Critical Appraisal Skills Programme and qualitative meta-synthesis was used to combine results from the primary studies. All healthcare and social-service settings implicated by mandatory reporting laws were included. Included studies crossed nine high-income countries (USA, Australia, Sweden, Taiwan, Canada, Norway, Finland, Israel and Cyprus) and three middle-income countries (South Africa, Brazil and El Salvador). The studies represent the views of 1088 MRs. Factors that influence MRs' decision to report and MRs' views towards and experiences with mandatory reporting of child maltreatment. Forty-four articles reporting 42 studies were included. Findings indicate that MRs struggle to identify and respond to less overt forms of child maltreatment. While some articles (14%) described positive experiences MRs had with the reporting process, negative experiences were reported in 73% of articles and included accounts of harm to therapeutic relationships and child death following removal from their family of origin. The findings of this meta-synthesis suggest that there are many potentially harmful experiences associated with mandatory reporting and that research on the effectiveness of this process is urgently needed.


Understanding the Reasons for Suicide Among Older Adults in Rural China Using In-Depth Interviews.
Background: Suicide is a complex public health problem. Suicide rates are higher in rural areas than in urban areas and among older adults than any other age group in China. Aims: This study aimed to acquire a comprehensive understanding of rural older adults' reasons for suicide through qualitative exploration and to provide additional insights for suicide prevention work. Method: In-depth interviews were conducted with people familiar with older adults who had died by suicide ("informants"). The data were analyzed using a thematic approach. Results: Data were available for 242 people who died by suicide. Informants identified two or more reasons for suicide among most people who had died. Physical illness, psychological distress, and interpersonal conflicts were the most common reasons. The rate of mental disorders as diagnosed by psychiatrists was higher than that reported by the informants. Limitations: Information bias is inevitable because of the use of proxy informants. Conclusion: The reasons for suicide are complex and diverse, and research on suicide should focus more on the mechanisms of interaction between reasons. Greater attention should be paid to physical and mental health as well as to improving interpersonal and problem-solving skills for older adults in China.


Rate of muscle activation in power- and endurance-trained boys.
Previous studies in adults have demonstrated power athletes as having greater muscle force and muscle activation than nonathletes. Findings on endurance athletes are scarce and inconsistent. No comparable data on child athletes exist. This study compared peak torque (Tq), peak rate of torque development (RTD), and rate of muscle activation (EMG rise, Q30), in isometric knee extension (KE) and flexion (KF), in pre- and early-pubertal power- and endurance-trained boys vs minimally active nonathletes. Nine gymnasts, 12 swimmers, and 18 nonathletes (7-12 y), performed fast, maximal isometric KE and KF. Values for Tq, RTD, electromechanical delay (EMD), and Q30 were calculated from averaged torque and surface EMG traces. No group differences were observed in Tq, normalized for muscle cross-sectional area. The Tq-normalized KE RTD was highest in power athletes (6.2 ± 1.9, 4.7 ± 1.2, 5.0 ± 1.5 N·m·s-1, for power, endurance, and nonathletes, respectively), whereas no group differences were observed for KF. The KE Q30 was significantly greater in power athletes, both in absolute terms and relative to peak EMG amplitude (9.8 ± 7.0, 5.9 ± 4.2, 4.4 ± 2.2 mV·ms and 1.7 ± 0.8, 1.1 ± 0.6, 0.9 ± 0.5 (mV·ms)/(mV) for power, endurance, and nonathletes, respectively), with no group differences in KF. The KE EMD tended to be shorter (P = .07) in power athletes during KE (71.0 ± 24.1, 87.8 ± 18.0, 88.4 ± 27.8 ms, for power, endurance, and nonathletes), with no group differences in KF. Pre- and early-pubertal power athletes have enhanced rate of muscle activation in specifically trained muscles compared with controls or endurance athletes, suggesting that specific training can result in muscle activation-pattern changes before the onset of puberty.


Parental Corporal Punishment and Peer Victimization in Middle Childhood: A Sex-Moderated Mediation Model of Aggression.
There is a peak in peer victimization during middle childhood, with multiple negative consequences. Parental use of corporal punishment and child aggression are the most widely studied predictors of this phenomenon. The aim of the present study was to analyze whether parental use of corporal punishment affects peer victimization through child aggression. This mediation model was explored for both mothers and fathers and for both physical and relational forms of aggression and peer victimization. Furthermore, we also analyzed whether the mediation models were moderated by the sex of the child. Participants were 234 third graders (46% girls). Child aggression and victimization were measured by peers using the Mini Direct Indirect Aggression Inventory. Independent measures of mother's and father's use of corporal punishment were obtained from a PCA of items from the Parental Styles and Dimensions Questionnaire (PSDQ). Conditional process modeling was carried out using a macro for SPSS developed by Hayes (2013). Results indicated that aggression mediated the relation of parental corporal punishment to peer victimization. Some interesting moderating effects of sex in this mediation model were found; specifically, physical, and relational aggression mediated the relation of maternal corporal punishment to peer victimization only in boys. Few studies to date have addressed the connection between aggressive behavior and peer victimization as outcomes of corporal punishment, taking into consideration the role of parent's and child's sex, and both physical and relational forms of aggression and victimization during childhood.


