The Wheat Wall-Associated Receptor-Like Kinase TaWAK-6D Mediates Broad Resistance to Two Fungal Pathogens Fusarium pseudograminearum and Rhizoctonia cerealis.
The soil-borne fungi Fusarium pseudograminearum and Rhizoctonia cerealis are the major pathogens for the economically important diseases Fusarium crown rot (FCR) and sharp eyespot of common wheat (Triticum aestivum), respectively. However, there has been no report on the broad resistance of wheat genes against both F. pseudograminearum and R. cerealis. In the current study, we identified TaWAK-6D, a wall-associated kinase (WAK) which is an encoding gene located on chromosome 6D, and demonstrated its broad resistance role in the wheat responses to both F. pseudograminearum and R. cerealis infection. TaWAK-6D transcript induction by F. pseudograminearum and R. cerealis was related to the resistance degree of wheat and the gene expression was significantly induced by exogenous pectin treatment. Silencing of TaWAK-6D compromised wheat resistance to F. pseudograminearum and R. cerealis, and repressed the expression of a serial of wheat defense-related genes. Ectopic expression of TaWAK-6D in Nicotiana benthamiana positively modulated the expression of several defense-related genes. TaWAK-6D protein was determined to localize to the plasma membrane in wheat and N. benthamiana. Collectively, the TaWAK-6D at the plasma membrane mediated the broad resistance responses to both F. pseudograminearum and R. cerealis in wheat at the seedling stage. This study, therefore, concludes that TaWAK-6D is a promising gene for improving wheat broad resistance to FCR and sharp eyespot.


Globular protein stabilized nanoparticles for delivery of disulfiram: fabrication, characterization, in vitro toxicity, and cellular uptake.
Disulfiram (DSF), an FDA-approved anti-alcoholic drug, has recently shown that it possesses anti-cancer effects. However, DSF is hydrophobic in nature with less stability. Therefore, new approaches are required for the effective delivery of DSF to treat cancers. Herein, we prepared DSF loaded soy protein isolate (SPI) nanosuspension (Ns) for enhancing the anti-cancer delivery of DSF. The optimized DSF-SPI-Ns had an average particle size of 164.28 ± 2.07 nm with a narrow size distribution of 0.217 ± 0.035 and zeta potential around -22.30 ± 2.11 mV, respectively. The highest drug loading and entrapment efficiency achieved was 5.516 ± 1.98%, and 91.61 ± 1.15%, respectively. The surface morphology of Ns was revealed by TEM, and the FTIR DSC, PXRD, and TGA were used for physicochemical characterization. Further, fluorescence spectroscopy and molecular docking studies were carried out to understand the interactions between (SPI and DSF) and binding sites of DSF on the surface of SPI, respectively. In vitro release studies showed a sustained release pattern and followed a Fickian diffusion release from the Ns. The in vitro cytotoxicity of SPI indicated the excellent biocompatibility, and DSF-SPI-Ns were found to be more cytotoxic compared to the free DSF solution. Moreover, the cellular uptake studies also indicated the effective delivery of the formulation to the cancer cells. Results of the current study suggested that the SPI coated Ns might be a promising drug delivery system for hydrophobic DSF, and the potential application of SPI as a coating/stabilizing agent for the delivery of hydrophobic/hydrophilic cancer therapeutics.


Clonidine for the Treatment of Agitation After Dexmedetomidine Discontinuation in Pediatric Patients: A Retrospective Cohort Study.
Dexmedetomidine has become a widely used drug in PICUs for sedation. We aim to determine the effects of clonidine on pediatric patients after dexmedetomidine use. This was a retrospective cohort study that evaluated all pediatric patients admitted to a tertiary PICU who received dexmedetomidine infusion for >48 hours. Outcomes in patients exposed to clonidine (CLON) were compared with those of patients who were not exposed (NoCLON). A total of 216 patients were included in this study (43 CLON and 173 NoCLON). The primary outcome, agitation, was less in the CLON cohort (9.3%) than in the NoCLON cohort (9.3% versus 29.5%, respectively; p < 0.01). Hospital LOS was longer in the CLON group (59 versus 20 days, p < 0.01), as was PICU LOS (37.4 versus 11.1 days, p < 0.01). There was no significant difference in the occurrence of increased heart rate or blood pressure between the 2 cohorts. Patients exposed to concurrent midazolam and opioid infusions had higher incidence of agitation when they did not receive clonidine (CLON 8% versus NoCLON 37%, OR 0.15; 95% CI, 0.05-0.51; p < 0.01). In contrast, there was no difference in the incidence of agitation for the CLON group versus the NoCLON group when dexmedetomidine was administered alone (25% versus 19%, OR 1.4; p = 0.99). Our study confirms the importance and effectiveness of clonidine to treat agitation after dexmedetomidine discontinuation. A validated withdrawal scoring tool can help better define dexmedetomidine withdrawal in pediatric patients.


Activation-dependent properties of pregnenolone sulfate inhibition of GABAA receptor-mediated current.
Sulfated steroids like pregnenolone sulfate (PS) are found endogenously in the central nervous system where they may modulate GABAA receptors. Understanding the mechanism of steroid inhibition is important for understanding the conditions under which endogenous steroids modulate GABAA receptor function, assessing their potential clinical utility, and for evaluating sulfated steroids as probes of receptor behaviour. Some previous studies suggest that sulfated steroid inhibition exhibits activation dependence, whilst other studies suggest only slow, time-dependent inhibition, perhaps reflecting slow PS association with receptors. We tested activation dependence in several ways. Steroid potency increased 2- to 3-fold with approximately 10-fold change in GABA concentration. PS inhibition of saturating partial agonist responses suggested that the level of channel activation, rather than receptor occupancy by agonist, is important for PS inhibition. Inhibition by sulfated steroids exhibited weak or no voltage dependence. Responses to rapid applications of exogenous GABA differed little whether PS was pre-applied or simply co-applied with GABA, consistent with the hypothesis that the actions of PS are facilitated by receptor activation. PS applied during steady-state GABA responses exhibited slow onset and offset rate constants. The offset, rather than onset, was significantly slowed by elevated GABA concentration. At hippocampal synapses, large, multiquantal IPSCs were inhibited more effectively by a fixed concentration of PS than small quantal content IPSCs, consistent with known 'pooling' of transmitter following multiquantal release. Picrotoxinin, although superficially similar to PS in its activation dependence, was dissimilar from PS in a number of details. In summary, PS inhibition exhibits activation dependence that may be explained by activation-dependent binding and altered desensitization.


Communication: papillomavirus DNA in basal cell carcinomas of immunocompetent patients: an accidental association?TITLE.
DNA of human papillomaviruses has frequently been detected in nonmelanoma skin cancers, raising the question of a possible causal contribution of these tumor viruses to skin carcinogenesis. Basal cell carcinomas are the most common nonmelanoma skin cancers; however, so far they are only poorly analyzed with regard to human papillomavirus infection. We searched for human papillomavirus-DNA in 69 biopsies from 61 immunocompetent basal cell carcinoma patients from two geographic locations in Europe using six different polymerase chain reaction primer systems. We could demonstrate human papillomavirus-DNA in 43.5% of the tested tumors. Human papillomavirus positivity did not seem to correlate with the duration of disease or patients' age. The vast majority of virus types in the biopsies belonged to the group of epidermodysplasia verruciformis-associated human papillomavirus. Of 31 sample pairs tested for human papillomavirus-DNA in tumors as well as in perilesional healthy skin, seven carried viral sequences in lesional and healthy skin and three only in the basal cell carcinoma. Six of the seven human papillomavirus-positive basal cell carcinoma/healthy skin pairs contained identical human papillomavirus types in tumors and histologically normal tissue. Forty basal cell carcinoma patients were additionally analyzed for IgG antibodies against virus-like particles of three representative epidermodysplasia verruciformis-human papillomavirus types: 8, 15, and 36. No statistically significant differences could be detected between human papillomavirus antibody prevalences of basal cell carcinoma patients and of dermatologically healthy individuals. Moreover, serologic findings did not correlate with the detection of specific human papillomavirus types in tumors. Our results seem to suggest that the occurrence of human papillomavirus-DNA in basal cell carcinoma does not reflect a major etiologic role of human papillomavirus in this cancer.


Toxoplasma gondii glycosylphosphatidylinositols up-regulate major histocompatibility complex (MHC) molecule expression on primary murine macrophages.
Toxoplasma gondii is an obligatory intracellular parasite able to block the IFN-gamma-induced up-regulation of major histocompatibility complex (MHC) class I and class II molecules. This facilitates parasite-mediated evasion of T-cell responses. Glycosylphosphatidylinositols (GPIs) are involved in the pathogenicity of protozoan parasites and we investigated if GPIs are responsible for inhibition of MHC expression on macrophages. In contrast to the blockade observed in cells infected with viable tachyzoites, T. gondii GPIs up-regulated MHC class I and class II molecules on the surface of both unstimulated and IFN-gamma-stimulated primary murine macrophages. This effect was correlated to the ability of GPIs to increase the antigen presentation to CD8(+) lymphocytes. T. gondii GPIs did not activate STAT1, one of the factors involved in the transcription of MHC class I and class II genes. However, the GPI-induced MHC class I up-regulation was abrogated by SN50, a specific NF-KB inhibitor. Up-regulation of surface MHC molecules by GPIs may lead to the elimination of non-infected cells of the host immune system, contributing to the immune escape strategy of T. gondii.


