Levels of service user satisfaction in secure settings--a survey of the association between perceived social climate, perceived therapeutic relationship and satisfaction with forensic services.
The interests of users should lead service developments. However, it has been claimed that forensic mental health services have largely ignored examining users' views on the nature and quality of the service offered to them. Perceived social climate and perceived therapeutic relationship are viewed as important indicators of treatment outcome; however previous findings about how these variables may be associated with satisfaction with forensic services are equivocal. This study aims to assess the levels of service user satisfaction in forensic in-patient settings in one mental health trust and explore how the perceived therapeutic relationship of the service users with their key-workers and the perceived social climate of the wards are associated with service user satisfaction. A cross-sectional survey design was chosen and an independent researcher facilitated the completion of three standardised assessments measuring service user satisfaction, therapeutic relationships and the social climate of the ward. Forty-four service users detained in secure settings completed the standardised assessments. The study was conducted in four medium secure and three low secure units in one NHS mental health trust. The data was analysed to examine the level of satisfaction with services and how both the therapeutic relationship and the ward environment were related to levels of satisfaction. The majority of service users who responded were generally satisfied with services; "rehabilitation" and "perceived safety" were viewed most positively. Service users' perceptions about the social climate of the ward were found to have a significant relationship with service users' satisfaction with forensic services. However, the variables with the strongest association with satisfaction with forensic services are service users' perceptions about the nature of therapeutic relations with staff. This study indicates that service users' satisfaction with forensic services is strongly associated with their experiences of the therapeutic relationship with their key-workers and the social climate of the ward. The findings emphasize the importance of forming and maintaining effective therapeutic relations and reinforce the need to maintain a therapeutic environment free of aggressive tension and threats of violence. The results also highlight the potential for service users to be dissatisfied with their financial situation following admission.


Symptomatic response to divalproex in subtypes of conduct disorder.
To investigate response to Divalproex sodium (DVPX) with respect to Reactive/Affective/Defensive/Impulsive (RADI) and Proactive/Instrumental/Premeditated (PIP) aggression among adolescent males with conduct disorder (CD), using results from a randomized, double-blind, placebo-controlled trial. It was hypothesized that DVPX response among participants with RADI aggression would be greater than among those with PIP aggression. Fifty-eight ethnically diverse males with severe CD were assigned to High Distress (HDCD) or Low Distress (LDCD) Conduct Disorder, corresponding with RADI and PIP aggression, respectively. Following a 1-week washout, all subjects were randomized to a high dose (up to 1,500 mg/day) or low dose (up to 250 mg/day) of DVPX. Baseline and endpoint assessments included Clinical Global Impression (CGI), Achenbach Self Report (YSR), and Weinberger Adjustment Inventory (WAI-62). Response to DVPX was significantly higher in the HDCD group (64%) than in the LDCD group (22%) in the high-dose treatment group (p = 0.03). Mean weekly WAI-62 distress scores declined significantly among the HDCD subjects than among LDCD subjects in the high-dose group. These results support the utility of mood stabilizing agents such as DVPX in treating patients with disorders characterized by the RADI pattern of aggression, including those with severe CD.


Disruption of Glucagon-Like Peptide 1 Signaling in Sim1 Neurons Reduces Physiological and Behavioral Reactivity to Acute and Chronic Stress.
Organismal stress initiates a tightly orchestrated set of responses involving complex physiological and neurocognitive systems. Here, we present evidence for glucagon-like peptide 1 (GLP-1)-mediated paraventricular hypothalamic circuit coordinating the global stress response. The GLP-1 receptor (Glp1r) in mice was knocked down in neurons expressing single-minded 1, a transcription factor abundantly expressed in the paraventricular nucleus (PVN) of the hypothalamus. Mice with single-minded 1-mediated Glp1r knockdown had reduced hypothalamic-pituitary-adrenal axis responses to both acute and chronic stress and were protected against weight loss associated with chronic stress. In addition, regional Glp1r knockdown attenuated stress-induced cardiovascular responses accompanied by decreased sympathetic drive to the heart. Finally, Glp1r knockdown reduced anxiety-like behavior, implicating PVN GLP-1 signaling in behavioral stress reactivity. Collectively, these findings support a circuit whereby brainstem GLP-1 activates PVN signaling to mount an appropriate whole-organism response to stress. These results raise the possibility that dysfunction of this system may contribute to stress-related pathologies, and thereby provide a novel target for intervention. Dysfunctional stress responses are linked to a number of somatic and psychiatric diseases, emphasizing the importance of precise neuronal control of effector pathways. Pharmacological evidence suggests a role for glucagon-like peptide-1 (GLP-1) in modulating stress responses. Using a targeted knockdown of the GLP-1 receptor in the single-minded 1 neurons, we show dependence of paraventricular nucleus GLP-1 signaling in the coordination of neuroendocrine, autonomic, and behavioral responses to acute and chronic stress. To our knowledge, this is the first direct demonstration of an obligate brainstem-to-hypothalamus circuit orchestrating general stress excitation across multiple effector systems. These findings provide novel information regarding signaling pathways coordinating central control of whole-body stress reactivity.


Determinants of gating polarity of a connexin 32 hemichannel.
There is good evidence supporting the view that the transjunctional voltage sensor (V(j)-sensor) of Cx32 and other Group 1 connexins is contained within a segment of the N-terminus that contributes to the formation of the channel pore. We have shown that the addition of negatively charged amino acid residues at several positions within the first 10 amino acid residues reverses the polarity of V(j)-gating and proposed that channel closure is initiated by the inward movement of this region. Here, we report that positive charge substitutions of the 2nd, 5th, and 8th residues maintain the negative polarity of V(j)-gating. These data are consistent with the original gating model. Surprisingly, some channels containing combinations of positive and/or negative charges at the 2nd and 5th positions display bipolar V(j)-gating. The appearance of bipolar gating does not correlate with relative orientation of charges at this position. However, the voltage sensitivity of bipolar channels correlates with the sign of the charge at the 2nd residue, suggesting that charges at this position may have a larger role in determining gating polarity. Taken together with previous findings, the results suggest that the polarity V(j)-gating is not determined by the sign of the charge lying closest to the cytoplasmic entry of the channel, nor is it likely to result from the reorientation of an electrical dipole contained in the N-terminus. We further explore the mechanism of polarity determination by utilizing the one-dimensional Poisson-Nernst-Plank model to determine the voltage profile of simple model channels containing regions of permanent charge within the channel pore. These considerations demonstrate how local variations in the electric field may influence the polarity and sensitivity of V(j)-gating but are unlikely to account for the appearance of bipolar V(j)-gating.