Isoflurane depolarizes bronchopulmonary C neurons by inhibiting transient A-type and delayed rectifier potassium channels.
Inhalation of isoflurane (ISO), a widely used volatile anesthetic, can produce clinical tachypnea. In dogs, this response is reportedly mediated by bronchopulmonary C-fibers (PCFs), but the relevant mechanisms remain unclear. Activation of transient A-type potassium current (IA) channels and delayed rectifier potassium current (IK) channels hyperpolarizes neurons, and inhibition of both channels by ISO increases neural firing. Due to the presence of these channels in the cell bodies of rat PCFs, we determined whether ISO could stimulate PCFs to produce tachypnea in anesthetized rats, and, if so, whether this response resulted from ISO-induced depolarization of the pulmonary C neurons via the inhibition of IA and IK. We recorded ventilatory responses to 5% ISO exposure in anesthetized rats before and after blocking PCF conduction and the responses of pulmonary C neurons (extracellularly recorded) to ISO exposure. ISO-induced (1mM) changes in pulmonary C neuron membrane potential and IA/IK were tested using the perforated patch clamp technique. We found that: (1) ISO inhalation evoked a brief tachypnea (∼7s) and that this response disappeared after blocking PCF conduction; (2) the ISO significantly elevated (by 138%) the firing rate of most pulmonary C neurons (17 out of 21) in the nodose ganglion; and (3) ISO perfusion depolarized the pulmonary C neurons in the vitro and inhibited both IA and IK, and this evoked-depolarization was largely diminished after blocking both IA and IK. Our results suggest that ISO is able to stimulate PCFs to elicit tachypnea in rats, at least partly, via inhibiting IA and IK, thereby depolarizing the pulmonary C neurons.


Listeria monocytogenes prosthetic valve endocarditis.
Listeria monocytogenes is a rare cause of prosthetic valve endocarditis and in all of the four reported cases has occurred as a late complication. Bacteriologic cure of the infection has been obtained in all reported patients, using a standard regimen of either penicillin or penicillin and an aminoglycoside. The two deaths were associated with Starr-Edwards prostheses in the aortic position and have been attributed to embolic phenomena occurring after bacteriologic sterilization of the infected valve.


Pseudomonas aeruginosa displays a dormancy phenotype during long-term survival in water.
Pseudomonas aeruginosa is capable of long-term survival in water, which may serve as a reservoir for infection. Although viable cell counts of PAO1 incubated in water remain stable throughout 8 weeks, LIVE/DEAD staining indicated a high proportion of cells stained with propidium iodide (PI). The proportion of PI-stained cells increased by 4 weeks, then decreased again by 8 weeks, suggesting an adaptive response. This was also evident in an observed shift in cell morphology from a rod to a coccoid shape after 8 weeks. Fluorescence-activated cell sorting (FACS) was used to recover PI-stained cells, which were plated and shown to be viable, indicating that PI-stained cells were membrane-compromised but still cultivable. PAO1 mid-log cells in water were labeled with the dsDNA-binding dye PicoGreen to monitor viability as well as DNA integrity, which demonstrated that the population remains viable and transitions towards increased dsDNA staining. Metabolic activity was found to decrease significantly in water by 4 weeks. The PAO1 outer membrane became less permeable and more resistant to polymyxin B damage in water, and the profile of total membrane lipids changed over time. Among the ~1400 transcriptional lux fusions, gene expression in water revealed that the majority of genes were repressed, but subsets of genes were induced at particular time points. In summary, these results indicate that P. aeruginosa is dormant in water and this adaptation involves a complex pattern of gene regulation and changes to the cell to promote long-term survival and antibiotic tolerance. The approach of P. aeruginosa incubated in water may be useful to study antibiotic tolerance and the mechanisms of dormancy and survival in nutrient limiting conditions.


