A financial analysis of revision hip arthroplasty: the economic burden in relation to the national tariff.
Revision arthroplasty of the hip is expensive owing to the increased cost of pre-operative investigations, surgical implants and instrumentation, protracted hospital stay and drugs. We compared the costs of performing this surgery for aseptic loosening, dislocation, deep infection and peri-prosthetic fracture. Clinical, demographic and economic data were obtained for 305 consecutive revision total hip replacements in 286 patients performed at a tertiary referral centre between 1999 and 2008. The mean total costs for revision surgery in aseptic cases (n = 194) were £11 897 (sd 4629), for septic revision (n = 76) £21 937 (sd 10 965), for peri-prosthetic fracture (n = 24) £18 185 (sd 9124), and for dislocation (n = 11) £10 893 (sd 5476). Surgery for deep infection and peri-prosthetic fracture was associated with longer operating times, increased blood loss and an increase in complications compared to revisions for aseptic loosening. Total inpatient stay was also significantly longer on average (p < 0.001). Financial costs vary significantly by indication, which is not reflected in current National Health Service tariffs.


The association between risk perceptions, anxiety, and self-reported changes in tobacco and nicotine product use due to COVID-19 in May-June 2020 in Israel.
Early in the COVID-19 pandemic, reports about a possible protective effect of nicotine on COVID-19 conflicted with messaging by public health organizations about increased risks of COVID-19 due to smoking. The ambiguous information the public received, combined with COVID-19-induced anxiety, may have led to changes in tobacco or other nicotine product use. This study examined changes in use of combustible cigarettes (CCs), nargila (hookah/waterpipe), e-cigarettes, and IQOS and home-smoking behaviors. We also assessed COVID-19 related anxiety and perceptions regarding changes in risk of COVID-19 severity due to smoking. We used cross-sectional data from a population telephone survey that was conducted in Israel in the early phase of the COVID-19 pandemic (May-June 2020) and included 420 adult (age 18+) individuals who reported having ever used CCs (n = 391), nargila (n = 193), and/or electronic cigarettes (e-cigarettes)/heated tobacco products (e.g., IQOS) (n = 52). Respondents were asked about the effect that COVID-19 had on their nicotine product use (quit/reduced use, no change, increased use). We assessed changes in product use, risk perceptions, and anxiety using adjusted multinomial logistic regression analyses. Most respondents did not change their frequency of product use (CCs: 81.0%, nargila: 88.2%, e-cigarettes/IQOS: 96.8%). A small percentage either decreased use (CCs: 7.2%, nargila: 3.2%, e-cigarettes/IQOS:2.4%) or increased use (CCs:11.8%, nargila:8.6%, e-cigarettes/IQOS:+ 0.9%). 55.6% of respondents used a product in the home prior to COVID-19; but during the first lockdown COVID-19 period, a greater percentage increased (12.6%) than decreased (4.0%) their home use. Higher levels of anxiety due to COVID-19 were associated with increased home smoking (aOR = 1.59, 95% CI:1.04-2.42, p = 0.02). Many respondents believed that increased severity of COVID-19 illness was associated with CCs (62.0%) and e-cigarettes/vaping (45.3%), with uncertainty about the association being lower for CCs (20.5%) than for vaping (41.3%). While many respondents believed that nicotine product use (particularly CCs and e-cigarettes) was associated with increased risk of COVID-19 disease severity, the majority of users did not change their tobacco/nicotine use. The confusion about the relationship between tobacco use and COVID-19 calls for clear evidence-based messaging from governments. The association between home smoking and increased COVID-19-related stress suggests the need for campaigns and resources to prevent smoking in the home, particularly during times of stress.


A Quality Improvement Project to Improve Family Recognition of Medical Team Member Roles.
Previous studies have shown that inpatients and families in academic settings have a limited ability to recall either their medical team members or the roles of those members. This is an important issue for patient and family satisfaction as well as patient safety. The objective of this study was to increase families' recognition of medical team members' roles. We established a multidisciplinary quality improvement leadership team, measured family recognition of medical team members and their roles, and conducted 2 PDSA (Plan-Do-Study-Act) cycles. The first intervention was standardization of the content and delivery of our verbal team introductions to ensure inclusion of essential elements and family engagement. The second intervention was addition of an informational white board in each patient room. The prospective study included 105 families in the preintervention phase, 103 post-PDSA cycle 1, and 92 post-PDSA cycle 2. After conduction of 2 PDSA cycles, the recognition of the attending role increased from 49% to 87% (P = .000), the resident role from 39% to 73% (P = .000), and the medical student from 75% to 89% (P = .038). The multidisciplinary quality improvement model was effective in improving family recognition of the roles of attending physicians, resident physicians, and medical students. Consistent attention to engaging the families and explaining our roles as well as providing informational white boards are effective interventions to facilitate this process.


Factor XI activity and factor XI antigen in homozygous and heterozygous factor XI deficiency.
A relatively potent antiserum against highly purified, unactivated human factor XI antigen was raised in a rabbit. This antiserum, after concentration, neutralized 50% of the factor XI clotting activity of a standard normal plasma at an antiserum dilution of 1/900. The antiserum was used in a neutralization-inhibition assay to study the relation between factor XI clotting activity and factor XI antigen in plasma from ten unrelated patients with homozygous factor XI deficiency and from 12 heterozygous family members of these patients. No evidence of factor XI antigen significantly in excess of factor XI activity was found in either group. All data to date have been consistent with the hypothesis that hereditary factor XI deficiency represents a genetic disorder resulting from the absence of factor XI molecule. Severity of bleeding in factor XI deficiency could not be correlated with the level of factor XI activity or factor XI antigen.


The Recovery of Cognitive and Affective Deficiencies Linked with Chronic Osteoarthritis Pain and Implicated Pathways by Slow-Releasing Hydrogen Sulfide Treatment.
Chronic osteoarthritis pain is accompanied by several comorbidities whose treatment has not been completely resolved. The anti-inflammatory, analgesic, and antidepressant effects of slow-releasing hydrogen sulfide (H2S) donors during osteoarthritic pain have been shown, but their actions in the accompanying memory impairment and anxious-like behaviors have not yet been demonstrated. Using female mice with chronic osteoarthritic pain, the effects of natural, diallyl disulfide (DADS) or synthetic, morpholin-4-ium 4-methoxyphenyl(morpholino) phosphinodithioate dichloromethane complex (GYY4137) slow-releasing H2S donors, on associated cognitive and grip strength deficits and anxiodepressive-like behaviors, were assessed. Their effects on specific brain areas implicated in the modulation of pain and emotional responses were also determined. Results demonstrated an improvement in memory and grip strength deficits, as well as in the anxious-like behaviors associated with chronic pain in GYY4137 and/or DADS treated mice. The painkiller and antidepressant properties of both treatments were also established. Treatment with DADS and/or GYY4137 inhibited: oxidative stress in the amygdala; phosphoinositide 3-kinase overexpression in the amygdala, periaqueductal gray matter, and anterior cingulate cortex; protein kinase B activation in the amygdala and infralimbic cortex; up-regulation of inducible nitric oxide synthase in the amygdala, periaqueductal gray matter and infralimbic cortex and apoptotic responses in the amygdala. These results might explain the recovery of memory and grip strength and the inhibition of allodynia and associated anxiodepressive-like behaviors by these treatments. In conclusion, this study revealed new properties of slow-releasing H2S donors in cognitive impairment and affective disorders linked with chronic osteoarthritis pain and their effects on the central nervous system.


Urea ammoniation effects on the feeding value of guineagrass (Panicum maximum) hay.
Laboratory, digestion, and growth studies evaluated urea as a source of ammoniation for quality improvement in guineagrass (Panicum maximum) hay. In a laboratory trial, 5.0-kg portions of hay were reconstituted with water to yield final forage moisture concentrations or 25 of 40% and treated with urea at 0, 4, 6, or 8% of the forage DM, with or without urease addition. Main effects of forage moisture or urease addition did not influence (P > .10) CP or NDF concentration or in vitro OM disappearance (IVOMD) of the guineagrass hay. Hay CP concentration and IVOMD increased linearly (P < .01), whereas concentrations of hemicellulose and ADL decreased linearly (P < .05) with increasing urea level. In other experiments, round bales of hay (320 kg) were reconstituted with water to yield final forage moisture concentrations of 25 or 40% and treated with urea at 0, 4, or 6% of the forage DM. The urea solution was applied as a spray onto the cut edges of the bales, or by low pressure (10 psi) injection. Two- and three-way interactions (P < .05) existed among forage moisture concentration, urea application method, and urea level for CP and NDF concentration and IVOMD of the guineagrass hay. Greatest enhancements in these forage quality characteristics were obtained when the urea solution was sprayed onto the hay at the 25% forage moisture concentration. In two digestion and two growth trials, round bales of hay were treated with 0, 4, and 6% urea sprayed onto the hay at the 25% forage moisture level. In each growth trial, 30 St. Croix white hair castrated male sheep (Trial 1:34 +/- 5.5 kg, Trial 2: 17 +/- 3.5 kg) were allotted to six pens of five head each, resulting in two pens per treatment. In the digestion trials, six similar sheep were used in a replicated 3 x 3 Latin square design. In the digestion and growth trials, hay intake increased in a quadratic (P < .05) manner with increasing urea level. Apparent NDF and ADF digestibilities increased linearly (P < .05) with increasing urea level. Linear improvements in ADG (P < .05) and gain/feed (P < .07) were observed with increasing urea level. Urea ammoniation offers potential for improving the feeding value of tropical forages and provides an option for quality forage during the dry season.


A rapid polymerase-chain-reaction-directed sequencing strategy using a thermostable DNA polymerase from Thermus flavus.
We have developed a polymerase chain reaction (PCR)-directed sequencing strategy for rapid sequencing of DNA from crude viral or cell preparations. The basic strategy consists of two phases. In the first phase, the target DNA is amplified by symmetric PCR with low concentrations of deoxyribonucleotide triphosphate (dNTP) and oligodeoxyribonucleotide primers. This results in exponential amplification of DNA in the initial cycles, reaching a plateau by 25 cycles due to limiting concentrations of dNTP and primers. In the second phase, a small aliquot of the PCR mixture is amplified without any purification, by asymmetric PCR in the presence of a 5'-labeled primer and one of the four dideoxyribonucleotide triphosphates. This results in the accumulation of single-stranded DNA products that are terminated at specific points by incorporation of the appropriate dideoxyribonucleotide monophosphate. The products are then analyzed by electrophoresis on a sequencing gel followed by autoradiography. The PCR conditions are optimized to generate sequence ladders of several hundred nucleotides starting from as low as 100 copies of bacteriophage or bacterial genome in one to two days.