Effectiveness of public deliberation methods for gathering input on issues in healthcare: Results from a randomized trial.
Public deliberation elicits informed perspectives on complex issues that are values-laden and lack technical solutions. This Deliberative Methods Demonstration examined the effectiveness of public deliberation for obtaining informed public input regarding the role of medical evidence in U.S. healthcare. We conducted a 5-arm randomized controlled trial, assigning participants to one of four deliberative methods or to a reading materials only (RMO) control group. The four deliberative methods reflected important differences in implementation, including length of the deliberative process and mode of interaction. The project convened 76 groups between August and November 2012 in four U.S. Chicago, IL; Sacramento, CA; Silver Spring, MD; and Durham, NC, capturing a sociodemographically diverse sample with specific attention to ensuring inclusion of Hispanic, African-American, and elderly participants. Of 1774 people recruited, 75% participated: 961 took part in a deliberative method and 377 participants comprised the RMO control group. To assess effectiveness of the deliberative methods overall and of individual methods, we evaluated whether mean pre-post changes on a knowledge and attitude survey were statistically different from the RMO control using ANCOVA. In addition, we calculated mean scores capturing participant views of the impact and value of deliberation. Participating in deliberation increased participants' knowledge of evidence and comparative effectiveness research and shifted participants' attitudes regarding the role of evidence in decision-making. When comparing each deliberative method to the RMO control group, all four deliberative methods resulted in statistically significant change on at least one knowledge or attitude measure. These findings were underscored by self-reports that the experience affected participants' opinions. Public deliberation offers unique potential for those seeking informed input on complex, values-laden topics affecting broad public constituencies.


Hydrogen breath test and intestinal gas production.
Despite extensive use in clinical practice, difficulties regarding interpretation of hydrogen breath test are still very frequent, even on research grounds. After the administration of a non-absorbable sugar, such as lactulose, an increase of breath hydrogen and methane is evident; this phenomenon is considered an index of colonic fermentation. It is not clear, however, if the levels of these compounds correlate with the presence and severity of functional symptoms, nor if they accurately reflect gas production at colonic level. So far, apart from flatulence, we have no indications regarding the ability of hydrogen or methane to act as biomarkers of intraluminal events. On the other hand, it has been shown that in functional bowel disease a colonic dysbiosis exists, and that the modification of bacterial flora might result in a reduction of symptom severity. Consequently, it is not clear if hydrogen and methane colonic production could have a role in the pathophysiology of functional complaints, but it is possible that other fermentation products should be taken into consideration, such as acetate, propionate, and alcohol.


A shift towards early-age desexing of cats under veterinary care in Australia.
The global problem of unowned domestic cats, driven by their phenomenal reproductive success, carries significant economic, animal welfare and biodiversity costs. Desexing owned cats prior to puberty prevents unwanted litters that contribute to unowned cat populations. The prevalence and predictors of desexing, and the age at which surgery was carried out were investigated using anonymized electronic patient records in the VetCompass Australia database of cats presented to veterinary practices. Of 52,941 cats born between 2010 and 2017, 83.6% were desexed. Among 7463 desexed females, 21.5% had been desexed by 4 months of age, 59.8% by 6 months and 85.4% by 1 year. Sex, breed, location and socioeconomic indices significantly influenced desexing status and age at surgery. Cats born between 2010 and 2017 had greater odds of being desexed than cats born between 1995and 2009 at each age cut-off (≤ 4 months [OR 1.76, CI95 1.58-1.97], ≤ 6 months [OR 1.50, CI95 1.38-1.62] and ≤ 1 year [OR 2.33, CI95 2.11-2.57] p < 0.001). Most cats presented to veterinarians in Australia are desexed. Compared with cats born before 2010, cats born later are significantly younger at desexing but, even so, many cats would have reached sexual maturity before surgery. These findings will inform the design of front-line strategies promoting prepubertal desexing and they demonstrate, for the first time, a shift towards desexing younger cats.


Ca²+-dependent phosphorylation of RyR2 can uncouple channel gating from direct cytosolic Ca²+ regulation.
Phosphorylation of the cardiac ryanodine receptor (RyR2) is thought to be important not only for normal cardiac excitation-contraction coupling but also in exacerbating abnormalities in Ca²+ homeostasis in heart failure. Linking phosphorylation to specific changes in the single-channel function of RyR2 has proved very difficult, yielding much controversy within the field. We therefore investigated the mechanistic changes that take place at the single-channel level after phosphorylating RyR2 and, in particular, the idea that PKA-dependent phosphorylation increases RyR2 sensitivity to cytosolic Ca²+. We show that hyperphosphorylation by exogenous PKA increases open probability (P(o)) but, crucially, RyR2 becomes uncoupled from the influence of cytosolic Ca²+; lowering [Ca²+] to subactivating levels no longer closes the channels. Phosphatase (PP1) treatment reverses these gating changes, returning the channels to a Ca²+-sensitive mode of gating. We additionally found that cytosolic incubation with Mg²+/ATP in the absence of exogenously added kinase could phosphorylate RyR2 in approximately 50% of channels, thereby indicating that an endogenous kinase incorporates into the bilayer together with RyR2. Channels activated by the endogenous kinase exhibited identical changes in gating behavior to those activated by exogenous PKA, including uncoupling from the influence of cytosolic Ca²+. We show that the endogenous kinase is both Ca²+-dependent and sensitive to inhibitors of PKC. Moreover, the Ca²+-dependent, endogenous kinase-induced changes in RyR2 gating do not appear to be related to phosphorylation of serine-2809. Further work is required to investigate the identity and physiological role of this Ca²+-dependent endogenous kinase that can uncouple RyR2 gating from direct cytosolic Ca²+ regulation.