Reliability of breeding values for feed intake and feed efficiency traits in dairy cattle: When dry matter intake recordings are sparse under different scenarios.
Currently, routine recordings of dry matter intake (DMI) in commercial herds are practically nonexistent. Recording DMI from commercial herds is a prerequisite for the inclusion of feed efficiency (FE) traits in dairy cattle breeding goals. To develop future on-farm phenotyping strategies, recording strategies that are low cost and less demanding logistically and that give relatively accurate estimates of the animal's genetic merit are therefore needed. The objectives of this study were (1) to estimate genetic parameters for daily DMI and FE traits and use the estimated parameters to simulate daily DMI phenotypes under different DMI recording scenarios (SCN) and (2) to use the simulated data to estimate for different scenarios the associated reliability of estimated breeding value and accuracies of genomic prediction for varying sizes of reference populations. Five on-farm daily DMI recording scenarios were simulated: once weekly (SCN1), once monthly (SCN2), every 2 mo (SCN3), every 3 mo (SCN4), and every 4 mo (SCN5). To estimate reliability of estimated breeding values, DMI and FE observations and true breeding values were simulated based on variance components estimated for daily observations of Nordic Red cows. To emulate realistic on-farm recording, 5 data set replicates, each with 36,037 DMI and FE records, were simulated for real pedigree and data structure of 789 Holstein cows. Observations for the 5 DMI recording scenarios were generated by discarding data in a step-wise manner from the full simulated data per the scenario's definitions. For each of these scenarios, reliabilities were calculated as correlation between the true and estimated breeding values. Variance components and genetic parameters were estimated for daily DMI, residual feed intake (RFI), and energy conversion efficiency (ECE) fitting the random regression model. Data for variance components were from 227 primiparous Nordic Red dairy cows covering 8 to 280 d in milk. Lactation-wise heritability for DMI, RFI, and ECE was 0.33, 0.12, and 0.32, respectively, and daily heritability estimates during lactation ranged from 0.18 to 0.45, 0.08 to 0.32, and 0.08 to 0.45 for DMI, RFI, and ECE, respectively. Genetic correlations for DMI between different stages of lactation ranged from -0.50 to 0.99. The comparison of different on-farm DMI recording scenarios indicated that adopting a less-frequent recording scenario (SCN3) gave a similar level of accuracy as SCN1 when 17 more daughters are recorded per sire over the 46 needed for SCN1. Such a strategy is less demanding logistically and is low cost because fewer observations need to be collected per animal. The accuracy of genomic predictions associated with the 5 recording scenarios indicated that setting up a relatively larger reference population and adopting a less-frequent DMI sampling scenario (e.g., SCN3) is promising. When the same reference population size was considered, the genomic prediction accuracy of SCN3 was only 5.0 to 7.0 percentage points lower than that for the most expensive DMI recording strategy (SCN1). We concluded that DMI recording strategies that are sparse in terms of records per cow but with slightly more cows recorded per sire are advantageous both in genomic selection and in traditional progeny testing schemes when accuracy, logistics, and cost implications are considered.


Risk Factors for Cerebrospinal Fluid Leakage After Extradural Spine Surgery: A Meta-Analysis and Systematic Review.
Cerebrospinal fluid (CSF) leakage is 1 of the common complications of spine surgery and is largely caused by intraoperative or postoperative dural tears. Associations of different factors with postoperative CSF leakage have not been consistent. In this study we aimed to identify demographic, disease-related, and surgical risk factors for CSF leakage after extradural spine surgery in a systematic review and meta-anlysis. The PubMed, EMBASE, Web of Science, Cochrane Library, Chinese National Knowledge Infrastructure, Chinese Wanfang data, Chinese Weipu Database, and SinoMed databases were searched from inception until October 24, 2022. Fixed-effects or random-effects models were used to calculate odds ratios and 95% confidence intervals. The quality of observational studies was evaluated using the Newcastle-Ottawa scale instrument. A total of 15 observational studies with 1,719,923 participants were included in this systematic review. All studies had a Newcastle-Ottawa scale score greater than or equal to 6. Age older than 70 years, smoking, ossification of the posterior longitudinal ligament, adhesion of spinal dura, spinal canal stenosis, cervical fracture, spondylolisthesis, revision surgery, and multiple surgical segments were all related to CSF leakage in the pooled analysis. Obesity and disease duration>1 year were not associated with the leakage of CSF. This study will provide a reference for the identification of patients at high risk of developing CSF leakage, which suggests clinicians to strengthen the observation of drainage fluid in high-risk groups.


First-in-Man Study Evaluating the Emblok Embolic Protection System During Transcatheter Aortic Valve Replacement.
This study sought to evaluate the feasibility of complete cerebral protection during transcatheter aortic valve replacement (TAVR) with a novel embolic protection device. Evidences and data about new cerebral embolic protection devices are lacking and scarce. A prospective, nonrandomized, multicenter, first-in-man pilot study designed to evaluate the efficacy and safety of cerebral embolic protection utilizing the Emblok embolic protection system (Innovative Cardiovascular Solutions, Grand Rapids, Michigan) during TAVR. The Emblok is a transfemoral aortic filter that provide full coverage of the epiaortic vessels. Brain diffusion-weighted magnetic resonance imaging (DW-MRI) was performed at baseline and 2 to 5 days after TAVR. Primary endpoints were technical success and immediate cerebral embolic burden after TAVR, defined as number and volume of new brain lesions detected with DW-MRI at days 2 to 5 post-TAVR compared with baseline. A total of 20 subjects were enrolled. The Emblok system was successfully positioned in all the cases. At 30-day follow-up, no major adverse cardiovascular and cerebrovascular events occurred. Nineteen (95%) patients had new ischemic defects at post-procedural DW-MRI. The median number of new lesions per patient was 10.00 (interquartile range [IQR]: 4.75 to 15.25). The total new lesion volume was 199.9 mm3 (IQR: 83.9 to 447.5 mm3) and the mean lesion volume per lesion was 42.5 mm3 (IQR: 21.5 to 75.6 mm3). Histopathologic analysis showed evidence of significant debris in 18 (90%) filters. The Emblok embolic protection system appears to be feasible and safe during TAVR. The device was successfully placed and retrieved in all cases and no neurological events were observed. Cerebral total new lesion volume was similar to other trials on cerebral protection during TAVR, thus warranting a larger study. (European Study Evaluating the Emblok Embolic Protection System During TAVR; NCT03130491).