Histological evaluation of poly(L-lactide/epsilon-caprolactone) membrane implanted subcutaneously in rats.
Poly(L-lactide/epsilon-caprolactone) [P(L-LA/epsilon-CL)] 50/50 membranes is an absorbable membrane which has been developed recently for possible use in tissue engineering. To look at histological tissue reactions to the membrane and its behaviour upon its in vivo implantation in rats. P(L-LA/e-CL) 50/50 membranes, 0.4 mm thick, were implanted subcutaneously in the dorsal neck of 20 Wistar rats. The rats were followed-up for 1, 3, 6, 12 and 28 months. After sacrifice, subcutaneous tissues with implants were taken as specimens, inspected for any gross abnormality and histological examination was carried out. No significant macroscopic changes were noticed except for two implants that were grossly rounded. Histologically, the membranes had undergone cracking, fragmentation and progressive degradation. Cracks appeared initially at the periphery of the implant. Large cracks progressed mainly in longitudinal planes. Cracks occurred in the middle of implants and led to membrane bending in some cases. "Neomembrane" formation, comprised of fibrous tissue and the implant, was seen. The foreign-body reaction to the membrane involved macrophages and foreign-body giant cells. In one case an acute type of polymorphonuclear cell reaction was observed and in a second case a predominantly lymphocytic reaction was seen at three months. They were thought to be the results of infection. No other adverse effects were seen. P(L-LA/e-CL) 50/50 membrane was found to be biocompatible when implanted subcutaneously in rats. It was degraded to a great extent but not completely in 28 months.


Body composition of female wheelchair athletes.
Wheelchair users undergo changes in body composition as a result of disability. In this study the distribution of bone mineral, lean and fat mass was assessed in highly-trained female wheelchair athletes and a reference group by dual-energy X-ray absorptiometry (DXA). The transferability of anthropometric equations commonly used in female groups was examined in order to establish a suitable field method of body composition assessment. The DXA total-body results indicated no difference between groups, but segmental analyses uncovered regional differences. The wheelchair athletes had greater BMD (p=0.088), more lean mass (p<0.001) and a lower percent fat (p=0.050) in their arms. The reverse was true of the legs (p< or =0.001). The trunk as a whole did not differ between groups. In general, the anthropometric equations showed a lack of transferability to the wheelchair group and tended to underestimate total percent body fat. Anthropometric measures such as body mass index (BMI) and waist girth showed strong correlations with body fat in the wheelchair group (BMI: r=0.90, p=0.001; waist: r=0.83, p=0.001), but weaker results in the reference group. It is recommended that specific anthropometric equations be developed for use in the absence of a 'gold standard' measure of body composition such as DXA.


Improving brain injury cognitive rehabilitation by personalized telerehabilitation services: Guttmann neuropersonal trainer.
Cognitive rehabilitation aims to remediate or alleviate the cognitive deficits appearing after an episode of acquired brain injury (ABI). The purpose of this work is to describe the telerehabilitation platform called Guttmann Neuropersonal Trainer (GNPT) which provides new strategies for cognitive rehabilitation, improving efficiency and access to treatments, and to increase knowledge generation from the process. A cognitive rehabilitation process has been modeled to design and develop the system, which allows neuropsychologists to configure and schedule rehabilitation sessions, consisting of set of personalized computerized cognitive exercises grounded on neuroscience and plasticity principles. It provides remote continuous monitoring of patient's performance, by an asynchronous communication strategy. An automatic knowledge extraction method has been used to implement a decision support system, improving treatment customization. GNPT has been implemented in 27 rehabilitation centers and in 83 patients' homes, facilitating the access to the treatment. In total, 1660 patients have been treated. Usability and cost analysis methodologies have been applied to measure the efficiency in real clinical environments. The usability evaluation reveals a system usability score higher than 70 for all target users. The cost efficiency study results show a relation of 1-20 compared to face-to-face rehabilitation. GNPT enables brain-damaged patients to continue and further extend rehabilitation beyond the hospital, improving the efficiency of the rehabilitation process. It allows customized therapeutic plans, providing information to further development of clinical practice guidelines.


Effect of rapid influenza diagnostic tests on patient management in an emergency department.
We evaluated the effect of rapid influenza diagnostic tests (RIDTs) on patient management in an emergency department for 3 years after 2009, and also identified factors associated with the choice of treatment for patients with influenza-like illnesses. The study period consisted of three influenza epidemic seasons. Patients older than 15 years who underwent RIDTs in the emergency department and were then discharged without admission were included. A total of 453 patients were enrolled, 114 of whom had positive RIDT results. Antiviral medication was prescribed to 103 patients (90.4%) who had positive RIDT results, while 1 patient (0.3%) who tested negative was treated with antivirals (P<0.001). Conservative care was administered to 11 RIDT-positive patients (9.6%) and 244 RIDT-negative patients (72.0%) (P<0.001). Symptom onset in less than 48 hours, being older than 65 years, and the presence of comorbidities were not associated with the administration of antiviral therapy. RIDT results had a critical effect on physician decision-making regarding antiviral treatment for patients with influenza-like illnesses in the emergency department. However, symptom onset in less than 48 hours, old age, and comorbidities, which are all indications for antiviral therapy, were not found to influence the administration of antiviral treatment.


[Peculiarities of theophylline and prolactin interaction and calcium fluctuation from intracellular stores of porcine oocytes in the presence of estradiol].
Using a fluorescent dye chlortetracycline, a study was made of the effect of estradiol on the interaction of theophylline and prolactin in the course of Ca2+ exit from intracellular stores of pig oocytes, isolated from ovaries at the stage of follicle growth. It is shown that in the presence of estradiol, prolactin does not stimulate Ca2+ exit from intracellular stores of pig oocytes. The action of theophylline similarly does not stimulate Ca2+ exit. Unlike, a joint effect of theophylline and prolactin on pig oocytes in the presence estradiol stimulated Ca2+ exit from intracellular stores of pig oocytes. These data demonstrated the influence of estradiol on theophylline and prolactin stimulated Ca2+ exit from intracellular stores of pig oocytes.


Interactions of polybrominated diphenyl ethers with the aryl hydrocarbon receptor pathway.
Polybrominated diphenyl ethers (PBDEs) are brominated flame retardants that have been in use as additives in various consumer products. Structural similarities of PBDEs with other polyhalogenated aromatic hydrocarbons that show affinity for the aryl hydrocarbon receptor (AhR), such as some polychlorinated biphenyls, raised concerns about their possible dioxin-like properties. We studied the ability of environmentally relevant PBDEs (BDE-47, -99, -100, -153, -154, and -183) and the "planar" congener BDE-77 to bind and/or activate the AhR in stably transfected rodent hepatoma cell lines with an AhR-responsive enhanced green fluorescent protein (AhR-EGFP) reporter gene (H1G1.1c3 mouse and H4G1.1c2 rat hepatoma). 7-Ethoxyresorufin-O-deethylation (EROD) was used as a marker for CYP1A1 activity. Dose- and bromination-specific inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced responses was measured by their ability to inhibit the induction of AhR-EGFP expression and EROD activity. Individual exposure to these PBDEs did not result in any increase in induction of AhR-EGFP or CYP1A1 activity. The lower brominated PBDEs showed the strongest inhibitory effect on TCDD-induced activities in both cell lines. While the highest brominated PBDE tested, BDE-183, inhibited EROD activity, it did not affect the induction of AhR-EGFP expression. Similar findings were observed after exposing stably transfected human hepatoma (xenobiotic response element [XRE]-HepG2) cells to these PBDEs, resulting in a small but statically significant agonistic effect on XRE-driven luciferase activity. Co-exposure with TCDD resulted again in antagonistic effects, confirming that the inhibitory effect of these PBDEs on TCDD-induced responses was not only due to direct interaction at receptor level but also at DNA-binding level. This antagonism was confirmed for BDE-99 in HepG2 cells transiently transfected with a Gal4-AhR construct and the corresponding Gal4-Luc reporter gene. In addition, a chromatin immunoprecipitation assay further confirmed that BDE-99 could bind to the AhR and activate the AhR nuclear translocation and dioxin responsive element (DRE) binding in the context of the CYP1A1 promoter. However, the transactivation function of the BDE-99-activated AhR seems to be very weak. These combined results suggest that PBDEs do bind but not activate the AhR-AhR nuclear translocator protein-XRE complex.


Two splice variants of Golgi-microtubule-associated protein of 210 kDa (GMAP-210) differ in their binding to the cis-Golgi network.
GMAP-210 (Golgi-microtubule-associated protein of 210 kDa) is a peripheral Golgi protein that interacts with the minus end of microtubules through its C-terminus and with cis-Golgi network membranes through its N-terminus; it participates in the maintenance of the structural integrity of the Golgi apparatus [Infante, Ramos-Morales, Fedriani, Bornens and Rios (1999) J. Cell Biol. 145, 83--98]. We report here the cloning of a new isoform of GMAP-210 that lacks amino acid residues 105--196. On the basis of the analysis of the gmap-210 genomic sequence, we propose that the small isoform, GMAP-200, arises from alternative splicing of exon 4 of the primary transcript. Overexpression of GMAP-200 induces perturbations in both the Golgi apparatus and the microtubule network that are similar to those previously reported for GMAP-210 overexpression. We show that both isoforms are able to oligomerize under overexpression conditions. Analysis in vitro and in vivo, with the green fluorescent protein as a marker, reveals that the binding of the N-terminal domain of GMAP-200 to the cis-Golgi network membranes is lower than that of the N-terminal domain of GMAP-210. Implications for the regulation of interaction between the cis-Golgi network and microtubules are discussed.


Evolution of Genetic Variance during Adaptive Radiation.
Genetic correlations between traits can concentrate genetic variance into fewer phenotypic dimensions that can bias evolutionary trajectories along the axis of greatest genetic variance and away from optimal phenotypes, constraining the rate of evolution. If genetic correlations limit adaptation, rapid adaptive divergence between multiple contrasting environments may be difficult. However, if natural selection increases the frequency of rare alleles after colonization of new environments, an increase in genetic variance in the direction of selection can accelerate adaptive divergence. Here, we explored adaptive divergence of an Australian native wildflower by examining the alignment between divergence in phenotype mean and divergence in genetic variance among four contrasting ecotypes. We found divergence in mean multivariate phenotype along two major axes represented by different combinations of plant architecture and leaf traits. Ecotypes also showed divergence in the level of genetic variance in individual traits and the multivariate distribution of genetic variance among traits. Divergence in multivariate phenotypic mean aligned with divergence in genetic variance, with much of the divergence in phenotype among ecotypes associated with changes in trait combinations containing substantial levels of genetic variance. Overall, our results suggest that natural selection can alter the distribution of genetic variance underlying phenotypic traits, increasing the amount of genetic variance in the direction of natural selection and potentially facilitating rapid adaptive divergence during an adaptive radiation.