Effects of antihistamines on the cardiopulmonary changes due to canine anaphylaxis.
Histamine has long been considered to be an important chemical mediator in the pathogenesis of immediate hypersensitivity reactions. We evaluated the efficacy of antihistamines to determine the physiological role of histamine in canine anaphylaxis. Either a saline vehicle (control group), an H1 antihistamine (chlorpheniramine, 10 mg/kg), or this H1 antihistamine and an H2 antihistamine (cimetidine, 30 mg/kg) was administered to three separate groups of anesthetized dogs (n = 8). Cardiopulmonary responses and plasma histamine levels were measured after the separate intravenous injection of Ascaris suum antigen and histamine. Results were analyzed only from the animals demonstrating physiological responses or histamine release after antigen injection. In the control group, antigen produced a 43 +/- 15% (mean +/- SE) decrease in mean arterial blood pressure, a 34 +/- 13% fall in cardiac output, and a 19 +/- 9% decrease in lung compliance, whereas pulmonary vascular resistance increased 161 +/- 87% and airway resistance rose 114 +/- 66%. Similar physiological abnormalities were observed with histamine shock. However, peak plasma histamine levels were, in most cases, greater after histamine injection than after antigen injection. An H1 antihistamine alone or in combination with an H2 antihistamine did not alter the physiological changes associated with systemic anaphylaxis. In contrast, the combined use of H1 and H2 antihistamines prevented the cardiopulmonary responses associated with the intravenous administration of histamine.(ABSTRACT TRUNCATED AT 250 WORDS)


Physical activity of German children during different segments of the school day.
This study objectively investigated the amount and intensity of German primary school children's physical activity (PA) during different segments of the school day and explored the contribution of physical education (PE) and break times to daily moderate to vigorous PA (MVPA). PA of 294 children (7.1 ± 0.7 years, 48 % male) was objectively measured for 6 days using Actiheart®. Based on children's timetables, break times and PE periods were determined and PA was calculated individually and subsequently classified in light (1.5-3 MET), moderate (3-6 MET) and vigorous (>6 MET) intensities. Weight status was determined during a school visit. Children spent 133 ± 61 min in MVPA; on weekdays, this amount increased significantly (141 ± 66 min, p ≤ 0.01). 45.9 % of children reached physical activity guidelines of 60 min of MVPA daily, with boys achieving this goal significantly more often than girls (65.6 vs. 28.7 %, respectively; p ≤ 0.01). PE lessons and break times accounted for 15 ± 13 min (12.7 %) and 7 ± 6 min (5.8 %) of daily MVPA, respectively. On days with PE, children spent 144 ± 68 min in MVPA, whereas on days without PE, this time decreased significantly to 122 ± 63 min (p ≤ 0.01). The findings suggest that segments such as PE lessons and morning breaks are important sources for MVPA for boys and girls. This should therefore be considered for policies, timetables and curriculums in order to offer sufficient opportunities for children to be physically active during the school day.


Aquaporin-4 in the central nervous system: cellular and subcellular distribution and coexpression with KIR4.1.
Aquaporin-4 (AQP4) is the predominant water channel in the neuropil of the central nervous system. It is expressed primarily in astrocytes, but also occurs in ependymocytes and endothelial cells. A striking feature of AQP4 expression is its polarized distribution in brain astrocytes and retinal Muller cells. Thus, immunogold analyses have revealed an enrichment of AQP4 in endfeet membranes in contact with brain microvessels or subarachnoidal space and a low but significant concentration in non-endfeet membranes, including those astrocyte membranes that ensheath glutamate synapses. The subcellular compartmentation of AQP4 mimics that of the potassium channel Kir4.1, which is implicated in spatial buffering of K(+). We propose that AQP4 works in concert with Kir4.1 and the electrogenic bicarbonate transporter NBC and that water flux through AQP4 contributes to the activity dependent volume changes of the extracellular space. Such volume changes are important as they affect the extracellular solute concentrations and electrical fields, and hence neuronal excitability. We conclude that AQP4-mediated water flux represents an integral element of brain volume and ion homeostasis.


Generation of a CRISPR database for Yersinia pseudotuberculosis complex and role of CRISPR-based immunity in conjugation.
The clustered regularly interspaced short palindromic repeat - CRISPR-associated genes (CRISPR-Cas) system is used by bacteria and archaea against invading conjugative plasmids or bacteriophages. Central to this immunity system are genomic CRISPR loci that contain fragments of invading DNA. These are maintained as spacers in the CRISPR loci between direct repeats and the spacer composition in any bacterium reflects its evolutionary history. We analysed the CRISPR locus sequences of 335 Yersinia pseudotuberculosis complex strains. Altogether 1902 different spacer sequences were identified and these were used to generate a database for the spacer sequences. Only ∼10% of the spacer sequences found matching sequences. In addition, surprisingly few spacers were shared by Yersinia pestis and Y. pseudotuberculosis strains. Interestingly, 32 different protospacers were present in the conjugative plasmid pYptb32953. The corresponding spacers were identified from 35 different Y. pseudotuberculosis strains indicating that these strains had encountered pYptb32953 earlier. In conjugation experiments, pYptb32953-specific spacers generally prevented conjugation with spacer-positive and spacer-free strains. However, some strains with one to four spacers were invaded by pYptb32953 and some spacer-free strains were fully resistant. Also some spacer-positive strains were intermediate resistant to conjugation. This suggests that one or more other defence systems are determining conjugation efficiency independent of the CRISPR-Cas system.


When parasites disagree: evidence for parasite-induced sabotage of host manipulation.
Host manipulation is a common parasite strategy to alter host behavior in a manner to enhance parasite fitness usually by increasing the parasite's transmission to the next host. In nature, hosts often harbor multiple parasites with agreeing or conflicting interests over host manipulation. Natural selection might drive such parasites to cooperation, compromise, or sabotage. Sabotage would occur if one parasite suppresses the manipulation of another. Experimental studies on the effect of multi-parasite interactions on host manipulation are scarce, clear experimental evidence for sabotage is elusive. We tested the effect of multiple infections on host manipulation using laboratory-bred copepods experimentally infected with the trophically transmitted tapeworm Schistocephalus solidus. This parasite is known to manipulate its host depending on its own developmental stage. Coinfecting parasites with the same aim enhance each other's manipulation but only after reaching infectivity. If the coinfecting parasites disagree over host manipulation, the infective parasite wins this conflict: the noninfective one has no effect. The winning (i.e., infective) parasite suppresses the manipulation of its noninfective competitor. This presents conclusive experimental evidence for both cooperation in and sabotage of host manipulation and hence a proof of principal that one parasite can alter and even neutralize manipulation by another.


Synchronization and extinction in cyclic games with mixed strategies.
We consider cyclic Lotka-Volterra models with three and four strategies where at every interaction agents play a strategy using a time-dependent probability distribution. Agents learn from a loss by reducing the probability to play a losing strategy at the next interaction. For that, an agent is described as an urn containing β balls of three and four types, respectively, where after a loss one of the balls corresponding to the losing strategy is replaced by a ball representing the winning strategy. Using both mean-field rate equations and numerical simulations, we investigate a range of quantities that allows us to characterize the properties of these cyclic models with time-dependent probability distributions. For the three-strategy case in a spatial setting we observe a transition from neutrally stable to stable when changing the level of discretization of the probability distribution. For large values of β, yielding a good approximation to a continuous distribution, spatially synchronized temporal oscillations dominate the system. For the four-strategy game the system is always neutrally stable, but different regimes emerge, depending on the size of the system and the level of discretization.


The recomBead Borrelia antibody index, CXCL13 and total IgM index for laboratory diagnosis of Lyme neuroborreliosis in children.
For laboratory diagnostics of Lyme neuroborreliosis (LNB), the recomBead Borrelia antibody index (AI) assay has shown promising results in a mixed age population, but has not previously been evaluated with specific focus on paediatric patients. The aim of the study was to evaluate the recomBead Borrelia AI assay in cerebrospinal fluid (CSF) for the laboratory diagnosis of LNB in children. We also wanted to explore whether early markers, such as CXCL13 in CSF and/or total IgM index could be useful as complementary diagnostic tools. Children being evaluated for LNB in a Swedish Lyme endemic area were included in the study (n = 146). Serum and CSF were collected on admission. Patients with other specific diagnoses were controls (n = 15). The recomBead Borrelia AI assay and the recomBead CXCL13 assay (Mikrogen) were applied together with total IgM index. The overall sensitivity for recomBead Borrelia AI (IgM and IgG together) was 74% and the specificity was 97%. However, the highest sensitivity (91%) at an acceptable level of specificity (90%) was obtained by recomBead Borrelia AI together with CXCL13 and total IgM index, showing a positive predictive value of 84% and a negative predictive value of 95%. Thus, the recomBead Borrelia AI assay performs with moderate sensitivity and high specificity in paediatric LNB patients. The major advantage seems to be increased sensitivity in the possible LNB group compared to the IDEIA assay. The diagnostic sensitivity may be further increased by using a combination of early markers, such as CXCL13 in CSF and total IgM index.


Dorsomedial cutaneous nerve syndrome: treatment with nerve transection and burial into bone.
Damage to the dorsomedial cutaneous nerve of the foot, which innervates the medial hallux, may occur with crush injury or iatrogenically with bunion surgery. Severe neuritic pain after bunion surgery may alert the surgeon that this small nerve has been damaged. The term "dorsomedial cutaneous nerve syndrome" is suggested for this condition, and nine patients with such forefoot presentations, all of which were unresponsive to nonoperative interventions, are described. The nerve had been either transected or bound in scar tissue; in these nine cases, the nerve was then resected and buried in the proximal aspect of the first metatarsal or the medial cuneiform. Most patients underwent an additional procedure (other than the nerve procedure), such as revision bunionectomy or arthrodesis, but all felt they could clearly delineate nerve pain from bone or joint pain. All patients experienced marked relief of their symptoms, usually within days after the surgery, and were satisfied with the results. The verbal analog pain score, on a scale of 0 (no pain) to 10 (pain requiring amputation), improved from a preoperative level of 8.6 to a postoperative level of 2.0. Resection and burial of this nerve appears to be a useful treatment for neuritis unresponsive to nonoperative measures.


Protein restriction cycles reduce IGF-1 and phosphorylated Tau, and improve behavioral performance in an Alzheimer's disease mouse model.
In laboratory animals, calorie restriction (CR) protects against aging, oxidative stress, and neurodegenerative pathologies. Reduced levels of growth hormone and IGF-1, which mediate some of the protective effects of CR, can also extend longevity and/or protect against age-related diseases in rodents and humans. However, severely restricted diets are difficult to maintain and are associated with chronically low weight and other major side effects. Here we show that 4 months of periodic protein restriction cycles (PRCs) with supplementation of nonessential amino acids in mice already displaying significant cognitive impairment and Alzheimer's disease (AD)-like pathology reduced circulating IGF-1 levels by 30-70% and caused an 8-fold increase in IGFBP-1. Whereas PRCs did not affect the levels of β amyloid (Aβ), they decreased tau phosphorylation in the hippocampus and alleviated the age-dependent impairment in cognitive performance. These results indicate that periodic protein restriction cycles without CR can promote changes in circulating growth factors and tau phosphorylation associated with protection against age-related neuropathologies.