Genome sequencing of Giardia lamblia genotypes A2 and B isolates (DH and GS) and comparative analysis with the genomes of genotypes A1 and E (WB and Pig).
Giardia lamblia (syn G. intestinalis, G. duodenalis) is the most common pathogenic intestinal parasite of humans worldwide and is a frequent cause of endemic and epidemic diarrhea. G. lamblia is divided into eight genotypes (A-H) which infect a wide range of mammals and humans, but human infections are caused by Genotypes A and B. To unambiguously determine the relationship among genotypes, we sequenced GS and DH (Genotypes B and A2) to high depth coverage and compared the assemblies with the nearly completed WB genome and draft sequencing surveys of Genotypes E (P15; pig isolate) and B (GS; human isolate). Our results identified DH as the smallest Giardia genome sequenced to date, while GS is the largest. Our open reading frame analyses and phylogenetic analyses showed that GS was more distant from the other three genomes than any of the other three were from each other. Whole-genome comparisons of DH_A2 and GS_B with the optically mapped WB_A1 demonstrated substantial synteny across all five chromosomes but also included a number of rearrangements, inversions, and chromosomal translocations that were more common toward the chromosome ends. However, the WB_A1/GS_B alignment demonstrated only about 70% sequence identity across the syntenic regions. Our findings add to information presented in previous reports suggesting that GS is a different species of Giardia as supported by the degree of genomic diversity, coding capacity, heterozygosity, phylogenetic distance, and known biological differences from WB_A1 and other G. lamblia genotypes.


Environmental unknowns.
Among the environmental problems ahead, the most important ones could be those that are still unknown to us. This conceptual article explores this prospect on the grounds that it is important not only to supply answers to recognized questions but to raise appropriate new questions.


Neuropeptidergic integration of behavior in Trichoplax adhaerens, an animal without synapses.
Trichoplax adhaerens is a flat, millimeter-sized marine animal that adheres to surfaces and grazes on algae. Trichoplax displays a repertoire of different feeding behaviors despite the apparent absence of a true nervous system with electrical or chemical synapses. It glides along surfaces to find food, propelled by beating cilia on cells at its ventral surface, and pauses during feeding by arresting ciliary beating. We found that when endomorphin-like peptides are applied to an animal, ciliary beating is arrested, mimicking natural feeding pauses. Antibodies against these neuropeptides label cells that express the neurosecretory proteins and voltage-gated calcium channels implicated in regulated secretion. These cells are embedded in the ventral epithelium, where they comprise only 4% of the total, and are concentrated around the edge of the animal. Each bears a cilium likely to be chemosensory and used to detect algae. Trichoplax pausing during feeding or spontaneously in the absence of food often induce their neighbors to pause as well, even neighbors not in direct contact. Pausing behavior propagates from animal to animal across distances much greater than the signal that diffuses from just one animal, so we presume that the peptides secreted from one animal elicit secretion from nearby animals. Signal amplification by peptide-induced peptide secretion explains how a small number of sensory secretory cells lacking processes and synapses can evoke a wave of peptide secretion across the entire animal to globally arrest ciliary beating and allow pausing during feeding.


The Effect of Gelatin Molecular Weight on Tendon Lubrication Utilizing an Extrasynovialized Turkey Flexor Tendon Model.
Flexor tendon injuries are common hand injuries among the military population often resulting in functional impairment. Flexor tendon gliding friction has been linked to adhesion formation, especially with the use of extrasynovial grafts. Carbodiimide-derivatized hyaluronic acid with gelatin (cd-HA-gelatin) can reduce gliding friction of the tendon graft; however, the effects of gelatin molecular weight (MW) have not been studied. The turkey model has been shown to better match humans, but extrasynovial tendons are unavailable. The purpose of this study was to (1) manually roughen turkey flexor digitorum profundus (FDP) tendons to simulate extrasynovial tendons and (2) investigate the effects of gelatin MW on tendon friction. The third digit of (n = 48) turkeys were dissected with the proximal pulley, and FDP tendon and the flexor superficialis tendon were preserved. Digits were randomly assigned into four groups: one saline control and three cd-HA-gelatin-treated groups of varying gelatin MW. Flexor digitorum profundus tendon friction was measured at its original condition, serving as a baseline. Tendons were roughened using a custom rig, and tendon friction was measured again. All four groups received treatment and gliding friction was measured every 100 cycles to a total of 1,000 cycles. Tendon friction significantly increased (P < 0.05) after roughening. Friction in the saline control group increased steadily over repeated cycles, whereas friction of all gelatin-treated tendons decreased significantly compared with the saline control group (P < 0.05), maintaining low friction to 1,000 cycles representing human tendons. There was no significant difference found between gelatin-treated groups. We have developed a method to roughen synovial FDP tendons to create extrasynovial-like tendons for lubrication material evaluations. Cd-HA-gelatin effectively reduces tendon friction in this model. Our data suggest medium or low MW gelatin may provide a better reduction in friction compared with high MW gelatin.