Pharmacologic therapeutics for cardiac reperfusion injury.
Cardiovascular disease is the leading cause of morbidity and mortality in industrial societies, with myocardial infarction as the primary assassin. Pharmacologic agents, including the myocardial cell membrane receptor agonists adenosine, bradykinin/angiotensin-converting enzyme inhibitors, opioids and erythropoietin or the mixed cell membrane and intracellular agonists, glucose insulin potassium, and volatile anesthetics, either clinically or experimentally reduce the extent of myocardial injury when administered just prior to reperfusion. Agents that specifically target proteins, transcription factors or ion channels, including PKC agonists/antagonists, PPAR, Phosphodiesterase-5 inhibitors, 3-Hydroxy-3-methyl glutaryl coenzyme A reductase and the ATP-dependent potassium channel are also promising. However, no agent has been specifically approved to reduce reperfusion injury clinically. In this review, we will discuss the advantages and limitations of agents to combat reperfusion injury, their market development status and findings reported in both clinical and preclinical studies. The molecular pathways activated by these agents that preserve myocardium from reperfusion injury, which appear to commonly involve glycogen synthase kinase 3beta and mitochondrial permeability transition pore inhibition, are also described.


Bone Morphogenetic Protein Antagonist Gremlin-1 Increases Myofibroblast Transition in Dermal Fibroblasts: Implications for Systemic Sclerosis.
Systemic Sclerosis is an autoimmune connective tissue disease which results in fibrosis of the skin and lungs. The disease is characterized by activation of myofibroblasts but what governs this is unknown. Gremlin-1 is a BMP antagonist that is developmentally regulated and we sought to investigate its role in Systemic Sclerosis. Dermal fibroblasts were transfected with Grem1pcDNA3.1 expression vectors or empty vectors. Various markers of myofibroblasts were measured at the mRNA and protein levels. Scratch wound assays were also performed. Media Transfer experiments were performed to evaluate cytokine like effects. Various inhibitors of TGF-β signaling and MAPK signaling were used post-transfection. siRNA to Gremlin-1 in SSc dermal fibroblasts were performed to evaluate the role of Gremlin-1. Different cytokines were incubated with fibroblasts and Gremlin-1 measured. Bleomycin was used as model of fibrosis and immunohistochemistry performed. Overexpression of Gremlin-1 was achieved in primary dermal fibroblasts and lead to activation of quiescent cells to myofibroblasts indicated by collagen and α-Smooth muscle actin. Overexpression also led to functional effects. This was associated with increased TGF-β1 levels and SBE luciferase activity but not increased Thrombospondin-1 expression. Inhibition of Gremlin-1 overexpression cells with antibodies to TGF-β1 but not isotype controls led to reduced collagen and various TGF-β pathway chemical inhibitors also led to reduced collagen levels. In SSc cells siRNA mediated reduction of Gremlin-1 reduced collagen expression and CTGF gene and protein levels in these cells. IL-13 did not lead to elevated Gremlin-1 expression nor did IL-11. Gremlin-1 was elevated in an animal model of fibrosis compared to NaCl-treated mice. Gremlin-1 is a key regulator of myofibroblast transition leading to enhanced ECM deposition. Strategies that block Gremlin-1 maybe a possible therapeutic target in fibrotic diseases such as SSc.


Calibration transfer from dispersive instruments to handheld spectrometers.
The Foss NIRSystem 6500 is one of the most commonly used laboratory instruments in agriculture and in particular in feed. New technological developments include micro-electro mechanical system (MEMS) technology, used in miniature handheld instruments such as the Polychromix Phazir spectrometer that are increasingly required for on-site analysis. The objective of this study was to assess the potential of a calibration transfer from the Foss NIRSystem 6500 to the Polychromix Phazir. The results show that good calibration models were obtained for various feed properties (fat, fiber, protein, and starch) developed on a Foss NIRSystem 6500, based on a spectral database of 9164 samples transferred to a Polychromix Phazir handheld spectrometer.


Determinants of adherence to wrap-around care in child and family services.
The aim of this study is to understand the determinants of adherence to wrap-around care (WAC) by professional care providers working in child and family services. WAC is a care coordination method targeting families with complex needs. The core components of WAC involve activating family members and the social network, integrating the care provider network, and assessing, planning and evaluating the care process. WAC was introduced in the Netherlands using two approaches: the network approach (NA) and the team approach (TA). A cross-sectional study was conducted using a digital questionnaire targeted at care providers. After imputation of missing data, univariate and multilevel regression analyses were conducted to study the associations between adherence to the core components of WAC, the determinants of adherence and background characteristics. In total 145 out of 275 care providers (52.7%) responded to the questionnaire. Multilevel regression analysis showed that self-efficacy of the care providers and the way WAC is organised (NA versus TA region) were significantly associated with adherence to core components of WAC. Self-efficacy was significantly associated with all WAC core components (activating family members and the social network: β (95% confidence interval, CI) = .27(.04-.50), integrating the network of care providers: β (95% CI) = .27(.05-.50) and assessing, planning and evaluating the care process: β (95% CI) = .30(.08-.52)). The way WAC is organised was significantly associated to two core components (activating family members and the social network: β (95% CI) = .18(0.1-.37) and integrating the network of care providers: β (95% CI) = .25(.09-.42)). The way WAC is organised and the self-efficacy of care providers who use WAC are factors that are relevant for the redesign of the strategy for introducing WAC. Longitudinal research into the predictive value of determinants of adherence to WAC is advised.


Family Quality of Life During the Transition to Adulthood for Individuals With Intellectual Disability and/or Autism Spectrum Disorders.
Recognizing the prominent role of parents in supporting their children with intellectual and developmental disabilities (IDD), improving quality of life for these families is an essential endeavor. While much attention has focused on the experiences of families with younger children with IDD, little is known about the family quality of life (FQOL) among families with transition-age youth and young adults. We examined the FQOL ratings of 425 parents with a child between 13-21 years of age with intellectual disability or autism to understand FQOL and the factors that may shape it. Overall satisfaction with FQOL was somewhat high for this sample, with some variability across domains. Higher FQOL ratings were predicted by lower frequency of challenging behaviors, lower support needs, and higher strength of parental religious faith. We present recommendations for research and practice focused on promoting quality of life during the transition period.


Immunotherapy of allergic bronchopulmonary aspergillosis: a clinical and experimental approach.
Allergic bronchopulmonary aspergillosis (ABPA) is a severe allergic pulmonary complication caused by the saprophytic fungus Aspergillus fumigatus. The present review examines the pathogenesis of this disease describing in detail the role of innate and acquired immunity in the induction of sensitivity to A.fumigatus. Different approaches in developing specific immunotherapeutic treatments such as induction of anergy, regulatory cells, a switch from Th2 to Th1 type of immune response, CpG and genetic immunization and the usage of altered peptides or modified allergens are critically examined.


Evaluation of brief screening tools for neurocognitive impairment in HIV/AIDS: a systematic review of the literature.
To systematically review literature on brief screening tools used to detect and differentiate between normal cognition and neurocognitive impairment and HIV-associated neurocognitive disorders (HANDs) in adult populations of persons with HIV. A formal systematic review. We searched six electronic databases in 2011 and contacted experts to identify relevant studies published through May 2012. We selected empirical studies that focused on evaluating brief screening tools (<20 min) for neurocognitive impairment in persons with HIV. Two reviewers independently reviewed retrieved literature for potential relevance and methodological quality. Meta-analyses were completed on screening tools that had sufficient data. Fifty-one studies met inclusion criteria; we focused on 31 studies that compared brief screening tools with reference tests. Within these 31 studies, 39 tools were evaluated and 67% used a comprehensive neuropsychological battery as a reference. The majority of these studies evaluated HIV-associated dementia (HAD). Meta-analyses demonstrated that the HIV Dementia Scale (HDS) has poor pooled sensitivity (0.48) and the International HIV Dementia Scale (IHDS) has moderate pooled sensitivity (0.62) in detecting a range of cognitive impairment. Five newer screening tools had relatively good sensitivities (>0.70); however, none of the tools differentiated HAND conditions well enough to suggest broader use. There were significant methodological shortcomings noted in most studies. HDS and IHDS perform well to screen for HAD but poorly for milder HAND conditions. Further investigation, with improved methodology, is required to understand the utility of newer screening tools for HAND; further tools may need to be developed for milder HAND conditions.


Microbial community composition of a multi-stage moving bed biofilm reactor and its interaction with kinetic model parameters estimation.
Microbial community diversity determines the function of each chamber of multi-stage moving bed biofilm reactor (MBBR) systems. How the microbial community data can be further used to serve wastewater treatment process modelling and optimization has been rarely studied. In this study, a MBBR system was set up to investigate the microbial community diversity of biofilm in each functional chamber. The compositions of microbial community of biofilm from different chambers of MBBR were quantified by high-throughput sequencing. Significantly higher proportion of autotrophs were found in the second aerobic chamber (15.4%), while 4.3% autotrophs were found in the first aerobic chamber. Autotrophs in anoxic chamber were negligible. Moreover, ratios of active heterotrophic biomass and autotrophic biomass (XH/XA) were obtained by performing respiration tests. By setting heterotroph/autotroph ratios obtained from sequencing analysis equal to XH/XA, a novel approach for kinetic model parameters estimation was developed. This work not only investigated microbial community of MBBR system, but also it provided an approach to make further use of molecular microbiology analysis results.


A comparison of the genomes of bovine herpesvirus type 1 and pseudorabies virus.
The DNA sequence homology between bovine herpesvirus type 1 (BHV-1) and pseudorabies virus (PRV) was examined. Reciprocal cross-hybridization of viral DNA labelled by nick translation to Southern blots of restriction endonuclease-digested DNA detected homologous sequences dispersed throughout the genomes of the two viruses. The DNA-DNA hybrids formed were stable under high-stringency wash conditions. Sequences of a 32P-labelled PRV DNA fragment probe were found to hybridize only to a specific region of the BHV-1 genome, suggesting that the detected sequence homology was not due to fortuitous hybridization of G + C-rich sequences. As measured by liquid reassociation kinetics the homology between these two viruses was approximately 8%.