Pathomechanisms of Altered Wound Healing in Recessive Dystrophic Epidermolysis Bullosa.
Individuals with recessive dystrophic epidermolysis bullosa (RDEB), a rare genetic skin disease, carry mutations in the COL7A1 gene that codes for type VII collagen, an extracellular matrix component of the basement membrane zone forming the anchoring fibrils. As a consequence, RDEB individuals manifest unremitting skin blistering that evolves into chronic wounds, inflammation, and fibrosis. These features play a central role in the development of more severe disease complications, such as mitten deformities of hands and feet and aggressive epithelial cancers. Despite being recognized as a central clinical issue for RDEB, wound healing impairment has been only marginally investigated. Recently, studies with disease mouse models started to shed light on the molecular mechanisms underlying the altered healing response of RDEB. In turn, alterations found in RDEB skin cell behavior fostered the understanding of mechanisms that may be responsible for defective skin repair. This review summarizes findings related to healing impairment in RDEB, and highlights therapeutic strategies for ameliorating healing.


E2F1 mediated apoptosis induced by the DNA damage response is blocked by EBV nuclear antigen 3C in lymphoblastoid cells.
EBV latent antigen EBNA3C is indispensible for in vitro B-cell immortalization resulting in continuously proliferating lymphoblastoid cell lines (LCLs). EBNA3C was previously shown to target pRb for ubiquitin-proteasome mediated degradation, which facilitates G1 to S transition controlled by the major transcriptional activator E2F1. E2F1 also plays a pivotal role in regulating DNA damage induced apoptosis through both p53-dependent and -independent pathways. In this study, we demonstrate that in response to DNA damage LCLs knocked down for EBNA3C undergo a drastic induction of apoptosis, as a possible consequence of both p53- and E2F1-mediated activities. Importantly, EBNA3C was previously shown to suppress p53-induced apoptosis. Now, we also show that EBNA3C efficiently blocks E2F1-mediated apoptosis, as well as its anti-proliferative effects in a p53-independent manner, in response to DNA damage. The N- and C-terminal domains of EBNA3C form a stable pRb independent complex with the N-terminal DNA-binding region of E2F1 responsible for inducing apoptosis. Mechanistically, we show that EBNA3C represses E2F1 transcriptional activity via blocking its DNA-binding activity at the responsive promoters of p73 and Apaf-1 apoptosis induced genes, and also facilitates E2F1 degradation in an ubiquitin-proteasome dependent fashion. Moreover, in response to DNA damage, E2F1 knockdown LCLs exhibited a significant reduction in apoptosis with higher cell-viability. In the presence of normal mitogenic stimuli the growth rate of LCLs knockdown for E2F1 was markedly impaired; indicating that E2F1 plays a dual role in EBV positive cells and that active engagement of the EBNA3C-E2F1 complex is crucial for inhibition of DNA damage induced E2F1-mediated apoptosis. This study offers novel insights into our current understanding of EBV biology and enhances the potential for development of effective therapies against EBV associated B-cell lymphomas.


[Age determination of venous thrombi by ultrasound].
In addition to diagnosing deep-vein thrombosis of the lower limb and pelvis, determination of the age of the thrombus is of considerable importance in guiding treatment. The present study was done to find out whether thrombus age can be determined by ultrasound morphology. 138 examinations were carried out on 76 lower limbs with phlebographically demonstrated thrombosed veins. Diameter of the thrombosed vein, echogenicity of the thrombus and demarcation of the venous wall from its surroundings were determined by colour-coded duplex sonography. The results were compared with histological evidence or with the age of the thrombus as determined by history and clinical course. The diameter of the thrombosed vein in conjunction with the history and clinical data makes it possible to determine the age of a thrombus. Echogenicity of the thrombus and demarcation of the venous wall do not provide any reliable information on thrombus age.


Shutting down secondary reaction pathways: the essential role of the pyrrolyl ligand in improving silica supported d(0)-ML4 alkene metathesis catalysts from DFT calculations.
The efficiency of silica supported d(0) ML(4) alkene metathesis catalysts [([triple bond]SiO)M(NR(1))(=CHR(2))(X)] (M = Mo, W; R(1) = aryl and alkyl) is influenced by the nature of the X ancillary ligand. Replacing the alkyl ligand by a pyrrolyl ligand dramatically increases the performance of the catalyst. DFT calculations on the metathesis, the deactivation, and the byproduct formation pathways for the imido Mo and W and the alkylidyne Re complexes give a rational for the role of pyrrolyl ligand. Dissymmetry at the metal center leads to more efficient catalyst even when the difference in sigma-donating ability between X and OSi is not large. beta-H transfer at the square based pyramid metallacyclobutane is the key step for catalyst deactivation and byproduct formation. Overall, the greatest benefit of substituting the ancillary alkyl by a pyrrolyl ligand, [([triple bond]SiO)M(ER(1))(=CHR(2))(pyrrolyl)], is in fact not to improve the efficiency of the catalytic cycle of alkene metathesis, but to shut down deactivation and byproduct formation pathways. Pyrrolyl ligand, and more generally ligands having metal-bound-atoms more electronegative than carbon, disfavor mostly the two first steps (beta-H transfer at the metallacyclobutane and subsequent insertion of an ethene in the M-H bond) of the deactivation channel. The [([triple bond]SiO)M(ER(1))(=CHR(2))(pyrrolyl)] catalyst is thus highly efficient because pyrrolyl ligand is optimal: (i) it is still a better electron donor than the siloxy group, thus, favoring the metathesis pathway (dissymmetry at the metal center); and (ii) the nitrogen of the pyrrolyl ligand is more electronegative than the carbon of the alkyl group, thus, specifically disfavoring the decomposition of the metallacyclobutane intermediate via beta-H transfer.


Phorbol ester receptors: autoradiographic identification in the developing rat.
Autoradiography with 3H-labeled phorbol dibutyrate was used for the light microscopic detection of phorbol ester receptors in rat fetuses. In 15- and 18-day fetuses, as well as in adult rats, receptors were found to be concentrated in the central nervous system. The localization of receptors in the ventral marginal zone of the fetal neural tube, the lens of the eye, and other sites suggests a role for phorbol ester receptors in cellular process extension and cell-cell interaction.


Clinical clues predictive of Stevens-Johnson syndrome as the cause of chronic cicatrising conjunctivitis.
This study aimed to identify the clinical clues in patients with chronic cicatrising conjunctivitis (CCC), that were suggestive of Stevens-Johnson syndrome (SJS) as the aetiology. This was a cross-sectional observational study of 75 patients presenting with CCC from 2016 to 2018. Those with a documented diagnosis of SJS (n=43) were included as cases; while those with a positive serology or tissue biopsy for a non-SJS condition were included as controls (n=32). The features in the medical history and clinical examination that were positively and negatively associated with SJS were scored +1 and -1, respectively. A receiver operating characteristic (ROC) curve analysis was performed to detect the threshold score for optimal sensitivity and specificity of the scoring system. No single feature had absolute sensitivity and specify for SJS. The 10 positive features suggestive of SJS (p<0.0001) included (1) history of: acute conjunctivitis, fever or drug intake preceding conjunctivitis, peeling of skin on pressure, loss of nails and severe morbidity with hospital admission; and (2) clinical features of: skin discoloration, nail disfigurement, lip-margin dermalisation, lid-margin keratinisation and distichiasis. The two negative criteria were history of mucosal ulcers without skin involvement and recurrent mucosal ulceration. On ROC analysis, a score of >5 showed a sensitivity of 90.7% and specificity of 93.8% for the diagnosis of SJS. The combination of clinical clues identified in this study can help clinicians confirm SJS as the aetiology of conjunctival cicatrisation, especially when reliable documentation of the acute episode is not available.


Effect of Angiotensin(1-7) on Heart Function in an Experimental Rat Model of Obesity.
Obesity is a risk factor for the development of cardiovascular diseases. Recently it was shown that overexpression of the Mas-receptor antagonist angiotensin(1-7) could prevent from diet-induced obesity. However, it remained unclear whether diet-induced obesity and angiotensin(1-7) overexpression might also have effects on the cardiovascular system in these rats. Twenty three male Sprague Dawley rats were fed with standard chow (SD+chow, n = 5) or a cafeteria diet (SD+CD, n = 6) for 5 months. To investigate the effect of angiotensin(1-7) transgenic rats, expressing an angiotensin(1-7)-producing fusion protein in testis were used. These transgenic rats also received a 5 month's feeding period with either chow (TGR+chow, n = 6) or cafeteria diet (TGR+CD, n = 6), respectively. Hemodynamic measurements (pressure-volume loops) were carried out to assess cardiac function and blood pressure. Subsequently, hearts were explanted and investigated according to the Langendorff technique. Furthermore, cardiac remodeling in these animals was investigated histologically. After 5 months cafeteria diet feeding rats showed a significantly increased body weight, which could be prevented in transgenic rats. However, there was no effect on cardiac performance after cafeteria diet in non-transgenic and transgenic rats. Moreover, overexpression of angiotensin(1-7) deteriorated cardiac contractility as indicated by impaired dp/dt. Furthermore, histological analysis revealed that cafeteria diet led to myocardial fibrosis in both, control and transgenic rats and this was not inhibited by an overproduction of angiotensin(1-7). These results indicate that an overexpression of circulating angiotensin(1-7) prevents a cafeteria diet-induced increase in body weight, but does not affect cardiac performance in this experimental rat model of obesity. Furthermore, overexpression of angiotensin(1-7) alone resulted in an impairment of cardiac function.


Influence of heart rate and atrial transport on left ventricular volume and function: relation to hemodynamic changes produced by supraventricular arrhythmia.
The response of the left ventricle to pacing-induced changes in heart rate and the atrioventricular (A-V) relation was examined with equilibrium gated radionuclide ventriculography in 20 patients who had normal ventricular function after surgery for recurrent supraventricular tachycardia. In 10 patients count-derived left ventricular ejection fraction, end-diastolic volume and stroke volume were measured during sinus rhythm and during atrial pacing at 120, 140 and 160 beats/min. In the other 10 patients similar determinations were made during sequential A-V and simultaneous ventricular and atrial (V/A) pacing, both at rates of 100 and 160 beats/min. Left ventricular ejection fraction did not change significantly with atrial pacing (from 0.65 +/- 0.02 [mean +/- standard error of the mean] at a baseline sinus rate of 91 +/- 3 beats/min to 0.62 +/- 0.03 at 160 beats/min) despite a progressive decrease in end-diastolic volume. The percent reduction in end-diastolic volume (% delta EDV) and stroke volume (+ delta SV) from the baseline values was linear and related to change in heart rate (delta HR) as % delta EDV = -0.60 delta HR + 5.19 (r = 0.71; p less than 0.01) and % delta SV = -0.62 delta HR + 5.03 (r = 0.76; p less than 0.001). Left ventricular ejection fraction with baseline sequential A-V pacing at 100 beats/min was 0.67 +/- 0.03 and not significantly altered by either sequential A-V or simultaneous V/A pacing at 160 beats/min. At 100 beats/min, loss of atrial transport with simultaneous V/A pacing resulted in a small reduction in end-diastolic volume from a baseline value of -9.0 +/- 1.9 percent (p less than 0.01) and a nonsignificant reduction in stroke volume of -3.7 +/- 1.6 percent. During simultaneous V/A pacing at 160 beats/min, the reduction in end-diastolic and stroke volumes from the baseline value was -26.6 +/- 3.8 percent and -28.8 +/- 4.3 percent, respectively (both p less than 0.01), but was significantly smaller (-16.1 +/- 3.6 percent and -19.2 +/- 4.1 percent, respectively [p less than 0.05]) when atrial transport was maintained during sequential A-V pacing at the same heart rate. During simultaneous V/A pacing at 160 beats/min, two thirds of the reduction in end-diastolic and stroke volumes from the baseline value was due to the increment in heart rate as assessed from sequential A-V pacing and the other third was due to loss of atrial transport. The data indicate that the hemodynamic consequences of supraventricular tachyarrhythmias in patients with normal ventricular function are due primarily to decreases in ventricular volume as heart rate is increased and atrial contribution is lost rather than to any changes in left ventricular ejection fraction.