The role of associative and non-associative learning in the training of horses and implications for the welfare (a review).
Horses were domesticated 6000 years ago and since then different types of approaches have been developed to enhance the horse's wellbeing and the human-horse relationship. Even though horse training is an increasingly important research area and many articles have been published on the subject, equitation is still the sport with the highest rate of human injuries, and a significant percentage of horses are sold or slaughtered due to behavioral problems. One explanation for this data is that the human-horse relationship is complex and the communication between humans and horses has not yet been accurately developed. Thus, this review addresses correct horse training based on scientific knowledge in animal learning and psychology. Specifically, it starts from the basic communication between humans and horses and then focuses on associative and non-associative learning, with many practical outcomes in horse management from the ground and under saddle. Finally, it highlights the common mistakes in the use of negative reinforcement, as well as all the implications that improper training could have on horse welfare. Increased levels of competence in horse training could be useful for equine technicians, owners, breeders, veterinarians, and scientists, in order to safeguard horse welfare, and also to reduce the number of human injuries and economic loss for civil society and the public health system.


[Stimulation of degenerative changes in the intervertebral disc through axial compression. Radiologic, histologic and biomechanical research in an animal model].
Degeneration of the intervertebral disc is a common disease in the adults, especially at advanced age. A causal therapy is not known, but the progress in new therapeutic strategies, for example in tissue engineering, shows new possibilities. The goal of our study was to develop a new animal model that stimulates a load induced degeneration of the disc. We used the New Zealand rabbit, because morphology is similar to the human intervertebral disc. The degeneration was induced by axial compression of the disc L4 - L5 with an external fixateur. After different loading intervals, the animals were sacrified and the discs examined by radiology, histology, apoptosis and biomechanical testing. Radiography showed a significant decrease of the disc thickness in all loaded groups. Morphologically the intervertebral discs of loaded rabbits showed degenerative changes which were comparable to those in humans. A significantly increased number of dead cells in the annulus occurred after 14 and 28 days loading compared to the controls. The bending stress measured as the load to failure was not significantly different between the unloaded discs and the 28 days loaded discs. The results show that our animal modell can create degeneration. Four weeks compression leads to significant degeneration. Degeneration of the discs persisted in animals that were allowed a recovery time of 28 days after 28 days of loading.


A rare but serious manifestation of Behçet's disease: intracardiac thrombus in 22 patients.
Behçet's disease (BD) is a chronic, multisystemic disorder characterised by recurrent oral aphtous ulcers, genital ulcers and ocular inflammation. Vasculitis and thrombotic events are the most important causes of mortality. Vena cava thrombosis, pulmonary artery aneurysms, Budd-Chiari syndrome, peripheral artery aneurysms, dural sinus thrombosis and abdominal aorta aneurysms are the other less common vascular manifestations of BD. Cardiac involvement in BD is a rare and life-threatening complication. The aim of this study was to assess the clinical characteristics and outcome of patients with BD who have intracardiac thrombus. The hospital files of BD patients followed by rheumatology clinics of four medical centres (Ankara University, Ege University, Hacettepe University and Eskişehir Osmangazi University Hospitals) were retrospectively evaluated. Data included patients' demographic and clinical features, laboratory findings and outcome. All patients fulfilled three or more of the International Study Group Criteria for BD. Twenty-two patients with intracardiac thrombus were evaluated. The mean age of patients with intracardiac thrombosis was 29.1 yrs (22-44) and there was a male predominance with a ratio of 20:2. Cardiac involvement was the first clinical manifestation of BD in 9 of the 22 patients. Initial symptoms of the patients were fever (n=18, 81%), dyspnea (n=9, 40%) chest pain (n=9, 40%) and haemoptysis (n=7, 31.8%). Sixteen patients (72%) had pulmonary arteritis and 10 (45%) patients had venous system lesions included deep vein, inferior vena cava and hepatic vein. Intra-cardiac thrombus were found only in the right cavities in 17 patients (77%), only in the left cavities in 2 patients (9%), and in both left and right cavities of the heart in 3 patients (13.6%). Once the cardiac lesion was diagnosed as a complication of BD, high dose (1mg/kg/d) prednisone (n=22, 100%) plus cyclophosphamide (n=18, 81%) or azathioprine (n=3, 13.6%) and warfarin (n=8, 36.3% (after the elimination of pulmonary aneurysm) therapy for anticoagulation was initiated. Four patients (18%) had high dose prednisone plus cyclophosphamide plus interferon-α (IFN-α) combination treatment and two patients (n=2, 9%) had high dose prednisone plus cyclophosphamide or plus azathioprine combination treatment. After treatment, the intra-cardiac thrombus disappeared in 13 cases and the size of the thrombus reduced in 7 cases. One patient died because of recurrent intra-cardiac thrombus and massive pulmonary arteritis in the emergency department. Intracardiac thrombus in BD is more common in young men. The right side of the heart is usually involved and cardiac involvement is often accompanied by pulmonary artery occlusion possibly due to pulmonary arteritis. Early and aggressive immunosuppressive and/or anticoagulation therapy are life-saving.


[Strategies for nurses to cope with the stress caused by working with mental patients].
This paper comprises a quantitative research to evaluate the coping mechanism in order to face stress in the nurse's work with the bearer of mental illness. It was conducted in seven psychiatric hospitals in the city of Fortaleza, Ceará, Brazil. The following instruments were used: socio-demographic data to describe the sample, an inventory in order to identify individual coping features. The sample comprised 42 participants, most of them female, corresponding to 92.9% of the total. Conclusion can be drawn that in order to deal with these stressing situations, the majority of the nurses used strategies focused on the problem, solving it when it arose or trying to review the situation with the possibility of an engaging attitude.