Venture from the Interior-Herpesvirus pUL31 Escorts Capsids from Nucleoplasmic Replication Compartments to Sites of Primary Envelopment at the Inner Nuclear Membrane.
Herpesviral capsid assembly is initiated in the nucleoplasm of the infected cell. Size constraints require that newly formed viral nucleocapsids leave the nucleus by an evolutionarily conserved vescular transport mechanism called nuclear egress. Mature capsids released from the nucleoplasm are engaged in a membrane-mediated budding process, composed of primary envelopment at the inner nuclear membrane and de-envelopment at the outer nuclear membrane. Once in the cytoplasm, the capsids receive their secondary envelope for maturation into infectious virions. Two viral proteins conserved throughout the herpesvirus family, the integral membrane protein pUL34 and the phosphoprotein pUL31, form the nuclear egress complex required for capsid transport from the infected nucleus to the cytoplasm. Formation of the nuclear egress complex results in budding of membrane vesicles revealing its function as minimal virus-encoded membrane budding and scission machinery. The recent structural analysis unraveled details of the heterodimeric nuclear egress complex and the hexagonal coat it forms at the inside of budding vesicles to drive primary envelopment. With this review, I would like to present the capsid-escort-model where pUL31 associates with capsids in nucleoplasmic replication compartments for escort to sites of primary envelopment thereby coupling capsid maturation and nuclear egress.


Risky Business? A Year-Long Study of Adult Voluntary Admissions Who Leave Psychiatric in-Patient Care without Informing Staff.
Patients leaving inpatient psychiatric care without informing staff can have serious consequences for patient welfare, their families and staff. Even in cases where there are no serious outcomes and patients return, treatment can be interrupted and staff morale compromised. Given these potential deleterious effects, it is important to have an in-depth understanding of why people leave when they do. The aim of this study is to examine the profile of service-users who leave in-patient psychiatric care without notifying staff and to determine their reasons for doing so, what they did when they left the hospital setting and reasons for returning. This is a prospective descriptive study of service users leaving without notifying staff from a national mental health service in Ireland. It investigated the frequency, timing, and motivations of these absconding events among a sample of in-patients over a 12-month period. This year-long review identified 75 episodes of leaving without notifying staff, 55 of which met this studies criteria. 22% of episodes were from a locked unit, 58.2% were repeated episodes. 89% of service users returned to hospital following the episode and 11% were discharged against medical advice. Although no fatalities, three service users were seen in an emergency department as a result of activity during leave. Upon return from leave, all service users were reviewed and their care plans were updated where necessary.


Assessment of hip abductor muscle strength. A validity and reliability study.
Hip abductors are the most important muscles around the hip joint. It is therefore essential to assess their function in a valid and reliable way. Since the optimal body posture for the assessment of hip abductor strength is unknown, we tested the validity and reliability of unilateral hip abductor strength assessment in three different body positions. We hypothesized that the validity would be better in the side-lying position because of the consistent stabilization of the contralateral (untested) hip. Sixteen healthy subjects participated in two identical testing sessions. Unilateral isometric hip abductor muscle strength was measured, with use of a stabilized commercial dynamometer, with the subject in the side-lying, supine, and standing positions. Construct validity was based on concomitant recordings of gluteus medius electromyographic activity from the tested and contralateral hips. The body position permitting greater muscle activation and abductor strength on the tested hip, while minimizing muscle activation in the contralateral hip (that is, lower contralateral-to-tested electromyographic ratio), was considered the most valid. Coefficients of variation, the Bland and Altman limits of agreement, and intraclass correlation coefficients were calculated to determine test-retest reliability of hip abductor strength. Maximal hip abductor strength was significantly higher in the side-lying position compared with the standing and supine positions (p < 0.05). The contralateral-to-tested electromyographic ratio for the side-lying position was significantly lower than that for the supine and the standing position (p < 0.01). Test-retest reliability of strength measurements in terms of coefficients of variation (3.7% for side-lying, 6.1% for supine, and 4.2% for standing) and limits of agreement (+/-6.9% for side-lying, +/-8.4% for supine, and +/-7.5% for standing) was better in the side-lying position. All intraclass correlation coefficients were high to moderate (0.90 for side-lying, 0.83 for supine, and 0.88 for standing). The side-lying body position offers the most valid and reliable assessment of unilateral hip abductor strength.


Fluorescence recovery assay for the detection of protein-DNA binding.
We report a novel and straightforward fluorescence recovery assay which enables the detection of protein-DNA interactions and simultaneously determines relative binding affinities of sequence-specific DNA-binding proteins for a variety of DNA sequences in a multiplexed format. The detection of protein-DNA binding is accomplished by monitoring fluorescence recovery during exonuclease digestion of DNA sequences that are modified with fluorophore-quencher pairs. Retardation of fluorescence recovery occurs with binding of the protein to the putative DNA binding element, which arrests exonuclease digestion. The assay detects protein-DNA binding in a homogeneous solution simply, quickly, and reliably without using radioisotopes. Multiplexing is possible by labeling different DNA sequences with spectrally distinct dyes, allowing simultaneous analysis of experimental and control binding reactions in the same mixture.


A Method to Experimentally Estimate the Conductivity of Chronic Stroke Lesions: A Tool to Individualize Transcranial Electric Stimulation.
The inconsistent response to transcranial electric stimulation in the stroke population is attributed to, among other factors, unknown effects of stroke lesion conductivity on stimulation strength at the targeted brain areas. Volume conduction models are promising tools to determine optimal stimulation settings. However, stroke lesion conductivity is often not considered in these models as a source of inter-subject variability. The goal of this study is to propose a method that combines MRI, EEG, and transcranial stimulation to estimate the conductivity of cortical stroke lesions experimentally. In this simulation study, lesion conductivity was estimated from scalp potentials during transcranial electric stimulation in 12 chronic stroke patients. To do so, first, we determined the stimulation configuration where scalp potentials are maximally affected by the lesion. Then, we calculated scalp potentials in a model with a fixed lesion conductivity and a model with a randomly assigned conductivity. To estimate the lesion conductivity, we minimized the error between the two models by varying the conductivity in the second model. Finally, to reflect realistic experimental conditions, we test the effect rotation of measurement electrode orientation and the effect of the number of electrodes used. We found that the algorithm converged to the correct lesion conductivity value when noise on the electrode positions was absent for all lesions. Conductivity estimation error was below 5% with realistic electrode coregistration errors of 0.1° for lesions larger than 50 ml. Higher lesion conductivities and lesion volumes were associated with smaller estimation errors. In conclusion, this method can experimentally estimate stroke lesion conductivity, improving the accuracy of volume conductor models of stroke patients and potentially leading to more effective transcranial electric stimulation configurations for this population.


Reliability and validity of the Acute Care Index of Function for patients with neurologic impairment.
The Acute Care Index of Function (ACIF), developed to standardize the assessment of functional status in patients with acute neurologic deficits, was tested for interrater reliability and concurrent validity. Interrater reliability was tested on 91 patients in a five-week study. After a two-week training period, six physical therapists were paired so that each therapist was paired with each other therapist for one week. All patients were rated weekly with the ACIF by a pair of physical therapists. A total of 166 assessments were conducted. Concurrent validity was tested using descriptions of function of 10 hypothetical patients. Each patient description was assigned a score based on the ACIF. Ten experienced therapists were then asked to rank order the functional status of each patient based on the patient's description. The average of the physical therapists' rankings was correlated with the ranking determined with the ACIF scores. Reliability data were analyzed using Cohen's weighted Kappa. The weighted Kappa values ranged from .60 to .98. Validity data were analyzed with the Spearman rank-order correlation coefficient (rs = .81, p less than .01). The results of this study indicate that the ACIF is reliable between raters and valid in its ability to rank order functional status for patients with acute neurologic deficits.


POEMS Syndrome: Real World Experience in Diagnosis and Systemic Therapy - 108 Patients Multicenter Analysis.
POEMS syndrome, a rare plasma cell disorder, is challenging both in the diagnostic and therapeutic management. We present real word retrospective analysis of 108 cases analyzing clinical features and therapeutic modes. We compare our results with the available literature. This is the first description with such wide use of proteasome inhibitors in first line treatment. POEMS (Polyneuropathy, organomegaly, endocrinopathy, M-protein, skin changes) syndrome is a rare and challenging plasma cell disorder, both in the diagnostic and therapeutic management of the disease. Currently, the literature on POEMS is sparse with most evidence being case reports and small case studies. We present a retrospective real world experience of 108 patients with POEMS. We analyzed the clinical features and therapeutic interventions. Regarding clinical features, our findings demonstrated that skin lesions, thrombocythemia and polycythemia were present less frequently than reported previously. Regarding clinical interventions, this is one of the largest analyses of front line treatment in POEMS and the first one to include frequent utilization of proteasome inhibitors (37%). Bortezomib monotherapy was the most effective therapy achieving complete remission/very good partial remissions (CR/VGPR) in 69% of patients. Thirty percent of patients proceeded to planned autologous stem cell transplant (ASCT) as part of the front-line treatment resulting in statistically superior progression-free (PFS) and overall survival (OS) compared to non-ASCT treated patients (P= .003). In multivariate analysis, anemia, thrombocytopenia, and as age over 60 were associated with a negative impact on patient outcomes.


A conservative alternative to lower posterior bridges replacing mandibular first molars.
The permanent first molars erupt at the age of six years old. These teeth are most prone to dental caries and are often extracted at an early age because of dental decay. The conventional approach to replace these extracted teeth include fixed bridge, removable side plate and removable partial denture. In suitable cases, spaces created by the missing permanent first molars can be closed by the use of orthodontic appliances thereby avoiding the unnecessary sacrifice of sound tooth structure or the discomfort of wearing removable partial prosthesis.