Repurposing of Copper(II)-chelating Drugs for the Treatment of Neurodegenerative Diseases.
There is mounting urgency to find new drugs for the treatment of neurodegenerative disorders. A large number of reviews have exhaustively described either the molecular or clinical aspects of neurodegenerative diseases such as Alzheimer's (AD) and Parkinson's (PD). Conversely, reports outlining how known drugs in use for other diseases can also be effective as therapeutic agents in neurodegenerative diseases are less reported. This review focuses on the current uses of some copper(II) chelating molecules as potential drug candidates in neurodegeneration. Starting from the well-known harmful relationships existing between the dyshomeostasis and mis-management of metals and AD onset, we surveyed the experimental work reported in the literature, which deals with the repositioning of metal-chelating drugs in the field of neurodegenerative diseases. The reviewed papers were retrieved from common literature and their selection was limited to those describing the biomolecular aspects associated with neuroprotection. In particular, we emphasized the copper(II) coordination abilities of the selected drugs. Copper, together with zinc and iron, are known to play a key role in regulating neuronal functions. Changes in copper homeostasis are crucial for several neurodegenerative disorders. The studies included in this review may provide an overview on the current strategies aimed at repurposing copper (II) chelating drugs for the treatment of neurodegenerative disorders. Starting from the exemplary case of clioquinol repurposing, we discuss the challenge and the opportunities that repurposing of other metal-chelating drugs may provide (e.g. PBT-2, metformin and cyclodipeptides) in the treatment of neurodegenerative disease. In order to improve the success rate of drug repositioning, comprehensive studies on the molecular mechanism and therapeutic efficacy are still required. The present review upholds that drug repurposing makes significant advantages over drug discovery since repositioned drugs had already passed the safety and toxicity tests. Promising drug candidates in neurodegenerative diseases may be represented by copper chelating classes of drugs, provided that sufficient details on their mechanism of action are available to encourage further investigations and clinical trials.


Evidence of asymptomatic submicroscopic malaria in low transmission areas in Belaga district, Kapit division, Sarawak, Malaysia.
Malaysia has declared its aim to eliminate malaria with a goal of achieving zero local transmission by the year 2020. However, targeting the human reservoir of infection, including those with asymptomatic infection is required to achieve malaria elimination. Diagnosing asymptomatic malaria is not as straightforward due to the obvious lack of clinical manifestations and often subpatent level of parasites. Accurate diagnosis of malaria is important for providing realistic estimates of malaria burden and preventing misinformed interventions. Low levels of parasitaemia acts as silent reservoir of transmission thus remains infectious to susceptible mosquito vectors. Hence, the aim of this study is to investigate the prevalence of asymptomatic submicroscopic malaria (SMM) in the District of Belaga, Sarawak. In 2013, a total of 1744 dried blood spots (DBS) were obtained from residents of 8 longhouses who appeared healthy. Subsequently, 251 venous blood samples were collected from residents of 2 localities in 2014 based on the highest number of submicroscopic cases from prior findings. Thin and thick blood films were prepared from blood obtained from all participants in this study. Microscopic examination were carried out on all samples and a nested and nested multiplex PCR were performed on samples collected in 2013 and 2014 respectively. No malaria parasites were detected in all the Giemsa-stained blood films. However, of the 1744 samples, 29 (1.7%) were positive for Plasmodium vivax by PCR. Additionally, of the 251 samples, the most prevalent mono-infection detected by PCR was Plasmodium falciparum 50 (20%), followed by P. vivax 39 (16%), P. knowlesi 9 (4%), and mixed infections 20 (8%). This research findings conclude evidence of Plasmodium by PCR, among samples previously undetectable by routine blood film microscopic examination, in local ethnic minority who are clinically healthy. SMM in Belaga district is attributed not only to P. vivax, but also to P. falciparum and P. knowlesi. In complementing efforts of programme managers, there is a need to increase surveillance for SMM nationwide to estimate the degree of SMM that warrant measures to block new transmission of malaria.


Direct radioactive labelling of poly(ADP-ribose) in developing Xenopus laevis embryos.
It has not previously been possible to label the nuclear protein modification poly(ADP-ribose) directly from NAD because of the impermeability of the cell membrane. We have overcome this important problem by micro-injection of radioactively labelled NAD into Xenopus laevis early embryos. The polymer was identified and then quantified by its insensitivity to DNAase, RNAase, and spleen phosphodiesterase and by the chromatographic mobility of the products of digestion with snake-venom phosphodiesterase. The quantity of poly(ADP-ribose) present after 25 h of development (129 ng/mg DNA) is lower than that found in fully differentiated tissue.


Identification of a novel proapoptotic function of resveratrol in fat cells: SIRT1-independent sensitization to TRAIL-induced apoptosis.
The phytochemical resveratrol has recently gained attention for its protection against metabolic disease and for extension of life span, which have been linked to its metabolic effects and SIRT1 activation in fat cells. However, little is known about the effect of resveratrol on fat cell apoptosis. Here, we identify a novel, SIRT1-independent mechanism by which resveratrol regulates fat cell numbers. We demonstrate for the first time that resveratrol enhances TNF-related apoptosis-inducing ligand (TRAIL)- or CD95-induced apoptosis of human preadipocytes in a highly synergistic manner (EC(50) at 72 h: resveratrol, >300 microM; TRAIL, >100 ng/ml; combination: 30 microM resveratrol and 10 ng/ml TRAIL, combination index 0.4). Similar results in primary human preadipocytes prepared from subcutaneous white adipose tissue and mature human adipocytes underline the relevance to human physiology. Mechanistic studies reveal that resveratrol inhibits PI3K-driven phosphorylation of Akt, leading to increased Bax activation, loss of mitochondrial membrane potential, cytochrome c release, and caspase-dependent apoptosis. The synergistic interaction of resveratrol and TRAIL depends on the intrinsic apoptosis pathway and caspases, since Bcl-2 overexpression and the caspase inhibitor zVAD.fmk inhibit apoptosis, whereas knockdown of SIRT1 by RNA interference has no effect. The discovery of this novel activity of resveratrol significantly advances the knowledge of fat tissue regulation by resveratrol and has important implications for its use in metabolic and age-related diseases.


Necrotizing Soft Tissue Fasciitis after Intramuscular Injection.
Necrotizing soft tissue fasciitis (NSTIs) or necrotizing fasciitis is an infrequent and serious infection. Herein, we describe the clinical course of a female patient who received a diagnosis of NSTIs after gluteus intramuscular injection. We also report the results of our review of published papers from 1997 to 2017. Since now, 19 cases of NSTIs following intramuscular injections have been described. We focus on the correlation between intramuscular injection and NSTIs onset, especially in immunosuppressed patients treated with corticosteroids, suffering from chronic diseases or drug addicted. Intramuscular injections can provoke severe tissue trauma, representing local portal of infection, even if correctly administrated. Otherwise, it is important not to inject drug in subcutaneous, which is a less vascularized area and therefore more susceptible to infections. Likewise, a proper injecting technique and aspiration prior to injection seem to be valid measure to prevent intra-arterial or para-arterial drug injection with the consequent massive inflammatory reaction. Necrosis at the infection site appears to be independent of the drug, and it is a strong additional risk factor for NSTIs.


CD14+CD16++ cells derived in vitro from peripheral blood monocytes exhibit phenotypic and functional dendritic cell-like characteristics.
We previously reported an increased percentage of CD14+CD16++ monocytes in the peripheral blood of HIV-infected patients but the physiopathological role of this monocyte subset remains unclear. Cells with a CD14+CD16++ phenotype may be obtained in vitro by culturing human peripheral blood monocytes in the presence of GM-CSF, IL-4 and IL-10. In the present study, we compared the phenotypic and functional characteristics of monocytes-derived CD14+CD16++ cells with those of macrophages and dendritic cells. We show that the CD14+CD16++ cells express dendritic cell markers: CD40, CD80, CD86, HLA-DR, CD11b, CD11c, CD18, CD1a, and CD83. Using RNase protection assay, we demonstrate that CD14+CD16++ cell subset expresses a low ratio of IL-1beta/IL-1ra mRNA and expresses IL-6, MIP-1alpha, MIP-1beta, MCP-1, IL-8, RANTES and I-309 transcripts, similar to dendritic cells. CD14+CD16++ cells produce IL-12, MCP-1 and IL-8, as assessed by flow cytometry. Moreover, CD14+CD16++ cells pulsed with different recall antigens induce a potent autologous T cell proliferation. Altogether, these results provide evidence that CD14+CD16++ cells differentiated in vitro from peripheral blood monocytes exhibit dendritic cell characteristics.


Safety of radial coronary angiography with uninterrupted direct-acting oral anticoagulant treatment.
It is not known whether direct-acting oral anticoagulants (DOACs), such as dabigatran, apixaban, and rivaroxaban increase the risk of bleeding complications during or after coronary catheterization. The aim of this study was to investigate the safety of uninterrupted DOAC treatment during diagnostic radial coronary angiography (CAG). This study included 160 patients who underwent diagnostic radial cardiac catheterization. The 60 patients in the group who were using a DOAC (apixaban, rivaroxaban, or dabigatran) were enrolled in a Group A. Post-procedure results from patients in Group A were compared with those of an age- and sex-matched control group (Group B) that included 100 patients who underwent radial CAG who did not use a DOAC. There was no significant difference in the procedure and compression times, creatinine level, or presence of hypertension, diabetes mellitus, smoking, alcohol use, vascular disease, or congestive heart failure between the 2 groups. During the 1 -month follow-up period, only 1 radial occlusion was registered in the control group (Group B). There was no case of a large hematoma (>5 cm or extending to the forearm), dissection, fistula, perforation, or compartment syndrome. Hematomas smaller than 5 cm were seen in 2 patients (1 in each group). No thrombotic events were observed during follow-up examinations. Performing radial CAG with uninterrupted DOAC treatment appears to carry no risk of increased early or short-term complications. The simple, uninterrupted DOAC strategy is comfortable, easy, and safe.