The provision of dental implants in the National Health Service Hospital dental services--a national questionnaire.
To assess the activity of consultants in restorative dentistry in the United Kingdom in the provision of osseointegrated dental implants within the National Health Service Hospital service and to evaluate their attitudes concerning the relevant medical and oral factors considered in patient selection for implant treatment. Anonymous postal questionnaire in the United Kingdom. Consultants in restorative dentistry. Out of the sample of 145, 109 consultants (75%) completed the questionnaire in 1999. 54 of the 109 consultants (49.5%) are involved in the provision of osseointegrated implant treatment, treating an average of 29 cases/year (range 2-150). However, over one third of the respondents treated 10 or less cases/year. 89% worked with oral surgeons as an implant team. 68% used Branemark (Nobel Biocare) implants as their main system. The majority of consultants felt that smoking, psychoses and previous irradiation were the most important medical factors that contra-indicated implant retained restorations whilst untreated periodontitis, poor oral hygiene and uncontrolled caries were the most important oral contra-indications. Many centres were experiencing significant problems with the funding of implant treatment with one centre receiving no funding. The implications for patient care and specialist training are discussed. There is a marked variation in the number of patients treated with endosseous dental implants within the United Kingdom National Health Service hospitals. Many consultants treat 10 or fewer patients each year. In the main, there is agreement about the factors that contra-indicate implant treatment; these are in line with national guidelines.


The transfer of adaptive immunity to CMV during hematopoietic stem cell transplantation is dependent on the specificity and phenotype of CMV-specific T cells in the donor.
The successful reconstitution of adaptive immunity to human cytomegalovirus (CMV) in hematopoietic stem cell transplantation (HSCT) recipients is central to the reduction of viral reactivation-related morbidity and mortality. Here, we characterized the magnitude, specificity, phenotype, function, and clonotypic composition of CMV-specific T-cell responses in 18 donor-recipient pairs both before and after HSCT. The principal findings were: (1) the specificity of CMV-specific T-cell responses in the recipient after HSCT mirrors that in the donor; (2) the maintenance of these targeting patterns reflects the transfer of epitope-specific T-cell clonotypes from donor to recipient; (3) less differentiated CD27(+)CD57(-) CMV-specific memory T cells are more likely to persist in the recipient after HSCT compared with more terminally differentiated CD27(-) CD57(+) CMV-specific memory T cells; (4) the presence of greater numbers of less differentiated CD8(+) CMV-specific T cells in the donor appears to confer protection against viral reactivation in the recipient after HSCT; and (5) CMV-specific T cells acquire a more differentiated phenotype and a restricted functional profile after HSCT. Overall, these findings define the immunologic factors that influence the successful adoptive transfer of antigen-specific T-cell immunity during HSCT, which enables the identification of recipients at particular risk of CMV reactivation after HSCT.


Group II and III mGluRs-mediated presynaptic inhibition of EPSCs recorded from hippocampal interneurons of CA1 stratum lacunosum moleculare.
We have studied the effects of groups II and III metabotropic glutamate receptor (mGluR) activation on excitatory responses recorded from hippocampal interneurons of CA1 stratum lacunosum moleculare (SLM). Excitatory postsynaptic currents (EPSCs) evoked by stimulation of the perforant pathway were reduced either by the group II mGluR agonist LY354740 (50-100 nM, 49.1+/-5.7% of control) or by the group III mGluR agonist l-2-amino-4-phosphonobutyric acid (l-AP4) (50 microM, 36.8+/-4.4% of control). Both drugs significantly enhanced paired-pulse facilitation of the EPSCs. Furthermore, both 100 nM LY354740 and 50 microM l-AP4 reduced the frequency, but not the amplitude, of miniature excitatory synaptic currents (mEPSCs), recorded in the presence of 1 microM TTX and 50 microM picrotoxin, or EPSCs evoked by perforant pathway stimulation in the presence of 2.5 mM Sr2+. The broad-spectrum mGluR antagonist LY341495 (10-50 microM) did not affect test EPSCs elicited 210 ms after stimulation at 100 Hz. At network level, 1-5 microM LY354740 significantly reduced the power of gamma frequency oscillations induced by 20 microM carbachol, 600 nM kainate and 5 mM K+ in hippocampal CA1 area. Our results show powerful modulation of excitatory transmission impinging on interneurons of CA1 SLM by presynaptic group II or III mGluRs.


Integer Ratio Priors on Musical Rhythm Revealed Cross-culturally by Iterated Reproduction.
Probability distributions over external states (priors) are essential to the interpretation of sensory signals. Priors for cultural artifacts such as music and language remain largely uncharacterized, but likely constrain cultural transmission, because only those signals with high probability under the prior can be reliably reproduced and communicated. We developed a method to estimate priors for simple rhythms via iterated reproduction of random temporal sequences. Listeners were asked to reproduce random "seed" rhythms; their reproductions were fed back as the stimulus and over time became dominated by internal biases, such that the prior could be estimated by applying the procedure multiple times. We validated that the measured prior was consistent across the modality of reproduction and that it correctly predicted perceptual discrimination. We then measured listeners' priors over the entire space of two- and three-interval rhythms. Priors in US participants showed peaks at rhythms with simple integer ratios and were similar for musicians and non-musicians. An analogous procedure produced qualitatively different results for spoken phrases, indicating some specificity to music. Priors measured in members of a native Amazonian society were distinct from those in US participants but also featured integer ratio peaks. The results do not preclude biological constraints favoring integer ratios, but they suggest that priors on musical rhythm are substantially modulated by experience and may simply reflect the empirical distribution of rhythm that listeners encounter. The proposed method can efficiently map out a high-resolution view of biases that shape transmission and stability of simple reproducible patterns within a culture.