Is adipose tissue a place for Mycobacterium tuberculosis persistence?
Mycobacterium tuberculosis, the etiological agent of tuberculosis (TB), has the ability to persist in its human host for exceptionally long periods of time. However, little is known about the location of the bacilli in latently infected individuals. Long-term mycobacterial persistence in the lungs has been reported, but this may not sufficiently account for strictly extra-pulmonary TB, which represents 10-15% of the reactivation cases. We applied in situ and conventional PCR to sections of adipose tissue samples of various anatomical origins from 19 individuals from Mexico and 20 from France who had died from causes other than TB. M. tuberculosis DNA could be detected by either or both techniques in fat tissue surrounding the kidneys, the stomach, the lymph nodes, the heart and the skin in 9/57 Mexican samples (6/19 individuals), and in 8/26 French samples (6/20 individuals). In addition, mycobacteria could be immuno-detected in perinodal adipose tissue of 1 out of 3 biopsy samples from individuals with active TB. In vitro, using a combination of adipose cell models, including the widely used murine adipose cell line 3T3-L1, as well as primary human adipocytes, we show that after binding to scavenger receptors, M. tuberculosis can enter within adipocytes, where it accumulates intracytoplasmic lipid inclusions and survives in a non-replicating state that is insensitive to the major anti-mycobacterial drug isoniazid. Given the abundance and the wide distribution of the adipose tissue throughout the body, our results suggest that this tissue, among others, might constitute a vast reservoir where the tubercle bacillus could persist for long periods of time, and avoid both killing by antimicrobials and recognition by the host immune system. In addition, M. tuberculosis-infected adipocytes might provide a new model to investigate dormancy and to evaluate new drugs for the treatment of persistent infection.


Impact of Ichthyophonus infection on spawning success of Yukon River Chinook salmon Oncorhynchus tshawytscha.
We examined the impacts of Ichthyophonus infection on spawning success of Yukon River Chinook salmon Oncorhynchus tshawytscha at spawning grounds of the Chena and Salcha Rivers, Alaska, USA. During the period 2005 to 2006, 1281 salmon carcasses (628 male, 652 female) were collected throughout the spawning season and from the entire spawning reaches of the Chena and Salcha Rivers. For each fish, infection status was determined by culture method and visual inspection of lesions of heart tissue as uninfected (culture negative), infected without lesions (culture positive with no visible lesions), and infected with lesions (culture positive with visible lesions), and spawning status was determined by visually inspecting the percentage of gametes remaining as full-spawned (<10%), partial-spawned (10-50%), and unspawned (>50%). Among the 3 groups, the proportion of full-spawned (i.e. spawning success) females was lower for those infected without lesions (69%) than those uninfected (87%) and infected with lesions (86%), but this did not apply to males (uninfected 42%, infected without lesions 38%, infected with lesions 41%). At the population level, the combined (infected and uninfected) proportion of female spawning success was 86%, compared to 87% when all females were assumed uninfected. These data suggest that while Ichthyophonus infection slightly reduces spawning success of infected females, its impact on the spawning population as a whole appears minimal.


Impact of IQ on the diagnostic yield of chromosomal microarray in a community sample of adults with schizophrenia.
Schizophrenia is a severe psychiatric disorder associated with IQ deficits. Rare copy number variations (CNVs) have been established to play an important role in the etiology of schizophrenia. Several of the large rare CNVs associated with schizophrenia have been shown to negatively affect IQ in population-based controls where no major neuropsychiatric disorder is reported. The aim of this study was to examine the diagnostic yield of microarray testing and the functional impact of genome-wide rare CNVs in a community ascertained cohort of adults with schizophrenia and low (< 85) or average (≥ 85) IQ. We recruited 546 adults of European ancestry with schizophrenia from six community psychiatric clinics in Canada. Each individual was assigned to the low or average IQ group based on standardized tests and/or educational attainment. We used rigorous methods to detect genome-wide rare CNVs from high-resolution microarray data. We compared the burden of rare CNVs classified as pathogenic or as a variant of unknown significance (VUS) between each of the IQ groups and the genome-wide burden and functional impact of rare CNVs after excluding individuals with a pathogenic CNV. There were 39/546 (7.1%; 95% confidence interval [CI] = 5.2-9.7%) schizophrenia participants with at least one pathogenic CNV detected, significantly more of whom were from the low IQ group (odds ratio [OR] = 5.01 [2.28-11.03], p = 0.0001). Secondary analyses revealed that individuals with schizophrenia and average IQ had the lowest yield of pathogenic CNVs (n = 9/325; 2.8%), followed by those with borderline intellectual functioning (n = 9/130; 6.9%), non-verbal learning disability (n = 6/29; 20.7%), and co-morbid intellectual disability (n = 15/62; 24.2%). There was no significant difference in the burden of rare CNVs classified as a VUS between any of the IQ subgroups. There was a significantly (p=0.002) increased burden of rare genic duplications in individuals with schizophrenia and low IQ that persisted after excluding individuals with a pathogenic CNV. Using high-resolution microarrays we were able to demonstrate for the first time that the burden of pathogenic CNVs in schizophrenia differs significantly between IQ subgroups. The results of this study have implications for clinical practice and may help inform future rare variant studies of schizophrenia using next-generation sequencing technologies.


Psychophysiological differences in identical twins discordant for schizophrenia: a critical index for the onset of schizophrenia.
It has been hypothesized that not only genetic but also environmental factors contribute to the onset of schizophrenia. We therefore conducted psychophysiological studies on a pair of identical twins discordant for schizophrenia, to differentiate non-genetic from genetic indexes possibly associated with this disease. The affected and unaffected twins were 28 year-old females. The affected twin was diagnosed as having schizophrenia based on the Diagnosis and Statistical Manual of Mental Disorders, third edition revised (DMS-III-R), whereas the unaffected twin had no psychiatric disorders. The brain potentials evoked by visual stimulation (visual event-related potential [visual ERP]) were recorded. The components of the visual ERP, which are believed to reflect pattern cognition, semantic processing and the failure to use preceding word information, were compared with normal subjects. Both twins showed abnormal semantic processing and similar failure to use preceding word information. Abnormality of semantic processing was marked in the affected twin. These results indicate that failure to use preceding word information might reflect only genetic factors, whereas abnormal semantic processing might reflect both genetic and non-genetic factors because the affected twin was considered to show accelerated deterioration after the disease onset. However, only the affected twin showed abnormal pattern cognition, which might be attributable to non-genetic factors such as an acquired vulnerability to schizophrenia. We suggest that the impairment of pattern cognition evaluated by visual ERP may be a critical index for the onset of schizophrenia.


Generation of tissue constructs for cardiovascular regenerative medicine: from cell procurement to scaffold design.
The ability of the human body to naturally recover from coronary heart disease is limited because cardiac cells are terminally differentiated, have low proliferation rates, and low turn-over rates. Cardiovascular tissue engineering offers the potential for production of cardiac tissue ex vivo, but is currently limited by several challenges: (i) Tissue engineering constructs require pure populations of seed cells, (ii) Fabrication of 3-D geometrical structures with features of the same length scales that exist in native tissue is non-trivial, and (iii) Cells require stimulation from the appropriate biological, electrical and mechanical factors. In this review, we summarize the current state of microfluidic techniques for enrichment of subpopulations of cells required for cardiovascular tissue engineering, which offer unique advantages over traditional plating and FACS/MACS-based enrichment. We then summarize modern techniques for producing tissue engineering scaffolds that mimic native cardiac tissue.


Flavonoids and Auxin Transport Inhibitors Rescue Symbiotic Nodulation in the Medicago truncatula Cytokinin Perception Mutant cre1.
Initiation of symbiotic nodules in legumes requires cytokinin signaling, but its mechanism of action is largely unknown. Here, we tested whether the failure to initiate nodules in the Medicago truncatula cytokinin perception mutant cre1 (cytokinin response1) is due to its altered ability to regulate auxin transport, auxin accumulation, and induction of flavonoids. We found that in the cre1 mutant, symbiotic rhizobia cannot locally alter acro- and basipetal auxin transport during nodule initiation and that these mutants show reduced auxin (indole-3-acetic acid) accumulation and auxin responses compared with the wild type. Quantification of flavonoids, which can act as endogenous auxin transport inhibitors, showed a deficiency in the induction of free naringenin, isoliquiritigenin, quercetin, and hesperetin in cre1 roots compared with wild-type roots 24 h after inoculation with rhizobia. Coinoculation of roots with rhizobia and the flavonoids naringenin, isoliquiritigenin, and kaempferol, or with the synthetic auxin transport inhibitor 2,3,5,-triiodobenzoic acid, rescued nodulation efficiency in cre1 mutants and allowed auxin transport control in response to rhizobia. Our results suggest that CRE1-dependent cytokinin signaling leads to nodule initiation through the regulation of flavonoid accumulation required for local alteration of polar auxin transport and subsequent auxin accumulation in cortical cells during the early stages of nodulation.


Protective effect of Hypericum perforatum L. on serum and hair trace elements in rats 7,12-dimethylbenz[a]anthracene-induced oxidative stress.
The study was designed to assess the effect of Hypericum perforatum L. (H.P) on serum and hair trace elements and mineral levels, oral administration of 7,12-dimethylbenz[a]anthracene (DMBA) induced oxidative stress in Sprague-Dawley female rats. Analysis of the trace element has been carried out using atomic absorption spectrophotometer method at end of 60th day. It has been found out that the DMBA group contained statistically lower Zn and Cr compared to the control group (p<0.01) and (p<0.05), Cu, Mg and Na contained higher than control group (p<0.05), (p<0.05) and (p<0.05). In DMBA+H.P group, Zn higher and Na lower than DMBA group (p<0.05), (p<0.05), in hair samples Cd, K and Zn contained lower DMBA compared to the control group (p<0.05), (p<0.05) and (p<0.05). In group DMBA+H.P, Cd was higher than DMBA group and Cr lowered accordance with control group (p<0.05). The present study demonstrated significantly positive and beneficial effect of H.P on the concentration levels of Zn and Na in serum, also on Cd levels in hair between DMBA and DMBA+H.P groups.


Half-Site DNA sequence and spacing length contributions to PrrA binding to PrrA site 2 of RSP3361 in Rhodobacter sphaeroides 2.4.1.
The consensus DNA binding sequence for PrrA, a global regulator in Rhodobacter sphaeroides 2.4.1, is poorly defined. We have performed mutational analysis of PrrA site 2, of the RSP3361 gene, to which PrrA binds in vitro (J. M. Eraso and S. Kaplan, J. Bacteriol. 191:4341-4352, 2009), to further define the consensus sequence for DNA binding. Two half-sites of equal length, containing 6 nucleotides each, were required for PrrA binding to this DNA sequence. Systematic nucleotide substitutions in both inverted half-sites led to a decrease in binding affinity of phosphorylated PrrA in vitro, the level of which was dependent on the substitution. The reduced binding affinities were confirmed by competition experiments and led to proportional decreases in the expression of lacZ transcriptional fusions to the RSP3361 gene in vivo. The 5-nucleotide spacer region between the half-sites was found to be optimal for PrrA binding to the wild-type half-sites, as shown by decreased PrrA DNA binding affinities to synthetic DNA sequences without spacer regions or with spacer regions ranging from 1 to 10 nucleotides. The synthetic spacer region alleles also showed decreased gene expression in vivo when analyzed using lacZ transcriptional fusions. We have studied three additional DNA sequences to which PrrA binds in vitro. They are located in the regulatory regions of genes positively regulated by PrrA and contain spacer regions with 5 or 8 nucleotides. We demonstrate that PrrA can bind in vitro to DNA sequences with different lengths in the spacer regions between the half-sites.