Microbeam radiation therapy - grid therapy and beyond: a clinical perspective.
Microbeam irradiation is spatially fractionated radiation on a micrometer scale. Microbeam irradiation with therapeutic intent has become known as microbeam radiation therapy (MRT). The basic concept of MRT was developed in the 1980s, but it has not yet been tested in any human clinical trial, even though there is now a large number of animal studies demonstrating its marked therapeutic potential with an exceptional normal tissue sparing effect. Furthermore, MRT is conceptually similar to macroscopic grid based radiation therapy which has been used in clinical practice for decades. In this review, the potential clinical applications of MRT are analysed for both malignant and non-malignant diseases.


Balneotherapy in fibromyalgia: a single blind randomized controlled clinical study.
We aimed to evaluate the effectiveness of balneotherapy in fibromyalgia management. Fifty women with fibromyalgia under pharmacological treatment were randomly assigned to either the balneotherapy (25) or the control (25) group. Four patients from the balneotherapy group and one patient from the control group left the study after randomization. The patients in the balneotherapy group (21) had 2 thermomineral water baths daily for 2 weeks in Tuzla Spa Center. The patients in the control group (24) continued to have their medical treatment and routine daily life. An investigator who was blinded to the study arms assessed the patients. All patients were assessed four times; at the beginning of the study, at the end of the 2nd week, the 1st month, and the 3rd month after balneotherapy. Outcome measures of the study were pain intensity, Fibromyalgia Impact Questionnaire (FIQ), Beck Depression Inventory (BDI), patient's global assessment, investigator's global assessment, SF-36 scores, and tender point count. Balneotherapy was found to be superior at the end of the cure period in terms of pain intensity, FIQ, Beck Depression Inventory, patient's global assessment, investigator's global assessment scores, and tender point count as compared to the control group. The superiority of balneotherapy lasted up to the end of the 3rd month, except for the Beck Depression Inventory score and the investigator's global assessment score. Significant improvements were observed in PF, GH, and MH subscales of SF-36 during the study period in the balneotherapy group; however, no such improvement was observed in the control group. Balneotherapy was superior only in VT subscale at the end of therapy and at the end of the third month after the therapy as compared to the controls. It was concluded that balneotherapy provides beneficial effects in patients with fibromyalgia.


Viability of Iberian x Meishan F2 newborn pigs. I. Analysis of physiological and vitality variables.
Eight physiological and vitality variables related to the first minutes of the life of Iberian x Meishan F2 piglets were analyzed. Variables included rectal temperature, arterial oxygen saturation, heart rate (all were monitored at birth and 60 min later), time to reach the udder, and time to the first colostrum intake. Litter effect within boar, a random source of variation, influenced all variables, whereas the random boar effect was not significant for heart rate and arterial oxygen saturation at birth and the time to reach the udder. Birth weight influenced rectal temperature at birth (P < 0.01) and 60 min later (P < 0.001), arterial oxygen saturation 1 h after birth (P < 0.05), heart rate 1 h after birth (P < 0.001), time to reach the udder (P < 0.05), and time to the first suckle (P < 0.01), whereas viability score influenced rectal temperatures (at birth and 60 min later; P < 0.001), and time to reach the udder (P < 0.001) and to suckle (P < 0.001). Finally, the order of birth showed significant effects for rectal temperature 1 h after birth (P < 0.001) and time to first colostrum intake (P < 0.001). Correlation coefficients between physiological variables were generally low, with the exception of the ones for rectal temperature 1 h after birth and arterial oxygen saturation 1 h after birth (0.38; P < 0.001), and for rectal temperature at birth and rectal temperature 1 h after birth (0.34; P < 0.001). Times to reach the udder and to suckle were highly correlated (0.67; P < 0.001) and also were moderately and negatively correlated with rectal temperature 1 h after birth (-0.36 and -0.38 respectively; P < 0.001). Heart rates at birth and 1 h later, as well as arterial oxygen saturation at birth and 1 h later, were not correlated, showing that the values at birth do not necessarily provide information about the physiological status of the pig 1 h after birth. The recording of physiological variables, birth weight, and viability score may be useful to identify weak piglets quickly and to establish palliative measures.


Xenopus oocyte maturation does not require new cyclin synthesis.
Progesterone induces fully grown, stage VI, Xenopus oocytes to pass through meiosis I and arrest in metaphase of meiosis II. Protein synthesis is required twice in this process: in order to activate maturation promoting factor (MPF) which induces meiosis I, and then again after the completion of meiosis I to reactivate MPF in order to induce meiosis II. We have used antisense oligonucleotides to destroy maternal stores of cyclin mRNAs, and demonstrate that new cyclin synthesis is not required for entry into either meiosis I or II. This finding is consistent with the demonstration that stage VI oocytes contain a store of B-type cyclin polypeptides (Kobayashi, H., J. Minshull, C. Ford, R. Golsteyn, R. Poon, and T. Hunt. 1991. J. Cell Biol. 114:755-765). Although approximately 70% of cyclin B2 is destroyed at first meiosis, the surviving fraction, together with a larger pool of surviving cyclin B1, must be sufficient to allow the reactivation of MPF and induce entry into second meiotic metaphase. Since stage VI oocytes do not contain any cyclin A, our results show that cyclin A is not required for meiosis in Xenopus. We discuss the possible nature of the proteins whose synthesis is required to induce meiosis I and II.


Immunohistochemical study of epithelial-myofibroblast interaction in Barrett metaplasia.
Sub-epithelial myofibroblasts are known to influence the biology (proliferation, differentiation and apoptosis) of overlying epithelia. In the intestine, myofibroblasts have been demonstrated to be essential for epithelial differentiation. It is therefore hypothesized that myofibroblasts may also be involved in intestinal metaplasia that is characteristic of Barrett esophagus. This study endeavors to immunohistologically evaluate epithelial-myofibroblast interaction in Barrett's metaplasia. Nineteen archival esophageal endoscopic biopsies of Barrett's metaplasia were immune-phenotyped for the following epithelial and myofibroblast antigens - cytokeratins (CK) 8, 13, 18, CDX2 (Caudal type homeobox 2), a-smooth muscle actin (SMA). alpha-SMA immunostaining revealed close association between myofibroblasts and metaplastic Barrett's epithelium but not with normal esophageal squamous epithelium. Myofibroblasts were more prominent in dysplastic than in non-dysplastic Barrett metaplasia. CDX2 and CK 8/18, indicators of intestinal differentiation were expressed in Barrett metaplasia but not normal esophageal squamous epithelium, while the reverse was the case for CK 13, which only stained normal esophageal squamous epithelium. Although their precise role is yet to be clearly defined, sub-epithelial myofibroblasts are very likely involved in the pathogenesis of Barrett's metaplasia.


Chromosomal gains measured in cytology samples from women with abnormal cervical cancer screening results.
Chromosomal gains at 3q26, 5p15 and 20q13 have been described in cervical precancer and cancer. We evaluated a novel fluorescence in situ hybridization (FISH) assay that detects gains at these three loci simultaneously as a possible biomarker for detecting cervical precancer. Chromosomal copy numbers at 3q26, 5p15, 20q13 and the centromere of chromosome7 (cen7) in liquid-based cytology specimens from 168 women enrolled in the Biopsy Study were determined by FISH. The number of cells with ≥ 3 or ≥ 4 signals for a genomic locus was enumerated and diagnostic test performance measures were calculated using receiver operating characteristic (ROC) analyses. Sensitivity and specificity values were determined for the detection of CIN2+ and/or HSIL. The median number of cells with ≥ 3 signals increased with the severity of cervical lesion for each genomic locus (p-trend<0.02 for each locus). ROC analysis for the number of cells with ≥ 3 signals resulted in area under the curve values of 0.70 (95% CI: 0.54-0.86), 0.67 (0.52-0.83), 0.67 (0.51-0.83) and 0.78 (0.64-0.92) for 3q26, 5p15, 20q13 and cen7, respectively, for the detection of CIN2+ and/or HSIL. Positivity for gains at multiple loci resulted in only slightly better test performance measures than those for the individual probes for four distinct combinations of probes. Chromosomal gains at 3q26, 5p15, 20q13 and cen7 are associated with severity of cervical lesions. Further studies are required to quantify risk stratification of FISH assays for cervical cancer screening.


The absence of protein Y4yS affects negatively the abundance of T3SS Mesorhizobium loti secretin, RhcC2, in bacterial membranes.
Mesorhizobium loti MAFF303099 has a functional type III secretion system (T3SS) that is involved in the determination of nodulation competitiveness on Lotus. The M. loti T3SS cluster contains gene y4yS (mlr8765) that codes for a protein of unknown function (Y4yS). A mutation in the y4yS gene favors the M. loti symbiotic competitive ability on Lotus tenuis cv. Esmeralda and affects negatively the secretion of proteins through T3SS. Here we localize Y4yS in the bacterial membrane using a translational reporter peptide fusion. In silico analysis indicated that this protein presents a tetratricopeptide repeat (TPR) domain, a signal peptide and a canonical lipobox LGCC in the N-terminal sequence. These features that are shared with proteins required for the formation of the secretin complex in type IV secretion systems and in the Tad system, together with its localization, suggest that the y4yS-encoded protein is required for the formation of the M. loti T3SS secretin (RhcC2) complex. Remarkably, analysis of RhcC2 in the wild-type and M. loti y4yS mutant strains indicated that the absence of Y4yS affects negatively the accumulation of normal levels of RhcC2 in the membrane.


Effects of rosuvastatin on the production and activation of matrix metalloproteinase-2 and migration of cultured rat vascular smooth muscle cells induced by homocysteine.
To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) and on cell migration of cultured rat vascular smooth muscle cells (VSMCs). Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Rat VSMCs were incubated with different concentrations of homocysteine (50-5000 μmol/L). Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin (10(-9)-10(-5) mol/L) were added when VSMCs were induced with 1000 μmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Homocysteine (50-1000 μmol/L) increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5000 μmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and activation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine (50-5000 μmol/L) stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Homocysteine (50-1000 μmol/L) significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and activation of MMP-2 and abnormal migration of VSMCs induced by homocysteine.