Double trouble at high density: cross-level test of resource-related adaptive plasticity and crowding-related fitness.
Population size is often regulated by negative feedback between population density and individual fitness. At high population densities, animals run into double trouble: they might concurrently suffer from overexploitation of resources and also from negative interference among individuals regardless of resource availability, referred to as crowding. Animals are able to adapt to resource shortages by exhibiting a repertoire of life history and physiological plasticities. In addition to resource-related plasticity, crowding might lead to reduced fitness, with consequences for individual life history. We explored how different mechanisms behind resource-related plasticity and crowding-related fitness act independently or together, using the water flea Daphnia magna as a case study. For testing hypotheses related to mechanisms of plasticity and crowding stress across different biological levels, we used an individual-based population model that is based on dynamic energy budget theory. Each of the hypotheses, represented by a sub-model, is based on specific assumptions on how the uptake and allocation of energy are altered under conditions of resource shortage or crowding. For cross-level testing of different hypotheses, we explored how well the sub-models fit individual level data and also how well they predict population dynamics under different conditions of resource availability. Only operating resource-related and crowding-related hypotheses together enabled accurate model predictions of D. magna population dynamics and size structure. Whereas this study showed that various mechanisms might play a role in the negative feedback between population density and individual life history, it also indicated that different density levels might instigate the onset of the different mechanisms. This study provides an example of how the integration of dynamic energy budget theory and individual-based modelling can facilitate the exploration of mechanisms behind the regulation of population size. Such understanding is important for assessment, management and the conservation of populations and thereby biodiversity in ecosystems.


Damping effects on the response of maxillary incisor subjected to a traumatic impact force: a nonlinear finite element analysis.
The aim of this study was to evaluate the effects of damping on stress concentration in an impacted incisor. Damping ratios of maxillary incisors were tested using an in vivo modal testing method. A finite element model of the upper central incisor was established for dental trauma analysis. To assess the effect of damping properties on induced stresses in the traumatized incisors, equivalent stresses in the finite element model with various damping ratios were calculated for comparison. The mechanisms of cushioning properties of the upper incisors on traumatic injuries were assessed by profiling the stress distributions in the incisor model sequentially with time. The measured damping ratio of maxillary incisors was 0.146+/-0.037. When the incisor was subjected to an impact force, high stresses were concentrated at the labial and lingual incisor edges, cervical ridge, and the area around root apex. When the damping ratios of the incisor model were set at 10- and 50-fold of the measured values, the peak stresses induced near the impact site of the incisor model were reduced from 24.0 to 23.2 and 15.9 MPa, respectively. On the other hand, the peak stress lagged and the stress existence period increased when the damping properties were taken into consideration. Damping properties of teeth provide protection to the tooth during traumatic injury by decreasing the peak stress magnitude due to release of strain energy over a longer period.


Venous thromboembolism following primary total knee arthroplasty.
Total knee arthroplasty is associated with a high postoperative incidence of venous thromboembolism (VTE). Without thromboprophylaxis, as many as 80% of patients may develop deep vein thrombosis (DVT). Evidence suggests that pharmacological prophylaxis may not be offered due to concern of bleeding with anticoagulants. We retrospectively reviewed the clinical incidence of VTE and thromboprophylactic practice patterns over a 9-year period. Patient baseline characteristics, diagnosis, VTE risk factors, prophylactic modalities (mechanical and pharmacological), operation duration, type of prosthesis and fixation, mode of anesthesia, hospital length of stay (LOS) and postoperative complications with particular attention to suspected DVT and/or pulmonary embolism (PE) were analysed. Male to female ratio was 1:2.3, median age 71 (interquartile range, IQR: 65-77) years and hospital LOS of 8 (IQR: 7-11) days. The in-hospital VTE incidence was 3.9% (95% confidence interval, CI: 2.2-6.8%) with a possibly underestimated 3-month rate of 5.7% (95% CI: 1.6-18.6%). In-hospital proximal DVT incidence was 0.7% (95% CI: 0.2-2.5%) and 2.9% (95% CI: 0.5-14.5%) at 3 months. Non fatal PE was 0.7% (95% CI: 1.2-5%). DVT rate was higher with cemented prostheses (P=0.008), with a greater rate of bleeding when heparin was commenced preoperatively (P=0.001). The rate of in-hospital VTE was kept relatively low with the use of prophylactic protocols with all patients receiving prophylaxis. Given our one and a half and four-fold increase in the out of hospital VTE and proximal DVT incidence, consideration should be given to continued prophylaxis beyond hospitalization in this high-risk group of patients.


Hypercoagulation and thrombophilia in liver disease.
A complex balance exists between endogenous procoagulants and the anticoagulant system in liver disease patients. Hypercoagulable events occur in cirrhosis patients despite the well-known bleeding diathesis of liver disease. These events may be clinically evident, such as in portal vein thrombosis or pulmonary embolism, but these conditions may also be a silent contributor to certain disease states, such as portopulmonary hypertension or parenchymal extinction with liver atrophy as well as thrombosis of extracorporeal circuits in dialysis or liver assist devices. Moreover, liver disease-related hypercoagulability may contribute to vascular disease in the increasingly common condition of non-alcoholic fatty liver disease. Despite the incidence of these problems, there are few widely accessible and practical laboratory tests to evaluate the risk of a hypercoagulable event in cirrhosis patients. Furthermore, there is little research on the use of commonly accepted anticoagulants in patients with liver disease. This article is a result of an international symposium on coagulation disorders in liver disease and addresses several areas of specific interest in hypercoagulation in liver disease. Critical areas lacking clinical information are highlighted and future areas of research interest are defined with an aim to foster clinical research in this field.