Relative contributions of extracellular Ca2+ and Ca2+ stores to smooth muscle contraction in arteries and arterioles of rat, guinea-pig, dog and rabbit.
1. These studies describe the functional effects of modulation of the sarcoplasmic reticulum (SR) Ca2+ stores at three levels of the vasculature: (i) large arteries (rat and guinea-pig aorta); (ii) small resistance arteries (rat tail artery, rabbit mesenteric artery, dog mesenteric artery); and (iii) arterioles (guinea-pig submucosal arterioles of the small intestine). 2. All tissues responded to phenylephrine (PE; 10 mumol/L) with a transient contraction in Ca(2+)-free Krebs', reflecting Ca2+ release from PE-sensitive Ca2+ stores. After pretreatment with cyclopiazonic acid (CPA; 30 mumol/L) or thapsigargin (TSG; 1 mumol/L), putative SR Ca2+ pump inhibitors, the PE-induced contraction in a Ca(2+)-free medium was significantly inhibited in arterial tissues at all levels of the vasculature. Similarly, ryanodine (RYA; 30 mumol/L), an agonist that enhances Ca2+ release from the SR, also reduced the PE contraction in a Ca(2+)-free solution. 3. CPA or TSG alone in the presence of extracellular Ca2+, caused marked and sustained contraction in the rat and guinea-pig aorta and marked but transient or no contraction in the resistance arteries. In the rat and guinea-pig aorta, RYA caused a slowly developing tension. Little increase in basal tension was produced by RYA in resistance arteries and arterioles. 4. The findings show that an agonist-releasable Ca2+ pool is present at all levels of the vasculature that is independent of the size of the vessels and suggest that under normal physiological conditions there is an intimate balance between the roles of the plasma membrane and of the SR in the maintenance of vascular contractility. It appears that the role of the SR diminishes as the arteries become smaller, while Ca2+ fluxes across the plasma membrane predominates.


Manipulation of prostate cancer metastasis by locus-specific modification of the CRMP4 promoter region using chimeric TALE DNA methyltransferase and demethylase.
Prostate cancer is the most commonly diagnosed non-cutaneous cancer and one of the leading causes of cancer death for North American men. Whereas localized prostate cancer can be cured, there is currently no cure for metastatic prostate cancer. Here we report a novel approach that utilizes designed chimeric transcription activator-like effectors (dTALEs) to control prostate cancer metastasis. Transfection of dTALEs of DNA methyltransferase or demethylase induced artificial, yet active locus-specific CpG and subsequent histone modifications. These manipulations markedly altered expression of endogenous CRMP4, a metastasis suppressor gene. Remarkably, locus-specific CpG demethylation of the CRMP4 promoter in metastatic PC3 cells abolished metastasis, whereas locus-specific CpG methylation of the promoter in non-metastatic 22Rv1 cells induced metastasis. CRMP4-mediated metastasis suppression was found to require activation of Akt/Rac1 signaling and down-regulation of MMP-9 expression. This proof-of-concept study with dTALEs for locus-specific epigenomic manipulation validates the selected CpG methylation of CRMP4 gene as an independent biomarker for diagnosis and prognosis of prostate cancer metastasis and opens up a novel avenue for mechanistic research on cancer biology.


[Application of augmented reality navigation combined with indocyanine green fluorescence imaging technology in the accurate guidance of laparoscopic anatomical segment 8 liver resection].
Objective: To investigate the application value of augmented reality navigation combined with indocyanine green(ICG) fluorescence imaging technology in laparoscopic anatomical segment 8 liver resection. Methods: Clinical and pathological data from 8 patients with hepatocellular carcinoma located in segment 8 of the liver admitted to the First Department of Hepatobiliary Surgery,Zhujiang Hospital,Southern Medical University from October 2021 to October 2022 were collected restrospectively. Among them,there were 5 males and 3 females,aged between 40 and 72 years. During the operation,the self-developed laparoscopic augmented reality surgical navigation system was used to integrate the three-dimensional liver model with the laparoscopic scene,and ICG fluorescence imaging technology was used to guide the anatomical liver resection of segment 8. The predicted liver resection volume and actual liver resection volume,related surgical indicators and postoperative complications were analyzed. Results: Among the 8 patients, 4 underwent laparoscopic anatomical segment 8 liver resection,1 underwent laparoscopic anatomical ventral subsegment of segment 8 liver resection,2 underwent laparoscopic anatomical ventral subsegment combined with medial subsegment of segment 8 liver resection, and 1 underwent laparoscopic anatomical dorsal subsegment of segment 8 liver resection. All operations were completed under the guidance of augmented reality navigation combined with ICG fluorescence imaging,without conversion to open surgery. The operation time was (276.3±54.8)minutes(range:200 to 360 minutes). Intraoperative blood loss was (75.0±35.4)ml(range:50 to 150 ml). No blood transfusion was performed during the operation. The length of postoperative hospital stay was (7.6±0.8)days(range:7 to 9 days). There were no deaths or postoperative complications such as bleeding or biliary fistula during the perioperative period. Conclusion: Augmented reality navigation combined with ICG fluorescence imaging technology can guide the implementation of laparoscopic anatomical segment 8 liver resection.


Heat shock protein 90 indirectly regulates ERK activity by affecting Raf protein metabolism.
Extracellular signal-regulated protein kinase (ERK) has been implicated in the pathogenesis of several nerve system diseases. As more and more kinases have been discovered to be the client proteins of the molecular chaperone Hsp90, the use of Hsp90 inhibitors to reduce abnormal kinase activity is a new treatment strategy for nerve system diseases. This study investigated the regulation of the ERK pathway by Hsp90. We showed that Hsp90 inhibitors reduce ERK phosphorylation without affecting the total ERK protein level. Further investigation showed that Raf, the upstream kinase in the Ras-Raf-MEK-ERK pathway, forms a complex with Hsp90 and Hsp70. Treating cells with Hsp90 inhibitors facilitates Raf degradation, thereby down-regulating the activity of ERK.


Intra-operative cell salvage in urological surgery: a systematic review and meta-analysis of comparative studies.
To evaluate systematically the safety and efficacy of intra-operative cell salvage (ICS) in urology. A search of Medline, Embase and Cochrane Library to August 2017 was performed using methods pre-published on PROSPERO. Reporting followed the Preferred Reporting Items for Systematic Review and Meta-analysis guidelines. Eligible titles were comparative studies published in English that used ICS in urology. Primary outcomes were allogeneic transfusion rates (ATRs) and tumour recurrence. Secondary outcomes were complications and cost. Fourteen observational studies were identified, with a total of 4 536 patients. ICS was compared with no the blood conservation technique (seven studies), preoperative autologous donation (PAD; five studies) or both (two studies). Cohorts underwent open prostatectomy (11 studies), open cystectomy (two studies) or open partial nephrectomy (one study). Meta-analysis was possible only for ATRs within prostatectomy studies. In this setting, ICS reduced ATR compared with no the blood conservation technique (odds ratio [OR] 0.34, 95% confidence interval [CI] 0.15-0.76) but not PAD (OR 0.76, 95% CI 0.39-1.31). In the non-prostatectomy setting, ATRs amongst patients who underwent ICS were significantly higher or similar in one and two studies, respectively. Tumour recurrence was found to be significantly less common (two studies), similar (eight studies) or not measured (four studies). All six studies reporting complications found no difference in their ICS cohorts. Regarding cost, one study from 1995 found ICS more expensive than PAD, while two more recent studies found ICS to be cheaper than no blood conservation technique. As a result of inter-study heterogeneity, meta-analyses were not possible for recurrence, complications or cost. Low-level evidence exists that, compared with other blood conservation techniques, ICS reduces ATR and cost while not affecting complications. It does not appear to increase tumour recurrence post-prostatectomy, although follow-up durations were short. Small study sizes and short follow-ups mean conclusions cannot be drawn with regard to recurrence after nephrectomy or cystectomy. Randomized trials with long-term follow-up evaluating ICS in urology are required.


Lymphocyte-platelet crosstalk in Graves' disease.
Platelets can modulate lymphocytes' role in the pathophysiology of thyroid autoimmune diseases. The present study was performed to clarify the status of platelet-lymphocyte subpopulations aggregation in circulating blood in patients with Graves' disease (GD). One hundred and fifty patients with GD (GD group) and 45 hyperthyroid patients with toxic multinodular goiter (TMG group) were recruited in the study. Control group consisted 150 healthy subjects. Immunophenotyping of lymphocytes was performed by flow cytometry. Detection of lymphocyte-platelet aggregates (LPAs) was done using light microscope after Ficoll-gradient centrifugation. The group of GD patients exhibited reduced CD8 lymphocyte and higher CD19 cell counts compared with TMG group and healthy controls. A greater number of activated CD3, HLA-DR+ lymphocytes were observed in GD than in TMG group and control group. GD group was characterized by lower blood platelet count (232 ± 89 × 10 cells/µL) than TMG group (251 ± 97 × 10 cells/µL; P < 0.05) and control group (262 ± 95 × 10 cells/µL; P < 0.05). In GD group, more platelet-bound lymphocytes (332 ± 91 /µL) were found than that in TMG group (116 ± 67/µL, P < 0.005) and control group (104 ± 58 /µL; P < 0.001). GD is associated with higher levels of activated lymphocytes and lymphocyte-platelet aggregates.


Shortage of professional nursing practice: a reframing of the shortage problem.
The nursing shortage is discussed in terms of supply, demand, and wages, and the current situation is compared with previous nursing shortages. Reasons for the shortage are examined and reframing the problem from a shortage of nurses to a shortage of professional nursing practice is suggested as a strategic first step in solving the problems of nurse staffing in acute care general hospitals. Four kinds of shortage situations are identified; only one kind, vacancy, results from vacant positions. The other three types, transient, position, and scheduling, are the result of administrative decisions regarding the use of nursing personnel. A fundamental restructuring of how hospitals use nurses is needed, and this solution will in the long run be more effective in solving the shortage problem than is the effort simply to add more nurses into the existing system.