Characterization of fifty-one flavonoids in a Chinese herbal prescription Longdan Xiegan Decoction by high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry and photodiode array detection.
High-performance liquid chromatography coupled to electrospray ionization (ESI) tandem mass spectrometry and photodiode array detection (HPLC-DAD-ESI-MS(n)) was developed to identify and characterize the flavonoids in a Chinese formulated preparation, Longdan Xiegan Decoction (LXD). In total, fifty-one flavonoids (27 flavones, 10 flavanones, 7 chalcones, 5 flavonols and 2 isoflavones) were characterized. Eighteen compounds among them including a newly detected flavonoid, naringin, from the ingredient herbs, were unambiguously determined by comparing the retention times (t(R)), UV spectral data and mass fragmentation behaviors with those of the reference compounds. Another thirty-three compounds were tentatively identified by referencing to the reported data of their UV and MS spectra. The ESI-MS/MS fragmentation behavior of flavones (OMe-substituted, O-glycosides, C-glycosides), chalcones, flavonols and their appropriate characteristic pathways were proposed. In negative ion ESI-MS all the flavonoids yielded prominent [M--H](-) ions in the first order mass spectra. Fragmentation with a loss of mass of 15 Da (CH(3)), 18 Da (H(2)O), 28 Da (CO), 44 Da (CO(2)), 56 Da (2CO) and the residues of glucose and glucuronic acid observed in the MS/MS spectra were useful for aiding the structural identification of the flavonoids investigated.


Key Role of Capsular Polysaccharide in the Induction of Systemic Infection and Abortion by Hypervirulent Campylobacter jejuni.
Campylobacter jejuni is a zoonotic pathogen, and a hypervirulent clone, named clone SA, has recently emerged as the predominant cause of ovine abortion in the United States. To induce abortion, orally ingested Campylobacter must translocate across the intestinal epithelium, spread systemically in the circulation, and reach the fetoplacental tissue. Bacterial factors involved in these steps are not well understood. C. jejuni is known to produce capsular polysaccharide (CPS), but the specific role that CPS plays in systemic infection and particularly abortion in animals remains to be determined. In this study, we evaluated the role of CPS in bacteremia using a mouse model and in abortion using a pregnant guinea pig model following oral challenge. Compared with C. jejuni NCTC 11168 and 81-176, a clone SA isolate (IA3902) resulted in significantly higher bacterial counts and a significantly longer duration of bacteremia in mice. The loss of capsule production via gene-specific mutagenesis in IA3902 led to the complete abolishment of bacteremia in mice and abortion in pregnant guinea pigs, while complementation of capsule expression almost fully restored these phenotypes. The capsule mutant strain was also impaired for survival in guinea pig sera and sheep blood. Sequence-based analyses revealed that clone SA possesses a unique CPS locus with a mosaic structure, which has been stably maintained in all clone SA isolates derived from various hosts and times. These findings establish CPS as a key virulence factor for the induction of systemic infection and abortion in pregnant animals and provide a viable candidate for the development of vaccines against hypervirulent C. jejuni.


Population ecology of Heteronyx piceus (Coleoptera: Scarabaeidae) in a peanut/maize cropping system.
Large larval populations of the scarabaeid beetle Heteronyx piceus Blanchard that occur under peanuts, but not maize, in the South Burnett region of Australia are the result of a high rate and prolonged period of egg production by females feeding on peanut foliage. Heteronyx piceus is a relatively sedentary species and movement of females between adjacent fields is low. Populations of H. piceus varied markedly with landscape position. High larval populations are more likely (1 in 4 chance) to be encountered on the 'scrub' soils in the upper parts of the landscape than in the 'forest' soils in the lower half (1 in 20 chance), indicating that soil type/landscape position is a key risk factor in assessing the need for management intervention. The studies indicate that, because of the species' sedentary nature, the most meaningful population entity for management of H. piceus is the individual field, rather than the whole-farm or the region. The implications of this population ecology for management of the pest are discussed in relation to control strategies.


Medical student exposure to women's health concepts and practices: a content analysis of curriculum at Canadian medical schools.
Women's health (WH) includes a broad array of concerns and challenges that affect health across the lifespan. Considerable research shows that women continue to experience disparities in access to and quality of care. Apart from surveys of medical trainees and faculty, little research and none in Canada examined medical curriculum for WH. This study assessed how Canadian medical schools integrate WH in their curriculum. We used deductive and summative content analysis to describe instances and the nature of WH topics in program and course descriptions that were publicly-available on web sites of Canadian medical schools. We reported results using summary statistics and text examples. We employed a framework, tested in our prior research, that included mention of women's health principles and practices relevant to any health concern or condition including factors (e.g. sex, gender, social determinants) that influence health, and access to or quality of care. We retrieved 1459 documents from 16 medical schools (median 49.5, range 16 to 301). Few mentioned WH (125, 8.6 %), and the quantity of mentions varied by school (range 0.0-37.5 %). Pre-clerkship course documents more frequently mentioned WH (61/374, 17.3 %, chi square 43.2, p < 0.00001) compared with clerkship course documents (58/1067, 5.4 %). Core course documents more frequently mentioned WH (72/542, 13.3 %, chi square 29.0, p < 0.00001) compared with elective course documents WH (47/899, 5.2 %). Overall, documents more frequently referred to the WH domain of social determinants of health (88, 70.4 %). Few documents addressed women's health (21, 16.8 %), sex or gender (19, 15.2 %), other considerations (15.2 %) or principles/components of women's health (2, 1.6 %). Most documents that mentioned WH provided little detail about what those concepts referred to or how to optimize WH. Based on program and course descriptions, WH may not be well-integrated at Canadian medical schools, and future physicians may not be consistently exposed to the full breadth of WH. This reveals opportunities for enhancing WH in the medical curriculum. Future research is needed to engage stakeholders including women in developing, implementing and evaluating competencies and corresponding curriculum that reflect the full range of WH concepts and practices.