Three-dimensional extracellular matrix engineering in the nervous system.
Growing neurites are guided through their environment during development and regeneration via different cellular and extracellular matrix (ECM) molecular cues. To mimic cell-matrix interactions, a three-dimensional (3D) hydrogel-based ECM equivalent containing a covalently immobilized laminin oligopeptide sequence was designed to facilitate nerve regeneration. This study illustrates that the oligopeptide domain CDPGYIGSR covalently linked to an agarose gel as a bioartificial 3D substrate successfully supports neurite outgrowth from dorsal root ganglia (DRG) in vitro. The specificity of the neurite promoting activity was illustrated through the inhibition of neurite outgrowth from DRG in a CDPGYIGSR-derivatized gel in the presence of solubilized CDPGYIGSR peptide. Gels derivatized with CDPGYIGSK and CDPGRGSYI peptides stimulated a smaller increase of neurite outgrowth. In vivo experiments revealed the capability of a CDPGYIGSR-derivatized gel to enhance nerve regeneration in a transected rat dorsal root model compared to an underivatized gel, a CDPGRGSYI gel, and saline-filled nerve guidance channels. These data suggest the feasibility of a 3D hydrogel-based ECM equivalent capable of enhancing neurite outgrowth in vitro and in vivo.


Cloning and characterization of cyclooxygenase-1b (putative cyclooxygenase-3) in rat.
A splice variant of cyclooxygenase-1 (COX-1), COX-1b (previously termed as COX-3), has been identified in canine tissues as an acetaminophen-sensitive isoform, but the sequence of COX-1b mRNA and the encoded protein are not known in rats. We cloned and sequenced rat COX-1b mRNA from cerebral endothelial cells. Sequence analysis indicated that the 98-base pair intron-1 of COX-1 gene remains unprocessed in the COX-1b mRNA, causing a frameshift mutation and a 127-amino acid open reading frame with no sequence similarity with known cyclooxygenases. Transient and permanent transfection of COS-7 cells with a vector containing the rat COX-1b cDNA resulted in synthesis of a protein of the expected size. We generated an affinity-purified polyclonal antibody against the rat COX-1b protein. Western blot analysis of rat tissues using this antibody demonstrated the likely existence of rat COX-1b protein in vivo with the highest expression in heart, kidney, and neuronal tissues. Our results on both stable and on transiently transfected COS-7 cells suggest that rat COX-1b does not have cyclooxygenase activity and does not have any effect on the inhibition of prostaglandin production by acetaminophen. Because this protein has a completely different amino acid sequence than COX-1 and COX-2 and it does not have cyclooxygenase activity, we suggest a name cyclooxygenase variant protein to distinguish it from the known prostaglandin-synthesizing cyclooxygenase isoforms.


Histologic changes after vertebroplasty.
Vertebroplasty with use of polymethylmethacrylate cement is gaining popularity in the treatment of some specific painful lesions of the spine. It remains unclear, however, what possible side effects this type of cement might have upon the vertebral body. We performed a histologic and radiographic analysis of the end plate and disc to determine whether there was a difference between vertebroplasty with polymethylmethacrylate cement and vertebroplasty with calcium phosphate cement in the surrounding tissue of the goat spine. Furthermore, we assessed whether a defect in the end plate, simulating end-plate fracture and allowing for direct contact of cement with disc tissue, had any effect on end-plate or disc degeneration. Twenty-four mature goats were divided between two follow-up periods (six weeks and six months). All animals underwent a bilateral transpedicular vertebroplasty at two lumbar levels, where one of the following treatments was applied: vertebroplasty with calcium phosphate cement with or without an end-plate defect, and vertebroplasty with polymethylmethacrylate cement with or without an end-plate defect. The effect of the various treatments on the integrity of the intervertebral disc, end plate, and surrounding tissue was examined with semiquantitative histologic analysis and radiography. No sign of disc or end-plate degeneration was seen in any of the analyzed sections. The mean disc height did not decrease from the postoperative period to the time that the animals were killed in any group, thereby supporting the histologic findings. A mild inflammatory reaction was found in four vertebral bodies in the polymethylmethacrylate groups only. Calcium phosphate cement and polymethylmethacrylate cement both seem to be adequate bone-void fillers in terms of biological behavior in the vertebral body.


Constitutive shedding of both p55 and p75 murine TNF receptors in vivo.
Recently, we reported the generation and characterization of hamster mAbs specific for the murine p55 and p75 TNF receptors. Upon characterizing these mAbs in vivo, we discovered that administration of TNF receptor-specific mAb to normal mice resulted in the linear accumulation of the appropriate class of soluble TNF receptor in the circulation. The mechanism underlying soluble receptor accumulation was found to be due to an abrogation of the clearance of constitutively shed soluble receptor by receptor-specific mAb. Levels of p55 or p75 accumulated in the presence of nonblocking, nonagonistic TNF receptor mAb were capable of inhibiting murine TNF-induced responses in vivo. These results document that both p55 and p75 are constitutively shed in substantial amounts in vivo and suggest that the process of constitutive TNF receptor shedding plays an important role in regulating TNF activity under physiologic conditions.