[Toxicological aspects of the use of hormones as anabolic agents (author's transl)].
The use of anabolic agents may give rise to residues in foods of animal origin and thus to potential effects on public health. Therefore, the physiology, metabolism and toxicity of the three endogenous hormones. 17 beta-oestradiol, progesterone and testosterone are discussed. In addition, the effects of the use of three exogenous hormones, diethylstilboestrol (DES), trenbolone and zeranol are evaluated. As can be expected all six compounds have effects on the endocrine system and reproduction. Endogenous hormones and DES are teratogenic at high-dose levels. With the exception of DES, there is no evidence of a mutagenic action. 17 beta-Oestradiol and testosterone were shown to induce tumours at high-dose levels in experimental animals; probably however, a threshold can be established. The residues in edible tissue resulting from the use of endogenous hormones are much smaller than are the natural levels in cow's milk and butter. In view of the carcinogenicity of DES in man, its use should be strongly discouraged. A definite statement on the acceptability of the use of trenbolone and zeranol cannot be made.


Occurrence of microplastics in raw and treated drinking water.
The study investigates the content of microplastic particles in freshwater and drinking water. Specifically, three water treatment plants (WTPs) supplied by different kinds of water bodies were selected and their raw and treated water was analysed for microplastics (MPs). Microplastics were found in all water samples and their average abundance ranged from 1473 ± 34 to 3605 ± 497 particles L-1 in raw water and from 338 ± 76 to 628 ± 28 particles L-1 in treated water, depending on the WTP. This study is one of very few that determine microplastics down to the size of 1 μm, while MPs smaller than 10 μm were the most plentiful in both raw and treated water samples, accounting for up to 95%. Further, MPs were divided into three categories according to their shape. Fragments clearly prevailed at two of the WTPs and fibres together with fragments predominated at one case. Despite 12 different materials forming the microplastics being identified, the majority of the MPs (>70%) comprised of PET (polyethylene terephthalate), PP (polypropylene) and PE (polyethylene). This study contributes to fill the knowledge gap in the field of emerging microplastic pollution of drinking water and water sources, which is of concern due to the potential exposure of microplastics to humans.


FACE facts: why human genetics will always provoke bioethics.
Some people dispute the relative importance of issues in genetics and biotechnology for the future of bioethics, either because they think the problems are time-limited or because they give priority to issue of human rights and social justice in health care. In fact, the special historical standing of genetic issue s in bioethics reflects four overlapping sources of moral sensitivity which ar inherent in the stories that genetic science tells and raise paradigmatic justice concerns: the implications of new genetic knowledge for people's understanding of their familial roles, ancestral origins, community memberships, and ethnic affiliations. Beneath worries over "genetic privacy," "the therapeutic gap," and the "post-human," this constellation of basic wellspring which both insures and justifies a central place for genetics on the agenda of bioethics.


[Comparison of the verapamil concentration of human blood serum and saliva].
Verapamil concentrations were measured in the samples of blood serum and saliva taken at the same time in patients with angina of effort within 0.5-24 hr after administration of a single dose of the drug (14 persons) or the last dose of the course (10 persons). A mean ratio of saliva verapamil concentration to blood serum concentration was 0.85 +/- 0.09 (from 0.19 to 2.69) after a single administration and 0.77 +/- 0.04 (from 0.29 to 1.09) after the course treatment. A statistically significant (p less than 0.001) positive correlation was revealed between verapamil concentrations in blood serum and saliva after a single administration (r = 0.69) and course treatment (r = 0.80). It is suggested that determination of verapamil concentration in the saliva of patients on long-term verapamil medication should be used for a reference estimation of its level in the blood.


Mediastinal lymphadenopathy in patients with or without previous malignancy: EUS-FNA-based differential cytodiagnosis in 153 patients.
Mediastinal lymphadenopathy (ML) is a cause for concern, especially in patients with previous malignancy. The investigation of choice is thoracic CT with a variable sensitivity and specificity requiring tissue diagnosis. We used endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) for cytodiagnosis of ML in patients with and without previous malignancy. The cause, distribution of lesions, and incidence of second cancers were investigated. Linear echoendoscopes and 22-gauge needles for cytology were used for EUS-FNA. A cytological diagnosis of malignancy was accepted, and histology or consistent follow-up of at least 9 months confirmed benign results. One hundred fifty-three patients underwent EUS-FNA between November 1997 and November 1999 (mean age, 60 yr; range, 13-82 yr; 105 men). Cytology was adequate in 150 patients. Final diagnosis was malignancy in 84 and benign in 66 patients (sensitivity, specificity, and diagnostic accuracy: 92%, 100%, 95%, respectively). In 101 patients without previous cancer cytology identified 48 malignant (lung, 41; extrathoracic, 7) and 51 benign lesions (inflammation, 35; various, 9; sarcoidosis, 7) (sensitivity, specificity, accuracy: 88%, 100%, 94%). Fifty-two patients had prior malignancy, mostly in extrathoracic sites. Cytology revealed recurrences in 21 patients, second cancer in 9 and benign lesions in 21 patients (inflammatory, 11; sarcoidosis, 8; tuberculosis, 1; abscess, 1) (sensitivity, specificity, accuracy: 97%, 100%, 98%). In patients without previous cancer malignant ML originates from the lung >80%. In those with previous malignancy recurrence of extrathoracic sites is the major cause. Benign lesions and treatable second cancers occur in a significant frequency, emphasizing the need for tissue diagnosis. EUS-FNA is a safe and minimally invasive alternative for cytodiagnosis in the mediastinum.


Endogenous IL-17 as a mediator of neutrophil recruitment caused by endotoxin exposure in mouse airways.
We have previously demonstrated that administration of the recently described cytokine IL-17 in rat airways in vivo recruits and activates neutrophils locally. In the current study, we examined whether endogenous IL-17 is involved in mediating neutrophil recruitment caused by endotoxin exposure in mouse airways. Our in vivo data show that local endotoxin exposure causes the release of free, soluble IL-17 protein 6 h later. Systemic pretreatment with a neutralizing anti-IL-17 Ab almost completely inhibits neutrophil recruitment 24 h, but not 6 h, after endotoxin exposure in the airways. Pretreatment with neutralizing anti-IL-6 and anti-macrophage inflammatory protein (MIP)-2 Abs inhibits neutrophil recruitment caused by local endotoxin exposure and IL-17, respectively. Our in vitro data show that endotoxin exposure stimulates the release of soluble IL-17 protein in T lymphocytes harvested from lung and spleen, respectively, and that this cytokine release requires coculture with airway macrophages. Intracellular IL-17 protein is detected in T lymphocytes from spleen but not in airway macrophages after coculture and stimulation of these two cell types. Finally, anti-IL-17 does not alter endotoxin-induced release of IL-6 and MIP-2 from T lymphocytes and airway macrophages in coculture. In conclusion, our results indicate that endotoxin exposure causes the release of IL-17 from T lymphocytes and that this cytokine release requires the presence of macrophages. Once released, endogenous IL-17 acts in part by inducing local release of neutrophil-mobilizing cytokines such as IL-6 and MIP-2, from nonlymphocyte, nonmacrophage cells, and this contributes to recruitment of neutrophils in the airways. These IL-17-related mechanisms constitute potential targets for pharmacotherapy against exaggerated neutrophil recruitment in airway disease.


Whole blood preservation methods alter chemokine receptor detection in mass cytometry experiments.
A potential hurdle when applying mass cytometry to the field study setting is the streamlining of sample collection while at the same time protecting the integrity of important cell epitopes. Whole blood preservation kits applying fixation and/or permeabilization agents are increasingly used in clinical trials to preserve leukocytes needed for downstream analysis. We here present a structured overview of leukocyte surface marker detectability in samples processed with four commercially available whole blood preservation kits; 1) Proteomic Stabilizer, 2) Stable-Lyse V2 and Stable-Store V2, 3) Cytodelics and 4) Lyse/Fix Buffer, as well as in samples treated with buffers included in Mass-tag Cellular Barcoding kits. Isolated leukocytes were stained with a 28-marker panel (including 7 chemokine receptors) of metal-conjugated antibodies and analysed on a mass cytometer. Exploration of the data by manual gating and viSNE analysis showed that although many markers were similarly detected across all sample conditions, most of the chemokine receptors in our panel, particularly CXCR3, CCR4, CCR6 and CXCR5, were incorrectly detected in the preserved samples and thus incompatible with the fixation and permeabilization agents found in whole blood preservation kits and in buffers used prior to barcoding.


Strategies for monitoring the emerging polar organic contaminants in water with emphasis on integrative passive sampling.
Although polar organic contaminants (POCs) such as pharmaceuticals are considered as some of today's most emerging contaminants few of them are regulated or included in on-going monitoring programs. However, the growing concern among the public and researchers together with the new legislature within the European Union, the registration, evaluation and authorisation of chemicals (REACH) system will increase the future need of simple, low cost strategies for monitoring and risk assessment of POCs in aquatic environments. In this article, we overview the advantages and shortcomings of traditional and novel sampling techniques available for monitoring the emerging POCs in water. The benefits and drawbacks of using active and biological sampling were discussed and the principles of organic passive samplers (PS) presented. A detailed overview of type of polar organic PS available, and their classes of target compounds and field of applications were given, and the considerations involved in using them such as environmental effects and quality control were discussed. The usefulness of biological sampling of POCs in water was found to be limited. Polar organic PS was considered to be the only available, but nevertheless, an efficient alternative to active water sampling due to its simplicity, low cost, no need of power supply or maintenance, and the ability of collecting time-integrative samples with one sample collection. However, the polar organic PS need to be further developed before they can be used as standard in water quality monitoring programs.


Connexin implication in the control of the murine beta-cell mass.
Diabetes develops when the insulin needs of peripheral cells exceed the availability or action of the hormone. This situation results from the death of most beta-cells in type 1 diabetes, and from an inability of the beta-cell mass to adapt to increasing insulin needs in type 2 and gestational diabetes. We analyzed several lines of transgenic mice and showed that connexins (Cxs), the transmembrane proteins that form gap junctions, are implicated in the modulation of the beta-cell mass. Specifically, we found that the native Cx36 does not alter islet size or insulin content, whereas the Cx43 isoform increases both parameters, and Cx32 has a similar effect only when combined with GH. These findings open interesting perspectives for the in vitro and in vivo regulation of the beta-cell mass.