Activators of protein kinase C inhibit sodium transport in A6 epithelia.
To evaluate the role of protein kinase C in sodium transport via amiloride-sensitive sodium channels, we studied the effect of activators of protein kinase C on short-circuit current in epithelia formed by A6 cells in culture. In A6 epithelia, short-circuit current is equivalent to net sodium transport and is blocked by low concentrations of amiloride added to the solution bathing the apical surface. After any of four different activators of protein kinase C [phorbol 12,13-dibutyrate (20 ng/ml), phorbol 12-myristate 13-acetate (20 ng/ml), 1-oleoyl-2-acetylglycerol (50 micrograms/ml), and mezerein (10 ng/ml)] was added to the solution bathing the apical surface, short-circuit current fell, and electrical resistance rose. Nystatin added to the apical solution reversed the inhibition of short-circuit current, indicating that activators of protein kinase C inhibited transport at amiloride-sensitive sodium channels in the apical plasma membrane. Under some conditions, the activators also stimulated amiloride-insensitive short-circuit current. The ion transport represented by the amiloride-insensitive short-circuit current appears to be the result of basal to apical transport of chloride, but this has not been established conclusively.


A subclass of serum anti-ZnT8 antibodies directed to the surface of live pancreatic β-cells.
The islet-specific zinc transporter ZnT8 is a major self-antigen found in insulin granules of pancreatic β-cells. Frequent insulin secretion exposes ZnT8 to the cell surface, but the humoral antigenicity of the surface-displayed ZnT8 remains unknown. Here we show that a membrane-embedded human ZnT8 antigen triggered a vigorous immune response in ZnT8 knock-out mice. Approximately 50% of serum immunoreactivities toward ZnT8 were mapped to its transmembrane domain that is accessible to extracellular ZnT8 antibody (ZnT8A). ZnT8A binding was detected on live rat insulinoma INS-1E cells, and the binding specificity was validated by a CRISPR/Cas9 mediated ZnT8 knock-out. Applying established ZnT8A assays to purified serum antibodies from patients with type 1 diabetes, we detected human ZnT8A bound to live INS-1E cells, whereas a ZnT8 knock-out specifically reduced the surface binding. Our results demonstrate that ZnT8 is a cell surface self-antigen, raising the possibility of a direct involvement in antibody-mediated β-cell dysfunction and cytotoxicity.


Glycoprotein H and α4β1 integrins determine the entry pathway of alphaherpesviruses.
Herpesviruses enter cells either by direct fusion at the plasma membrane or from within endosomes, depending on the cell type and receptor(s). We investigated two closely related herpesviruses of horses, equine herpesvirus type 1 (EHV-1) and EHV-4, for which the cellular and viral determinants routing virus entry are unknown. We show that EHV-1 enters equine epithelial cells via direct fusion at the plasma membrane, while EHV-4 does so via an endocytic pathway, which is dependent on dynamin II, cholesterol, caveolin 1, and tyrosine kinase activity. Exchange of glycoprotein H (gH) between EHV-1 and EHV-4 resulted in rerouting of EHV-1 to the endocytic pathway, as did blocking of α4β1 integrins on the cell surface. Furthermore, a point mutation in the SDI integrin-binding motif of EHV-1 gH also directed EHV-1 to the endocytic pathway. Cumulatively, we show that viral gH and cellular α4β1 integrins are important determinants in the choice of alphaherpesvirus cellular entry pathways.


Feasibility and accuracy of a new mobile electrocardiography device, ER-2000(®), in the diagnosis of arrhythmia.
We performed this study to evaluate the feasibility and accuracy of a new mobile electrocardiography (ECG) device, ER-2000(®), in detecting cardiac arrhythmia, by comparing it to a 12-lead ECG used as the gold standard. Mode 1 of ER-2000(®) was recorded using three electrodes with cables attached to the anterior chest wall, and mode 2 was recorded using the side chest channel and finger channel. Standard 12-lead ECG was used to record with a speed of 25 mm/s, simultaneously. Seventeen patients with complaints of palpitation were enrolled. Twelve-lead ECG revealed normal sinus rhythm in three patients, sinus tachycardia in one, atrial fibrillation (AF) in two, atrial tachycardia (AT) in one, first degree atrioventricular block in one, pacing rhythm in two patients who underwent permanent pacemaker implantation, AF with intermittent ventricular pacing in one, complete right bundle branch block in one, J-wave elevation in one, narrow QRS tachycardia in one, atrial premature beat (APB) in one, ventricular premature beat (VPB) in one, and narrow QRS tachycardia with intermittent aberrant conduction in one. Rhythm diagnosis obtained by the two different modes of ER-2000(®) was correlated with that obtained by the 12-lead ECG in all patients except in one in whom ER-2000(®) showed one APB while 12-lead ECG showed sinus rhythm. R-R interval was accurately recorded despite the detailed morphology of QRS, and T-wave was somewhat modified with the use of ER-2000(®). A pacing blip detected by 12-lead ECG was not detected by ER-2000(®) despite a similar wide QRS duration in the paced QRS. A rhythm strip obtained using ER-2000(®) is accurate in diagnosing arrhythmia, despite some differences in the detailed morphology of the QRS and T-wave, and the pacing spike compared to those obtained by the 12-lead ECG.


[Assessing quality in clinical educational setting: Italian validation of the clinical learning environment and supervision (CLES) scale].
The clinical learning in the wards is one of the most important moment in nursing education: Instruments assessing the quality of the clinical settings as perceived by students may inform and support the best didactic and tutorial choices. In the last 10 years, three clinical learning environment assessment tools for Bachelor Nursing students has been validated. The aim of this study was to validate in the Italian context the Clinical Learning Environment and Supervision scale (CLES) developed in Finland and considered the gold standard within the instruments available. CLES's face and content validity has been evaluated with forward and backward translation and involving a selected group of experts in the field. 117 Bachelor Nursing Students from Udine and Verona University have been enrolled in the study. 28 out of 117 students have filed the instrument twice in order to measure test-retest reliability. The internal consistency has been evaluated with Cronbach alpha. Kurtosis and skewness was measured in order to verify the answers variability distribution. The CLES Cronbach' alpha was 0.96 and the test-retest reliability was r = -0.89. The validity and reliability of the scale has been confirmed. Further research is needed to define a cut-off in order to recognize positive clinical learning environments. The CLES practical use could support in the process of the teaching hospital accreditation and in the evaluation of the effectiveness of the tutorial strategies activated in the wards.


Heparin, heparan sulfate and heparanase in inflammatory reactions.
Heparan sulfate (HS) proteoglycans at the cell surface and in the extracellular matrix of most animal tissues are essential in development and homeostasis, and are implicated in disease processes. Emerging evidence demonstrates the important roles of HS in inflammatory reactions, particularly in the regulation of leukocyte extravasation. Heparin, a classical anticoagulant, exhibits anti-inflammatory effects in animal models and in the clinic, presumably through interference with the functions of HS, as both polysaccharides share a high similarity in molecular structure. Apart of regulation during biosynthesis, the structures of HS and heparin are significantly modulated by heparanase, an endoglycosidase that is upregulated in a number of inflammatory conditions. Exploring the physiological roles of HS and heparin and the mode of heparanase action in modulating their functions during inflammation responses is of importance for future studies.


Unusual Long Survival with a Giant Invasive Pheochromocytoma of an Incompatible Patient.
Pheochromocytomas (PHEOs) are rare neuroendocrine tumors and about 2-13% of PHEOs are malignant. Predicting malignancy in PHEO cases with invasion but without metastasis is still controversial in the literature. This study presents an unusual long survival with a giant invasive PHEO in an incompatible patient and a review of the literature. In 1989, a 23-year-old female patient was operated for a giant adrenal mass with a pathological final diagnosis of PHEO. Information to the patient's family was provided about the short life span of the patient in the postoperative period because the tumor could not be totally resected. The patient started using regular antihypertensive drugs only after 1994. In 1994, 3700 mBq 131-I-metaiodobenzylguanidine (MIBG) treatment was given. Since then, no specific treatment was administered for PHEO due to patient incompatibility. She was diagnosed with type 2 diabetes mellitus at the age of 40 years and had a cerebrovascular accident due to hypertension at the age of 42. New abdominal computed tomography (CT) showed a right-sided 75 x 37 mm irregular and heterogeneous mass lesion extending inferiorly from the diaphragmatic crus level located in the right adrenal locus compatible with local recurrence. There was no I-123-MIBG uptake. She refused to have advanced workup and further treatment options. Malignant PHEOs reduce overall survival as a consequence of excessive catecholamine release, large tumor burden, and malignancy-related complications. Currently, the treatment of a malignant PHEO still has difficulties for both patients and doctors. Main treatment options for malignant PHEOs are primarily surgical excision. The effect of radionuclide therapy on survival time still remains to be determined. Efforts should be made to identify clinical, biochemical, and pathological criteria for malignancy and to develop new therapies in these patients with malignancy. The clinical course of malignant PHEOs is remarkably variable. Disease-specific survival rate changes from 58 to 88.1% at five years in the literature. Recent discoveries have enhanced new options for treatment, from radionuclide therapy and targeted molecular therapy to immunotherapy. A multidisciplinary approach is needed to individualize treatment in patients with malignant and invasive PHEO.


Prednisone-free maintenance immunosuppression-a 5-year experience.
Concern persists that prednisone-free maintenance immunosuppression in kidney transplant recipients will be associated with an increase in late allograft dysfunction and graft loss. We herein report 5-year follow-up of a trial of prednisone-free maintenance immunosuppression. From October 1, 1999, through January 31, 2005, at our center, 589 kidney transplant recipients were treated with a protocol incorporating discontinuation of their prednisone on postoperative day 6. At 5 years, actuarial patient survival was 91%; graft survival, 84%; death-censored graft survival, 92%; acute rejection-free graft survival, 84% and chronic rejection-free graft survival, 87%. The mean serum creatinine level (+/-SD) at 1 year was 1.6 +/- 0.6; at 5 years, 1.7 +/- 0.8. In all, 86% of kidney recipients with functioning grafts remain prednisone-free as of April 30, 2005. As compared with historical controls, recipients on prednisone-free maintenance immunosuppression had a significantly lower rate of a number of complications, including cataracts (p < 0.001), posttransplant diabetes mellitus (p < 0.001), avascular necrosis (p = 0.001), and fractures (p = 0.004). We conclude that prednisone-related side effects can be minimized in a protocol incorporating prednisone-free maintenance immunosuppression. Five-year graft outcome remains good.