Emended description of Actinomyces naeslundii and descriptions of Actinomyces oris sp. nov. and Actinomyces johnsonii sp. nov., previously identified as Actinomyces naeslundii genospecies 1, 2 and WVA 963.
Actinomyces naeslundii is an important early colonizer in the oral biofilm and consists of three genospecies (1, 2 and WVA 963) which cannot be readily differentiated using conventional phenotypic testing or on the basis of 16S rRNA gene sequencing. We have investigated a representative collection of type and reference strains and clinical and oral isolates (n=115) and determined the partial gene sequences of six housekeeping genes (atpA, rpoB, pgi, metG, gltA and gyrA). These sequences identified the three genospecies and differentiated them from Actinomyces viscosus isolated from rodents. The partial sequences of atpA and metG gave best separation of the three genospecies. A. naeslundii genospecies 1 and 2 formed two distinct clusters, well separated from both genospecies WVA 963 and A. viscosus. Analysis of the same genes in other oral Actinomyces species (Actinomyces gerencseriae, A. israelii, A. meyeri, A. odontolyticus and A. georgiae) indicated that, when sequence data were obtained, these species each exhibited <90 % similarity with the A. naeslundii genospecies. Based on these data, we propose the name Actinomyces oris sp. nov. (type strain ATCC 27044(T) =CCUG 34288(T)) for A. naeslundii genospecies 2 and Actinomyces johnsonii sp. nov. (type strain ATCC 49338(T) =CCUG 34287(T)) for A. naeslundii genospecies WVA 963. A. naeslundii genospecies 1 should remain as A. naeslundii sensu stricto, with the type strain ATCC 12104(T) =NCTC 10301(T) =CCUG 2238(T).


Characterization of dengue virus complex-specific neutralizing epitopes on envelope protein domain III of dengue 2 virus.
The surface of the mature dengue virus (DENV) particle is covered with 180 envelope (E) proteins arranged as homodimers that lie relatively flat on the virion surface. Each monomer consists of three domains (ED1, ED2, and ED3), of which ED3 contains the critical neutralization determinant(s). In this study, a large panel of DENV-2 recombinant ED3 mutant proteins was used to physically and biologically map the epitopes of five DENV complex-specific monoclonal antibodies (MAbs). All five MAbs recognized a single antigenic site that includes residues K310, I312, P332, L389, and W391. The DENV complex antigenic site was located on an upper lateral surface of ED3 that was distinct but overlapped with a previously described DENV-2 type-specific antigenic site on ED3. The DENV complex-specific MAbs required significantly higher occupancy levels of available ED3 binding sites on the virion, compared to DENV-2 type-specific MAbs, in order to neutralize virus infectivity. Additionally, there was a great deal of variability in the neutralization efficacy of the DENV complex-specific MAbs with representative strains of the four DENVs. Overall, the differences in physical binding and potency of neutralization observed between DENV complex- and type-specific MAbs in this study demonstrate the critical role of the DENV type-specific antibodies in the neutralization of virus infectivity.


Associations between hippocampal morphometry and neuropathologic markers of Alzheimer's disease using 7 T MRI.
Hippocampal atrophy, amyloid plaques, and neurofibrillary tangles are established pathologic markers of Alzheimer's disease. We analyzed the temporal lobes of 9 Alzheimer's dementia (AD) and 7 cognitively normal (NC) subjects. Brains were scanned post-mortem at 7 Tesla. We extracted hippocampal volumes and radial distances using automated segmentation techniques. Hippocampal slices were stained for amyloid beta (Aβ), tau, and cresyl violet to evaluate neuronal counts. The hippocampal subfields, CA1, CA2, CA3, CA4, and subiculum were manually traced so that the neuronal counts, Aβ, and tau burden could be obtained for each region. We used linear regression to detect associations between hippocampal atrophy in 3D, clinical diagnosis and total as well as subfield pathology burden measures. As expected, we found significant correlations between hippocampal radial distance and mean neuronal count, as well as diagnosis. There were subfield specific associations between hippocampal radial distance and tau in CA2, and cresyl violet neuronal counts in CA1 and subiculum. These results provide further validation for the European Alzheimer's Disease Consortium Alzheimer's Disease Neuroimaging Initiative Center Harmonized Hippocampal Segmentation Protocol (HarP).


Biomass viability: An experimental study and the development of an empirical mathematical model for submerged membrane bioreactor.
This study investigates the influence of key biomass parameters on specific oxygen uptake rate (SOUR) in a sponge submerged membrane bioreactor (SSMBR) to develop mathematical models of biomass viability. Extra-cellular polymeric substances (EPS) were considered as a lumped parameter of bound EPS (bEPS) and soluble microbial products (SMP). Statistical analyses of experimental results indicate that the bEPS, SMP, mixed liquor suspended solids and volatile suspended solids (MLSS and MLVSS) have functional relationships with SOUR and their relative influence on SOUR was in the order of EPS>bEPS>SMP>MLVSS/MLSS. Based on correlations among biomass parameters and SOUR, two independent empirical models of biomass viability were developed. The models were validated using results of the SSMBR. However, further validation of the models for different operating conditions is suggested.


Some properties of the optical resonances in a single subwavelength slit.
Diffraction of TM-polarized waves by a slit in a thick screen of infinite conductivity is treated. The case of an arbitrary incident beam wave is considered. We study the resonances that appear when the wavelength of the incident beam wave is larger than the slit width, i.e., the subwavelength regime where a one-mode model for the slit can be considered. High anomalous values (resonances) of the transmission coefficient, the angular diffracted energy, and the magnetic field within the slit are analyzed. A simple linear relationship to determine the resonant wavelengths is proposed. We show that the transmission coefficient, the normal diffracted energy, and the magnetic field within the cavity are linear functions of the resonant wavelength and the thickness of the screen. Additionally and surprisingly, we reveal that under certain conditions the incident beam wave via the diffraction can give a suppressed light transmission; i.e., a minimum in the transmission is obtained where a maximum is expected.