Interleukin-28b CC genotype predicts early treatment response and CT/TT genotypes predicts non-response in patients infected with HCV genotype 3.
Response to antiviral therapy for hepatitis C virus (HCV) depends upon the genotype and host immune response. IL28b gene mutations have been shown to modulate host antiviral immune response against genotype 1. However, the predictive value of IL28b polymorphism in genotype 3 HCV patients is largely unknown. The association of IL28b polymorphism with virological response was studied in 356 patients with genotype 3 chronic HCV undergoing treatment with peg-interferon and ribavirin and was compared with matched controls. IL28b genotyping followed by DNA sequencing was performed to identify the CC, CT, or TT genotypes. Two log reduction of HCV RNA at Day 7 (Quick Viral Response, QVR) and HCV RNA negativity at Day 28 (Rapid Viral Response, RVR) were analyzed with CC and non-CC genotypes in addition to other predictors of response. The associations of alleles with the response patterns were predicted. Sustained viral response was seen in 250 (70.2%) patients and the IL28b genotype CC/CT/TT distribution was 61.1%; 30.5%; and 8.4%, respectively. The non-CC genotypes were significantly higher in non-responders when compared to responders (67.6% vs. 38.9%, P < 0.001). Interestingly, the rapid viral response in responders was observed in 72.7% with the CC genotype and in 27.2% with the non-CC genotype (P < 0.001). Multivariate analysis showed CC genotype as an independent factor predicting the sustained viral response in patients infected with HCV genotype 3. In conclusion, the IL28b CT/TT genotype strongly correlates with treatment non-response in patients infected with HCV genotype 3 and CC genotype of IL28b is associated with higher quick viral response.


Dianion diagnostics in DESIREE: High-sensitivity detection of Cn2- from a sputter ion source.
A sputter ion source with a solid graphite target has been used to produce dianions with a focus on carbon cluster dianions, Cn2-, with n = 7-24. Singly and doubly charged anions from the source were accelerated together to kinetic energies of 10 keV per atomic unit of charge and injected into one of the cryogenic (13 K) ion-beam storage rings of the Double ElectroStatic Ion Ring Experiment facility at Stockholm University. Spontaneous decay of internally hot Cn2- dianions injected into the ring yielded Cn- anions with kinetic energies of 20 keV, which were counted with a microchannel plate detector. Mass spectra produced by scanning the magnetic field of a 90° analyzing magnet on the ion injection line reflect the production of internally hot C72- - C242- dianions with lifetimes in the range of tens of microseconds to milliseconds. In spite of the high sensitivity of this method, no conclusive evidence of C62- was found while there was a clear C72- signal with the expected isotopic distribution. This is consistent with earlier experimental studies and with theoretical predictions. An upper limit is deduced for a C62- signal that is two orders-of-magnitude smaller than that for C72-. In addition, CnO2- and CnCu2- dianions were detected.


Accuracy of Remote Hearing Assessment in a Rural Community.
This study determined the accuracy of pure tone air conduction (AC) thresholds obtained using a synchronous telemedicine approach without a sound booth in a rural South African community. The global need for increased hearing healthcare currently far exceeds the capacity for delivering these services, especially in developing countries. A tele-audiology approach using a portable diagnostic audiometer could provide the solution, enabling hearing assessments to be conducted remotely and without a sound booth. Hearing thresholds in a sound booth and natural environment were obtained from an initial sample of 20 adults (age range, 19-63 years; mean age, 50±13 years; 55% female), recruited from a rural agricultural community. A subgroup of 10 adults (20 ears) volunteered for the telemedicine threshold testing. AC thresholds (250-8,000 Hz) were determined and subsequently compared in these environments. Typical threshold variability was determined using test-retest correspondence as a reference for the threshold correspondence using a telemedicine mode. Test-retest threshold correspondence in the booth and natural environments was within±5 dB in 96.7% and 97.5% of comparisons, respectively. No significant differences were obtained in AC hearing thresholds determined in the telemedicine configuration compared with those recorded in the gold standard booth environment. Threshold correspondence between the telemedicine compared with booth and natural environments were within±5 dB in 82% and 85% of comparisons, respectively. The current study demonstrates the validity of using synchronous telemedicine for conducting hearing assessments in a remote rural agricultural community without a sound booth.


Mental health care consumers' relative valuing of clinician performance information.
Research demonstrates significant variability in mental health clinicians' overall and domain-specific effectiveness with their patients. Despite calls to increase patient access to performance information, little is known about patients' relative valuing of this information in the context of other treatment factors. We aimed to obtain an understanding of patients' relative valuing of provider performance track records and other therapist and treatment characteristics for their mental health care decision making. Participants were 403 (Mage = 41.20; 66.5% female) community mental health patients who completed a multicomponent survey that included an adapted delayed-discounting paradigm to examine the relative valuing. Multiple descriptive, quantitative indices of relative valuing were calculated, as well as an exploratory latent profile analysis to ascertain the presence of homogenous relative-valuing subgroups. Overall, participants valued provider track record information. They also evidenced relatively higher preference values for working with therapists who had specific efficacy in treating their primary problem domain, charged less, and with whom there is a high likelihood of establishing a good alliance. Two latent profiles were identified: one representing higher valuing of provider performance and another consistently representing less emphasis. Participants with higher track-record valuing were younger, believed that therapists are not interchangeable, and endorsed trust in the collection and use of performance information. Harnessing clinician information to make more personalized and informed treatment decisions could potentially promote better treatment engagement, retention, and outcomes. (PsycINFO Database Record


Human immunodeficiency virus-related oral manifestations and gender. A longitudinal analysis. The University of California, San Francisco Oral AIDS Center Epidemiology Collaborative Group.
Because human immunodeficiency virus (HIV) infection affects an increasing number of women in the United States, we investigated the role played by gender in the occurrence of HIV-related oral conditions. As part of a 4-year prospective study of 3 epidemiological cohorts, oral and physical examinations (including blood tests) were performed on HIV-infected men (n = 200) and women (n = 218) at 6-month intervals. Our outcome variables included oral conditions commonly associated with HIV infection: hairy leukoplakia, candidiasis, ulcers, warts, non-Hodgkin lymphoma, Kaposi sarcoma, and parotid enlargement. Only hairy leukoplakia, candidiasis, and ulcers were observed. The occurrence of hairy leukoplakia and candidiasis was higher in men (22% and 24%, respectively) than in women (9% and 13%, respectively) during the study period. A regression model for longitudinal data (generalized estimating equation) disclosed that the odds of having hairy leukoplakia were 2.5 times higher for men than for women, after controlling for CD4+ cell count, race, and injecting drug use (95% confidence interval, 1.34-4.76; P = .003). Length of follow-up did not confound this association. A weaker association was found between the occurrence of oral candidiasis and gender (adjusted odds ratio, 1.85; 95% confidence interval, 1.0-3.43; P = .05). In this sample of HIV-infected adults, we found that men were significantly more likely to have hairy leukoplakia than were women. The hairy leukoplakia-gender association merits further investigation, because it may be related to a gender difference in the mode of expression of Epstein-Barr virus.


Study of the association of 17 lipid-related gene polymorphisms with coronary heart disease.
Blood lipids are well-known risk factors for coronary heart disease (CHD). The aim of this study was to explore the association between 17 lipid-related gene polymorphisms and CHD. The current study examined with 784 CHD cases and 739 non-CHD controls. Genotyping was performed on the MassARRAY iPLEX® assay platform. Our analyses revealed a significant association of APOE rs7259620 with CHD (genotype: χ2=6.353, df=2, p=0.042; allele: χ2=5.05, df=1, p=0.025; recessive model: χ2=5.57, df=1, p=0.018). A further gender-based subgroup analysis revealed significant associations of APOE rs7259620 and PPAP2B rs72664392 with CHD in males (genotype: χ2=8.379, df=2, p=0.015; allele: χ2=5.190, df=1, p=0.023; recessive model: χ2=19.3, df=1, p<0.0001) and females (genotype: χ2=9.878, df=2, p=0.007), respectively. Subsequent breakdown analysis by age showed that CETP rs4783961, MLXIPL rs35493868, and PON2 rs12704796 were significantly associated with CHD among individuals younger than 55 years of age (CETP rs4783961: χ2=8.966, df=1, p=0.011 by genotype; MLXIPL rs35493868: χ2=4.87, df=1, p=0.027 by allele; χ2=4.88, df=1, p=0.027 by dominant model; PON2 rs12704796: χ2=6.511, df=2, p=0.039 by genotype; χ2=6.210, df=1, p=0.013 by allele; χ2=5.03, df=1, p=0.025 by dominant model). Significant allelic association was observed between LEPR rs656451 and CHD among individuals older than 65 years of age (χ2=4.410, df=1, p=0.036). Our study revealed significant associations of APOE, PPAP2B, CETP, MLXIPL, PON2, and LEPR gene polymorphisms with CHD among the Han Chinese.


Liver regeneration and hepatic microsomal enzyme induction by acetaminophen and derivatives.
Liver regeneration was stimulated in male rats with two-thirds of the liver removed by feeding a basal diet supplemented with acetaminophen (0.35-1.5%; weight basis), 2-acetamidophenol (1.0%) and acetophenetidin (1.0%) over a period of 10 days po, but was in the control range with the m-isomer, 3-acetamidophenol (1.0%), N-butyryl-p-aminophenol (1.0%), o-, m- and p-aminophenols (0.50%) and 4-acetamidothiophenol. In fact, the latter inhibited at a level of 0.60%. The operated young or mature female underwent no significant increase in control response with acetaminophen (1-1.5%). However, as with the male, the wet and dry liver weight percentages were markedly increased in the intact female fed acetaminophen (1.0-1.5%) as also with 2-acetamidophenol (1.0%). Liver enlargement occurred in the intact male with acetophenetidin (1.0%) but not with the N-butyryl- and thiophenol derivatives fed at 1.0 and 0.50%, respectively. Hepatic microsomal preparations from the intact and operated series showed no remarkable changes in cytochrome P-450 nor in the enzymes, aminopyrine demethylase and benzo[a]pyrene hydroxylase, with the more polar acetaminophen and the N-butyryl compound but the enzymes were elevated in the group fed acetophenetidin. Inductive effects on microsomal enzymes were further amplified by injection of several animals per group with phenobarbital ip daily at 80 mg/kg for the last 3 days prior to sacrifice. Increases in increments or liver weight percentages ensued over the basal values and as investigated in an intact male series, the enzymes ranged higher than the uninjected controls and with the thiophenol-fed group, exceeded those of the phenobarbital-injected controls.