Isolation and characterization of a pock-forming plasmid pTA4001 from Streptomyces lavendulae.
A covalently closed circular (ccc) DNA with a size of 5.9 kb was isolated from a strain producing streptothricins , Streptomyces lavendulae No. 1080, and its restriction map was determined. Colonies of the plasmid-carrying strain formed pocks on the lawn of the plasmid-free derivatives of the same organism. The pock-forming ability of pTA4001 was confirmed by transformation of S. lividans with the plasmid DNA. Insertion of the chromosomal streptothricin resistance gene of S. lavendulae No. 1080 into pTA4001 gave various composite plasmids with marked deletion in S. lividans as a host. By analyzing these derivatives, a 2.2 kb region essential for replication and a possible region for pock formation were determined in the map of pTA4001 . A cloning vector, pKST2 (4.3 kb) containing resistance genes to streptothricin and thiostrepton with several unique cloning sites was constructed from pTA4001 .


Health disparities in staging of SEER-medicare prostate cancer patients in the United States.
To examine whether race or age disparities affected the odds of being staged among prostate cancer (PC) patients. Accurate staging is critical for determining treatment for PC. Multivariable logistic regression models examined race and age disparities with respect to the odds of being staged among PC patients using Surveillance, Epidemiology, and End Results-Medicare data. Similar analyses were performed to estimate the adjusted odds of being staged with distant metastatic vs in situ or local/regional disease. The proportion of patients without staging ranged between 3% and 16% by age and between 6% and 8% by race. Adjusted results demonstrated statistically significant lower odds ratios (P <.05) for 70-74, 75-79, and 80+-year-olds of 0.76, 0.52, and 0.23, respectively, relative to PC patients aged 65-69. The odds of being staged for African Americans are 0.78 times that of non-Hispanic Whites (95% confidence interval = 0.72-0.86). The adjusted probability of distant metastatic disease at initial diagnosis is higher for African Americans (odds ratio = 1.61; 95% confidence interval = 1.47-1.76) and older men with odds ratios of 1.25, 1.85, and 4.33 for ages 70-74, 75-79, and 80+, respectively, compared with 65-69-year-olds (all P <.05). Even though the overall odds of being staged increased over time, race and age disparities persisted. When staging did occur, the probability of distant metastatic disease was high for African Americans, and there were increasing odds of metastatic disease for all men with advanced age.


Risk of firearm injuries among children and youth of immigrant families.
Firearm injuries contribute to substantial morbidity and mortality. The immigrant paradox suggests that, despite being more socially disadvantaged, immigrants are less likely than nonimmigrants to have poor outcomes. We tested the association of immigrant characteristics with firearm injuries among children and youth. We conducted a population-based cohort study involving residents of Ontario aged 24 years and younger from 2008-2012 using health and administrative databases. We estimated rate ratios of unintentional and assault-related firearm injuries by immigrant status using Poisson regression models with Generalized Estimating Equations. We included 15 866 954 nonimmigrant and 4 551 291 immigrant person-years in our analysis. Nonimmigrant males had 1032 unintentional (12.4 per 100 000, 95% confidence interval [CI] 11.7-13.2) and 304 assault-related (3.6 per 100 000, 95% CI 3.2-4.0) firearm injuries. Immigrant males had 148 unintentional (7.2 per 100 000, 95% CI 6.1-8.5) and 113 assault-related (5.5 per 100 000, 95% CI 4.5-6.6) firearm injuries. Compared with nonimmigrants, immigrants had a lower rate of unintentional firearm injury (adjusted rate ratio 0.5, 95% CI 0.4-0.6) but a similar rate of assault-related firearm injury. Among immigrants, refugees had a 43% higher risk of assault-related firearm injury compared with nonrefugees (adjusted rate ratio 1.4, 95% CI 1.0-2.0). Immigrants from Central America and Africa accounted for 68% of immigrants with assault-related firearm injuries. Compared with nonimmigrants, immigrant children and youth had a lower risk of unintentional firearm injury, although the risk of assault-related firearm injury was higher among refugees and immigrants from Central America and Africa. The results suggest that prevention strategies for firearm safety should target nonimmigrant youth as well as these newly identified high-risk immigrant populations.