Enterohaemorrhagic Escherichia coli Tir requires a C-terminal 12-residue peptide to initiate EspF-mediated actin assembly and harbours N-terminal sequences that influence pedestal length.
Enterohaemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) both utilize type III secretion systems that translocate the effector protein Tir into the plasma membrane of mammalian cells in order to stimulate localized actin assembly into 'pedestals'. The Tir molecule that EPEC delivers is phosphorylated within its C-terminus on tyrosine-474, and a clustered 12-residue phosphopeptide encompassing this residue initiates an efficient signalling cascade that triggers actin polymerization. In addition to Y474, tyrosine-454 of EPEC Tir is phosphorylated, although inefficiently, and promotes actin polymerization at low levels. In contrast to EPEC Tir, EHEC Tir lacks Y474 and triggers pedestal formation in a phosphotyrosine-independent manner by interacting with an additional effector protein, EspF(U). To identify EHEC Tir sequences that regulate localized actin assembly, we circumvented the strict requirements for type III translocation and directly expressed Tir derivatives in mammalian cells by transfection. Infection of Tir-expressing cells with a Tir-deficient EHEC strain demonstrated that ectopically expressed Tir localizes to the plasma membrane, is modified by mammalian serine-threonine kinases and is fully functional for actin pedestal formation. Removal of portions of the cytoplasmic N-terminus of Tir resulted in the generation of abnormally long pedestals, indicating that this region of EHEC Tir influences pedestal length. In the presence of the entire N-terminal domain, a 12-residue peptide from the C-terminus of EHEC Tir is both necessary and sufficient to recruit EspF(U) and initiate actin pedestal formation. This peptide encompasses the portion of EHEC Tir analogous to the EPEC Tir-Y454 region and is present within the Tir molecules of all pedestal-forming bacteria, suggesting that this sequence harbours a conserved signalling function.


When knowing becomes remembering: individual differences in susceptibility to suggestion.
In two experiments, the authors explored factors that might influence a person's tendency to make source-monitoring errors about autobiographical memories. In the first experiment, undergraduates retrieved a memory from childhood (a) that was known about but not remembered, (b) that was remembered, or (c) for which they were unsure of their memory's source. After writing down the memory, experimental groups listened to a guided visualization tape and answered questions about the event--interventions designed to help them focus on details of their memory. Controls also retrieved and wrote down a memory; however, instead of visualizing the memory, they were instructed to conduct a visual search task. Results indicated that guided visualization led participants to rate known memories closer to remembered events. A second experiment examined individual difference variables that might be related to this know-to-remember shift. Results indicated that extraversion, external locus of control, a memory that conveyed fear, and overall affective content predicted this rating. The applicability of these findings to the psychotherapy process is discussed.


CD44(+) cells have cancer stem cell-like properties in nasopharyngeal carcinoma.
A subpopulation of cells within a tumor appears to have the exclusive ability to initiate tumors, self-renew, and differentiate. These "cancer stem cells" (CSCs) are CD44(+) in several epithelial malignancies. We examined the potential of CD44 to identify the CSC population in nasopharyngeal carcinoma (NPC). C666, an Epstein-Barr virus-positive (EBV(+) ) human NPC cell line, was stained for CD44 and sorted by fluorescence-activated cell sorting (FACS). CD44(+) and CD44(-) subpopulations were evaluated for (1) proliferative potential, (2) ability to differentiate, (3) expression of markers of epithelial-to-mesenchymal transition (EMT) and EBV genes, and (4) the ability to initiate tumors in vivo. Immunocompromised mice were injected with CD44(+) and CD44(-) populations to assess the tumor-initiating capacity. Immunohistochemistry for CD44 was performed on an 87-patient tissue microarray (TMA), and clinical correlations were examined. Heterogeneous expression of CD44 was seen among C666 cells. CD44(+) cells differentiated into CD44(-) cells, indicating a hierarchical relationship. Further, CD44(+) cells exhibited a more robust tumor-initiating capacity in the xenograft model. However, no differences were seen in proliferation rates in vitro, EBV gene expression, or expression of EMT markers between CD44(+) and CD44(-) subsets. Patient tumors were heterogeneous for CD44 staining, and a trend toward an association between CD44 expression and clinical outcome was observed. NPC contains a CD44(+) subpopulation with features consistent with CSCs. There was a trend toward an association between CD44 expression within NPC tumors and decreased time to local failure/relapse in patients.


The Role of Visual-Spatial Abilities in Dyslexia: Age Differences in Children's Reading?
Reading is a highly complex process in which integrative neurocognitive functions are required. Visual-spatial abilities play a pivotal role because of the multi-faceted visual sensory processing involved in reading. Several studies show that children with developmental dyslexia (DD) fail to develop effective visual strategies and that some reading difficulties are linked to visual-spatial deficits. However, the relationship between visual-spatial skills and reading abilities is still a controversial issue. Crucially, the role that age plays has not been investigated in depth in this population, and it is still not clear if visual-spatial abilities differ across educational stages in DD. The aim of the present study was to investigate visual-spatial abilities in children with DD and in age-matched normal readers (NR) according to different educational stages: in children attending primary school and in children and adolescents attending secondary school. Moreover, in order to verify whether visual-spatial measures could predict reading performance, a regression analysis has been performed in younger and older children. The results showed that younger children with DD performed significantly worse than NR in a mental rotation task, a more-local visual-spatial task, a more-global visual-perceptual task and a visual-motor integration task. However, older children with DD showed deficits in the more-global visual-perceptual task, in a mental rotation task and in a visual attention task. In younger children, the regression analysis documented that reading abilities are predicted by the visual-motor integration task, while in older children only the more-global visual-perceptual task predicted reading performances. Present findings showed that visual-spatial deficits in children with DD were age-dependent and that visual-spatial abilities engaged in reading varied across different educational stages. In order to better understand their potential role in affecting reading, a comprehensive description and a multi-componential evaluation of visual-spatial abilities is needed with children with DD.