A quantitative approach for analyzing the spatio-temporal distribution of 3D intracellular events in fluorescence microscopy.
Analysis of the spatial distribution of endomembrane trafficking is fundamental to understand the mechanisms controlling cellular dynamics, cell homeostasy, and cell interaction with its external environment in normal and pathological situations. We present a semi-parametric framework to quantitatively analyze and visualize the spatio-temporal distribution of intracellular events from different conditions. From the spatial coordinates of intracellular features such as segmented subcellular structures or vesicle trajectories, QuantEv automatically estimates weighted densities that are easy to interpret and performs a comprehensive statistical analysis from distribution distances. We apply this approach to study the spatio-temporal distribution of moving Rab6 fluorescently labeled membranes with respect to their direction of movement in crossbow- and disk-shaped cells. We also investigate the position of the generating hub of Rab11-positive membranes and the effect of actin disruption on Rab11 trafficking in coordination with cell shape.


Geologic evidence for recurrent moderate to large earthquakes near charleston, South Carolina.
Multiple generations of earthquake-induced sand blows in Quaternary sediments and soils near Charleston, South Carolina, are evidence of recurrent moderate to large earthquakes in that area. The large 1886 earthquake, the only historic earthquake known to have produced sand blows at Charleston, probably caused the youngest observed blows. Older (late Quaternary) sand blows in the Charleston area indicate at least two prehistoric earthquakes with shaking severities comparable to the 1886 event.


Working better together: new approaches for understanding the value and challenges of organizational partnerships.
Inter-agency partnerships are critical for addressing the interrelated circumstances associated with the social and health determinants of health inequalities. However, there are many challenges in evaluating partnership processes and outcomes. We discuss a mixed methods study that explored partnership processes in an innovative program that aims to promote social and economic inclusion for young newly arrived refugees. A theoretically informed evaluation was designed and data collected in three ways: an organizational ethnographic approach; a partnership self-assessment tool and semi-structured interviews. Partnership assessments and interviews were collected at two points in time providing progressive process data. Analyses explore divergent levels of staff satisfaction with the partnership's operations, particularly between staff working in program development (strategic management) and program delivery (service provision) roles. Follow-up data collection indicated satisfaction with partnership processes had improved. The partnership did achieve its aim of increasing the level of cooperation between service providers to support young people from refugee backgrounds. This paper presents insights into how to evaluate inter-agency partnerships and reports both methodological and empirical findings. It provides an approach for a better understanding of the levels at which individuals operate within such partnerships, indicates areas where support and attention is needed.


The organizer in evolution-gastrulation and organizer gene expression highlight the importance of Brachyury during development of the coral, Acropora millepora.
Organizer activity, once thought to be restricted to vertebrates, has ancient origins. However, among non-bilaterians, it has only been subjected to detailed investigation during embryonic development of the sea anemone, Nematostella vectensis. As a step toward establishing the extent to which findings in Nematostella can be generalized across the large and diverse phylum Cnidaria, we examined the expression of some key organizer and gastrulation genes during the embryonic development of the coral Acropora millepora. Although anemones and corals both belong to the cnidarian class Anthozoa, the two lineages diverged during the Cambrian and the morphological development of Acropora differs in several important respects from that of Nematostella. While the expression patterns of the key genes brachyury, bmp2/4, chordin, goosecoid and forkhead are broadly similar, developmental differences between the two species enable novel observations, and new interpretations of their significance. Specifically, brachyury expression during the flattened prawnchip stage before gastrulation, a developmental peculiarity of Acropora, leads us to suggest that it is the key gene demarcating ectoderm from endoderm in Acropora, and by implication in other cnidarians, whereas previous studies in Nematostella proposed that forkhead plays this role. Other novel observations include the transient expression of Acropora forkhead in scattered ectodermal cells shortly after gastrulation, and in the developing mesenterial filaments, with no corresponding expression reported in Nematostella. In addition, the expression patterns of goosecoid and bmp2/4 confirm the fundamental bilaterality of the Anthozoa.


Water activity effects on geranyl acetate synthesis catalyzed by novozym in supercritical ethane and in supercritical carbon dioxide.
The esterification reaction of geraniol with acetic acid catalyzed by Novozym was studied in supercritical ethane (sc-ethane) and in supercritical carbon dioxide (sc-CO(2)). Water activity (a(W)) had a very strong effect on enzyme activity, with reaction rates increasing up to a(W) = 0.25 and then decreasing for higher a(W). Salt hydrate pairs could not prevent changes in a(W) during the course of reaction but were able to control a(W) to some extent and had a beneficial effect on both initial rates of esterification and conversion in sc-ethane. The enzyme was more active in sc-ethane than in sc-CO(2), confirming the deleterious effect of the latter already observed with some enzymes. Temperatures between 40 and 60 degrees C did not have a strong effect on initial rates of esterification, although reaction progress declined considerably in that temperature range. For the mixture of 50 mM acetic acid plus 200 mM geraniol, 100% conversion was achieved at a reaction time of 10 h at 40 degrees C, 100 bar, an a(W) of incubation of 0.25, and a Novozym concentration of 0.55 mg cm(-)(3) in sc-ethane. Conversion was below 50% in sc-CO(2) at otherwise identical conditions. With an equimolar mixture of the two substrates (100 mM), 98% conversion was reached at 10 h of reaction in sc-ethane (73% conversion in sc-CO(2)).


Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.).
Genes involved in flavonoid and stilbene biosynthesis were isolated from grape (Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by screening a cDNA library, obtained from mRNA from seedlings grown in light for 48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA clone coding for stilbene synthase (StSy) was isolated by probing the library with a specific oligonucleotide. These clones were sequenced and when the putative products were compared to the published amino acid sequence for corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to 90% (CHS and PAL). The analysis of the genomic organization and expression of these genes in response to light shows that PAL and StSy genes belong to large multigene families, while the others are present in one to four copies per haploid genome. The steady-state level of mRNAs encoded by the flavonoid biosynthetic genes as determined in young seedlings is coordinately induced by light, except for PAL and StSy, which appear to be constitutively expressed.


Cardiac mitochondrial calcium loading capacity is severely affected after chronic cholestasis in Wistar rats.
Cardiovascular changes correlated with some forms of hepatic disease are being reported in the literature. The aim of this work was to characterize cardiac mitochondrial bioenergetics and calcium buffering capacity in Wistar rats injected with six weekly doses of alpha-naphthylisothiocyanate (ANIT), a compound known to induce cholestasis in animal models. Isolated heart mitochondria were obtained from both injected and control animals and bioenergetic parameters were measured, as well as the capacity to buffer externally added calcium and the mitochondrial content of reduced protein thiol groups. Blood biochemistry analyses were obtained at the initial and end points of treatment. The in vitro ANIT effect on isolated heart mitochondria was also studied. Our results showed that the respiratory control ratio was the only parameter affected in injected animals (p < .05, n = 5). Nevertheless, heart mitochondria from injected animals showed an inability to accumulate added calcium owing to an increased susceptibility to the calcium-dependent mitochondrial permeability transition (p < .0001, n = 5). The effects were still present 1 week after ending ANIT administration, when serum markers for liver injury and hyperbilirubinemia were already abated (although in the presence of bile duct proliferation). To our knowledge, this is the first time that cardiac mitochondrial calcium homeostasis and mitochondrial respiratory ratio are seen affected during ANIT-induced cholestasis, prevailing even in the absence of hepatic damage serum markers.


Protein kinases in Toxoplasma gondii.
Toxoplasma gondii is an obligatory intracellular pathogen that causes life threatening illness in immunodeficient individuals, miscarriage in pregnant woman, and blindness in newborn children. Similar to any other eukaryotic cell, protein kinases play critical and essential roles in the Toxoplasma life cycle. Accordingly, many studies have focused on identifying and defining the mechanism of function of these signalling proteins with a long-term goal to develop anti-Toxoplasma therapeutics. In this review, we briefly discuss classification and key components of the catalytic domain which are critical for functioning of kinases, with a focus on domains, families, and groups of kinases within Toxoplasma. More importantly, this article provides a comprehensive, current overview of research on kinase groups in Toxoplasma including the established eukaryotic AGC, CAMK, CK1, CMGC, STE, TKL families and the apicomplexan-specific FIKK, ROPK and WNG family of kinases. This work provides an overview and discusses current knowledge on Toxoplasma kinases including their localization, function, signalling network and role in acute and chronic pathogenesis, with a view towards the future in probing kinases as viable drug targets.


Centromeric distribution of 350-family in Dasypyrum villosum and its application to identifying Dasypyrum chromatin in the wheat genome.
Variants of the 350-family sequences were isolated from Dasypyrum villosum genomic DNA. Although the consensus sequence shared 86% homology to p380 and 84% homology to pHvNAU62 of D. villosum, a 63 bp long region including variable microsatellite repeat (TG)(7-9) was found to be absent in pHvNAU62 or much different from p380. This sequence also shared 67% homology to the 350-family of rye. The representative clone pDvTU383 was localized on the D. villosum chromosomes by using the sequential C-banding and fluorescence in situ hybridization (FISH). Combining with the probes of pTa71 and pTa794 containing 18S-5.8S-28S rDNA (NOR) and 5S rDNA multigene families, respectively, the pDvTU383 probe allowed identification of all seven pairs of D. villosum chromosomes. The pair of the 1V chromosomes fluoresced strong in situ hybridization signals of 18S-5.8S-28S rRNA genes in the NORs of 1VS, and the pair of the 5V chromosomes had weak signals of 5S rRNA genes at the end of 5VS. The pair of the 4V chromosomes showed sub-telomeric signals of pDvTU383 probe on the both arms, however, the signal of the short arm is weaker than that of the long arm. The pair of the 7V chromosomes showed no signals of pDvTU383 on the both sub-telomeric regions. The pair of the 6V chromosomes showed a characteristic interstitial signal of pDvTU383 on the short arm. The pairs of the 2V and 3V chromosomes were distinguished by the difference of interstitial signals of pDvTU383 on their long arms. Furthermore, hybridization signals of pDvTU383 were also visualized on centromeres of all chromosomes. Based on the above D. villosum chromosomal patterns made by FISH, one pair of the 4V chromosomes originated from D. villosum were found in a trigeneric hybrid line involving wheat, Dasypyrum and Thinopyrum. The sequence pDvTU383 provides an effective probe for further analysis of the D. villosum genome.