Evaluation of thoracic limb loads, elbow movement, and morphology in dogs before and after arthroscopic management of unilateral medial coronoid process disease.
To (1) evaluate thoracic limb loads and symmetry, and elbow function and morphology, before and after arthroscopic treatment of unilateral medial coronoid process disease (MCPD), and (2) determine if functional variables correlate with morphologic findings. Prospective case series. Dogs (n = 14) with thoracic limb lameness. Dogs were included when unilateral MCPD was confirmed as the cause of lameness. Kinetic analysis of both thoracic limbs, along with kinematic analysis and goniometry of both elbows were carried out before, and 60, 120, and 180 days after partial coronoidectomy by arthroscopy. Radiography and computed tomography of both elbows were performed before and 180 days after arthroscopy. A nonsignificant (P = .11) increase in the peak vertical loads (PFz), and a significant (P = .022) increase in the vertical impulse (iFz) applied by the affected limb were seen. Symmetry indices improved, with significant differences between sessions (PFz: P = .019; iFz: P = .003). Kinematic variables showed no significant differences, between sessions or when comparing both elbows within sessions. Goniometry revealed no significant differences between sessions, but some significant differences were identified when comparing both elbows within sessions. Osteophytosis and degree of lameness showed no correlation, before (rs = -0.077; P = .79) or after arthroscopy (rs = 0.27; P = .35). Kinetic variables improved after arthroscopy, without full restoration of function. Kinematic variables did not change significantly. Osteoarthritis and goniometric measurements in the affected joint worsened. Functional variables did not correlate with morphologic findings.


Sleep and screen exposure across the beginning of life: deciphering the links using big-data analytics.
Evidence for the association between screen time and insufficient sleep is bourgeoning, and recent findings suggest that these associations may be more pronounced in younger compared to older children, and for portable compared to non-portable devices. However, these effects have yet to be investigated within the beginning of life. Importantly, there are no data for the relationship between screen exposure and objectively measured infant sleep. This study examined the moderating role of age for both touchscreens' and television's relationship with sleep, using auto-videosomnography within a big-data sample of infants. The sleep of 1074 infants (46% girls) aged 0-18 months was objectively assessed using computer-vision technology in this cross-sectional study. Sleep was additionally reported by parents in an online survey, as was infant exposure to screens. Age significantly moderated the relationship between daytime touchscreen exposure and sleep with a distinct pattern for younger infants, in which screen exposure was associated with decreased daytime sleep, but with a proposed compensatory increase in nighttime sleep consolidation. Compared to touchscreens, television exposure was less likely to be associated with sleep metrics, and age moderated this relationship only for daytime and 24-hour sleep duration. In young infants, a daytime-nighttime sleep "trade-off" emerged, suggesting that the displacement of daytime sleep by screens may lead to greater accumulation of sleep homeostatic pressure, which in turn facilitates more consolidated nighttime sleep.


Adenosine-induced presynaptic inhibition of IPSCs and EPSCs in rat hypothalamic supraoptic nucleus neurones.
1. The effects of adenosine on synaptic transmission in magnocellular neurosecretory cells were investigated using whole-cell patch-clamp recordings in acute rat hypothalamic slices that included the supraoptic nucleus. 2. Adenosine reversibly reduced the amplitude of evoked inhibitory (IPSCs) and excitatory (EPSCs) postsynaptic currents in a dose-dependent manner (IC50 approximately 10 microM for both types of current). 3. Depression of IPSCs and EPSCs by adenosine was reversed by the application of the A1 adenosine receptor antagonist 8-cyclopentyl-1, 3-dimethylxanthine (CPT; 10 microM). 4. When pairs of stimuli were given at short intervals, adenosine inhibitory action was always less effective on the second of the two responses than on the first, resulting in an increased paired-pulse facilitation and suggesting a presynaptic site of action. This observation was confirmed by analysis of spontaneous miniature synaptic currents whose frequency, but not amplitude or kinetics, was reversibly reduced by 100 microM adenosine. 5. CPT had no effect on synaptic responses evoked at a low frequency of stimulation (0.05-0.5 Hz), indicating the absence of tonic activation of A1 receptors under these recording conditions. However, CPT inhibited a time-dependent depression of both IPSCs and EPSCs induced during a 1 Hz train of stimuli. 6. Taken together, these results suggest that adenosine can be released within the supraoptic nucleus at a concentration sufficient to inhibit the release of GABA and glutamate via the activation of presynaptic A1 receptors. By its inhibitory feedback action on the major afferent inputs to oxytocin and vasopressin neurones, adenosine could optimally adjust electrical and secretory activities of hypothalamic magnocellular neurones.


[Some data from studying and using aminoglycoside antibiotics in reconstructive surgery].
Effectiveness of antibacterial drugs from the group of aminoglycoside antibiotics,i.e. kanamycin, gentamycin and tobramycin with respect to the main causative agents of surgical infections, i.e. Staphylococcus, Coli bacteria, Proteus and Ps. aeruginosa was shown. The highest activity against Ps. aeruginosa was registered to tobramycin. The aminoglycoside antibiotics were satisfactorily absorbed after their intramuscular administration providing therapeutic levels of the antibiotics in the blood. For instance, administration of gentamycin in a dose of 80 mg and tobramycin in a dose of 50 mg 1-2 times a day provided the blood levels higher than the minimun inhibitory concentrations for most strains of gramnegative flora including Ps. aeruginosa. The use of the above drugs in theraphy and prophylaxis of infections caused by Ps. aeruginosa is expedient.