Associations between peripheral Plasmodium falciparum malaria parasitemia, human immunodeficiency virus, and concurrent helminthic infection among pregnant women in Malawi.
Approximately 2 billion persons worldwide are infected with schistosomiasis and soil-transmitted helminths (STH), many in areas where endemic malaria transmission coexists. Few data exist on associations between these infections. Nested within a larger clinical trial, primigravid and secundigravid women provided blood samples for human immunodeficiency virus (HIV) testing and peripheral malaria films and stool and urine for evaluation of STH and Schistosoma spp. during their initial antenatal clinic visit. The most common parasitic infections were malaria (37.6%), S. haematobium (32.3%), and hookworm (14.4%); 14.2% of women were HIV-infected. S. haematobium infection was associated with lower malarial parasite densities (344 versus 557 parasites/μL blood; P < 0.05). In multivariate analysis, HIV and hookworm infection were independently associated with malaria infection (adjusted odds ratio = 1.9 and 95% confidence interval = 1.2-3.0 for HIV; adjusted odds ratio = 1.9 and 95% confidence interval = 1.03-3.5 for hookworm). Concurrent helminthic infection had both positive and negative effects on malaria parasitemia among pregnant women in Malawi.


Risk factors for tibial tuberosity fracture after tibial plateau leveling osteotomy in dogs.
To evaluate factors that predispose to tibial tuberosity (TT) fracture after tibial plateau leveling osteotomy (TPLO) in dogs. Retrospective study. Dogs (n=182) with cranial cruciate ligament (CCL) rupture undergoing 213 TPLO surgeries. Medical records and radiographs of 2 groups of dogs that had TPLO surgery (2000-2001, 2004-2005) were evaluated to determine the effect of operative technique and surgeon experience on TT fracture. TT fracture was diagnosed in 8 dogs (9 TPLO, 4.2% of surgical procedures). Four fractures occurred after unilateral TPLO in 167 dogs (2.4%), 4 fractures occurred after simultaneous bilateral TPLO in 5 dogs (40%), and 1 fracture occurred after staged bilateral TPLO in 36 dogs (2.8%). Simultaneous bilateral TPLO resulted in a 12.4 times higher odds of TT fracture versus unilateral TPLO (P=.046). The mean absolute thickness of the TT after TPLO was less in dogs sustaining TT fractures (7.2 +/- 2.2 mm) than those that did not (10.8 +/- 2.7 mm, P<.0001). The odds of fracture decreased by 37% when the absolute TT width postosteotomy increased by 1 mm (P<.0001). An increase in tibial plateau angle at follow-up versus immediately postoperative was associated with TT fracture (P=.025). Surgeon experience was not associated with TT fracture. A combination of surgical decision-making and surgical technique play a role in the occurrence of TT fracture after TPLO. Simultaneous bilateral TPLO was associated with a high percentage of TT fracture. Careful planning of osteotomy positioning is advised while performing TPLO surgery.


Summary Care Records in urgent and emergency care in England.
Summary Care Records (SCRs) have been created for more than 50% of the population of England. The number is increasing at about 100,000 records a week. Fewer than 1.5% of people have elected not to have a SCR. SCRs contain updated details of patient medication, allergies and adverse reactions, electronically extracted from the GP record. A patient and their GP can also agree to have additional information included. SCRs are being viewed by authorised healthcare staff in urgent and emergency care settings all over England. Benefits are being reported in relation to increased patient safety, improved clinical decision making, improved efficiency and improved quality of care. NHS England strongly supports the uptake and adoption of SCRs by Trusts in England.


A mimotope peptide-based dual-signal readout competitive enzyme-linked immunoassay for non-toxic detection of zearalenone.
In this study, a mimotope peptide-based non-toxic photoelectrochemical (PEC) competitive enzyme-linked immunoassay (ELISA) was established for ultrasensitive detection of zearalenone (ZEN) with dual-signal readout. Using the phage display technique (PDT), a mimotope peptide of ZEN could be harvested by selecting a peptide from a phage-display peptide library, which avoided using mycotoxin itself and minimized potential damage to operators. The tyramine-modified rutile TiO2 mesocrystals (Tyr-RMC) with outstanding PEC properties were utilized as reporter units to label the mimotope peptide. The fabricated peptide@Tyr-RMC probe could anchor on the antibody-modified electrode via competitive immune recognition of free target ZEN. When subjected to catalysis of HRP-H2O2, the Tyr-RMC composite was deposited at the enzyme reaction site, causing rolling circle extension of the reporter unit chains. By merit of the brilliant signal amplification effect of tyramine signal amplification (TSA), dramatically enhanced photocurrent response and enormously increased electrochemical impedance could be determined. Combining all of these advantages, the developed dual-signal readout non-toxicity immunoassay could effectively decrease environmental interference. Also, the designed dual-signal readout biosensor demonstrated a wide linear range between 10-6 and 1 ng mL-1 with a low detection limit of 1 × 10-6 ng mL-1, which provides a valuable reference for developing highly efficient, secure and sensitive detection methods and indicates promising applicability in food testing.


Evaluating the effectiveness of enhancing resilience in human service professionals using a retreat-based Mindfulness with Metta Training Program: a randomised control trial.
This paper explores the feasibility of a brief Mindfulness with Metta Training Program (MMTP), targeting the enhancement of mindfulness and self-compassion in a retreat format, as a means of increasing resilience in human services professionals. In this randomised control trial, 44 human services professionals were randomly allocated either to a MMTP retreat group or to a control group. Following the MMTP intervention, no significant differences between the retreat and control groups were found on resilience, mindfulness and self-compassion variables. However, significant improvements were observed over time for the retreat group for mindfulness and self-compassion at one and four months and for resilience at four-months post MMTP intervention. The results of this pilot study show that MMTP in a retreat format is a promising method of increasing resilience, mindfulness and self-compassion in human services professionals.


Drinker Types, Harm, and Policy-Related Variables: Results from the 2011 International Alcohol Control Study in New Zealand.
The aim was to identify a typology of drinkers in New Zealand based on alcohol consumption, beverage choice, and public versus private drinking locations and investigate the relationship between drinker types, harms experienced, and policy-related variables. Model-based cluster analysis of male and female drinkers including volumes of alcohol consumed in the form of beer, wine, spirits, and ready-to-drinks (RTDs) in off- and on-premise settings. Cluster membership was then related to harm measures: alcohol dependence, self-rated health; and to 3 policy-relevant variables: liking for alcohol adverts, price paid for alcohol, and time of purchase. Males and females were analyzed separately. Men fell into 4 and women into 14 clearly discriminated clusters. The male clusters consumed a relatively high proportion of alcohol in the form of beer. Women had a number of small extreme clusters and some consumed mainly spirits-based RTDs, while others drank mainly wine. Those in the higher consuming clusters were more likely to have signs of alcohol dependency, to report lower satisfaction with their health, to like alcohol ads, and to have purchased late at night. Consumption patterns are sufficiently distinctive to identify typologies of male and female alcohol consumers. Women drinkers are more heterogeneous than men. The clusters relate differently to policy-related variables.


Mechanism of increased glucose uptake by skeletal muscle during E coli endotoxin shock in the dog.
Carbohydrate metabolism of skeletal muscle was studied during 2 mg/kg E coli endotoxin shock in dogs. During natural (free-flow) conditions, glucose uptake by the muscle increased markedly during 6 hours of shock. Increased glucose uptake occurred concomitant with muscle ischemia and hypoxia. However, when muscle blood flow was held constant, thereby preventing local muscle ischemia and hypoxia, glucose uptake by the gracilis muscle did not change during shock. These results implicate local muscle ischemia and/or hypoxia as the mediator(s) of the increased muscle glucose uptake during shock. Further studies demonstrated that local muscle hypoxia was the stimulus for increased glucose uptake by skeletal muscle during endotoxin shock, and muscle ischemia per se did not alter muscle glucose uptake. Since approximately 50% of body mass is composed of skeletal muscle, the contribution of this organ system in the hypoglycemia of endotoxin shock in the dog may be substantial.


Age-related trends in short- and long-term survival after acute myocardial infarction: a 20-year population-based perspective (1975-1995).
This study examines age-related differences and temporal trends in hospital and long-term survival after acute myocardial infarction (AMI) over a 2-decade-long (1975 to 1995) experience. A total of 8,070 patients with validated AMI hospitalized in all acute care hospitals in the Worcester, Massachusetts, metropolitan area (1990 census population 437,000) were studied over 10 one-year periods between 1975 and 1995. This population included 1,326 patients aged <55 years (16.4%), 1,768 patients aged 55 to 64 years (21.9%), 2,325 patients aged 65 to 74 years (28.8%), 1,880 patients aged 75 to 84 years (23.3%), and 771 patients aged > or = 85 years (9.6%). Compared with patients <55 years, patients 55 to 64 years were 2.2 times more likely to die during hospitalization for AMI, whereas patients 65 to 74, 75 to 84, and > or = 85 years were at 4.2, 7.8, and 10.2 times greater risk of dying, respectively. Similar age disparities in the risk of dying were seen when controlling for additional prognostic factors. Despite the adverse impact of increasing age on hospital survival after AMI, declining in-hospital death rates were seen in each of the age groups under study, with declining magnitude of these trends with advancing age. Among discharged hospital patients, increasing age was related to a significantly poorer long-term prognosis. Trends toward improving long-term prognosis were seen in patients discharged in the mid-1990s compared with those discharged in the mid- to late 1970s for patients aged <85 years. The present results demonstrate the marked impact of advancing age on survival after AMI. Despite the adverse impact of age on prognosis, encouraging trends in prognosis were observed in all age groups, although to a lesser extent in the oldest elderly patients. These findings emphasize the low death rates in middle-aged patients with AMI and the need for targeted secondary prevention efforts in elderly patients with AMI.


Mechanisms of cell commitment in myeloid cell differentiation.
The hematopoietic developmental hierarchy originates with a rare population of lymphohematopoietic stem cells that are capable of extensive self-renewal as well as the continuous generation of more developmentally restricted progeny. The generation of mature blood cells from these pluripotent hematopoietic stem cells involves the highly regulated progression through successive stages involving commitment to a specific cell lineage, terminal differentiation of lineage-restricted progenitors, and growth arrest. Although the differentiation commitment of stem cells may be intrinsically determined, it is apparent that a wide variety of external and internal stimuli can influence and modulate lineage choice and differentiation. These factors cooperate with cellular transcription factors to activate or repress the expression of genes responsible for lineage choice, diverse mature phenotypes, and cell cycle progression. The extrinsic and genetic mechanisms that orchestrate the differentiation commitment and myeloid lineage restriction of pluripotent stem cells are of fundamental importance in the regulation of hematopoiesis. The elucidation of these mechanisms of normal myeloid differentiation has provided instrumental insights into the biology of leukemia and other hematopoietic disorders.