Current State of Electronic Consent Processes in Behavioral Health: Outcomes from an Observational Study.
An integral element of value-based care is care team access to both physical and behavioral health data. Data release processes in both environments are governed by federal and state statutes. The requirements for obtaining consent are complex and often confusing. Little is known about the consent processes and practices in the behavioral health setting, specifically how patients and surrogates engage in the process and their interactions with electronic consent tools. This study analyzes the consent processes from the patient perspective at two community behavioral health clinics. Outcomes include description of the processes using electronic consent, workflows and consenter-provider interactions. Conclusions include need to streamline and standardize consent technologies and improve consenter engagement. This study supports the development of an electronic consent tool, My Data Choices (MDC), funded by the National Institute of Mental Health, that offers individuals with behavioral health conditions more control over their medical records.


Substrates of IAP ubiquitin ligases identified with a designed orthogonal E3 ligase, the NEDDylator.
Inhibitors of Apoptosis Protein (IAPs) are guardian ubiquitin ligases that keep classic proapoptotic proteins in check. Systematic identification of additional IAP substrates is challenged by the heterogeneity and sheer number of ubiquitinated proteins (>5,000). Here we report a powerful catalytic tagging tool, the NEDDylator, which fuses a NEDD8 E2-conjugating enzyme, Ubc12, to the ubiquitin ligase, XIAP or cIAP1. This permits transfer of the rare ubiquitin homolog NEDD8 to the ubiquitin E3 substrates, allowing them to be efficiently purified for LC-MS/MS identification. We have identified >50 potential IAP substrates of both cytosolic and mitochondrial origin that bear hallmark N-terminal IAP binding motifs. These substrates include the recently discovered protein phosphatase PGAM5, which we show is proteolytically processed, accumulates in cytosol during apoptosis, and sensitizes cells to death. These studies reveal mechanisms and antagonistic partners for specific IAPs, and provide a powerful technology for labeling binding partners in transient protein-protein complexes.


Meningiomas invading the superior sagittal sinus: surgical experience in 108 cases.
Radical resection of meningiomas invading the superior sagittal sinus (SSS) presents several hazards. Some surgeons consider SSS invasion a contraindication for complete resection, and others advocate total resection with venous reconstruction. There is a lack of published large series to provide definitive guidelines for the surgical treatment of these complex cases. We report our 15-year experience with surgery of parasagittal meningiomas invading the SSS. Between 1986 and 2001, 108 patients (73 women, 35 men; age range, 22-83 yr; mean age, 56.2 yr) underwent surgery at the Neurological Institute "C. Besta" of Milan for tumors invading the SSS. Parasagittal meningiomas not invading the SSS were excluded from this series. Simpson Grade I to II removal was achieved in 100 patients. Thirty patients with meningiomas totally occluding the SSS had complete resection of the encased portion of the sinus. Histological examination revealed 86 benign (79.6%), 16 atypical (14.8%), and 4 malignant (3.7%) meningiomas along with 2 hemangiopericytomas. There were two perioperative deaths. Serious complications included brain swelling (nine patients; 8.3%) and postoperative hematoma (two patients; 1.85%). Follow-up ranged from 19 to 223 months (mean, 79.5 mo). One patient was lost to follow-up. Tumors recurred in 15 patients (13.9%). After multivariate analysis, histological type, tumor size, and Simpson grade were confirmed as significant independent prognostic factors for recurrence. On the basis of our results, we conclude that if the sinus is partially invaded, it can be opened to obtain as complete a resection as possible and to attempt to preserve the patency of the sinus. If the sinus is obstructed, the portion of the sinus involved can be resected completely. In both situations, extreme care is vital to preservation of cortical veins, which may offer important collateral drainage. With our approach, good results are achieved and it is not necessary to reconstruct the sinus.


Immune evasion mechanisms in colorectal cancer liver metastasis patients vaccinated with TroVax (MVA-5T4).
We have recently reported the results of a phase II trial in which two TroVax [modified vaccinia ankara (MVA) encoding the tumour antigen 5T4] vaccinations were given to patients both pre- and post-surgical resection of liver metastases secondary to colorectal cancer (CRC). 5T4-specific cellular responses were assessed at the entry and 2 weeks after each vaccination by proliferation of fresh lymphocytes and ELISA for antibody responses; 18 from the 19 CRC patients mounted a 5T4-specific cellular and/or humoral response. Here, we present a comparison of individual and between patient responses over the course of the treatments using cryopreserved peripheral blood mononuclear cells (PBMC) samples from the baseline until after the fourth vaccination at 14 weeks. Assays used were proliferation assay with 5T4-Fc fusion protein, overlapping 32mer 5T4 peptides, MVA-LacZ and MVA-5T4 infected autologous monocytes. Responses to 5T4 protein or one or more peptide pools were pre-existing in 12/20 patients and subsequently 10 and 12 patients showed boosted and/or de novo responses, respectively. Cumulatively, 13/20 patients showed proliferative responses by week 14. We also assessed the levels of systemic T regulatory cells, plasma cytokine levels, phenotype of tumour-infiltrating lymphocytes including T regulatory cells and tumour HLA class I loss of expression. More than half of the patients showed phenotypes consistent with relative immune suppression and/or escape highlighting the complexity of positive and negative factors challenging any simple correlation with clinical outcome.


A transcriptional fingerprint of estrogen in human breast cancer predicts patient survival.
Estrogen signaling plays an essential role in breast cancer progression, and estrogen receptor (ER) status has long been a marker of hormone responsiveness. However, ER status alone has been an incomplete predictor of endocrine therapy, as some ER+ tumors, nevertheless, have poor prognosis. Here we sought to use expression profiling of ER+ breast cancer cells to screen for a robust estrogen-regulated gene signature that may serve as a better indicator of cancer outcome. We identified 532 estrogen-induced genes and further developed a 73-gene signature that best separated a training set of 286 primary breast carcinomas into prognostic subtypes by stepwise cross-validation. Notably, this signature predicts clinical outcome in over 10 patient cohorts as well as their respective ER+ subcohorts. Further, this signature separates patients who have received endocrine therapy into two prognostic subgroups, suggesting its specificity as a measure of estrogen signaling, and thus hormone sensitivity. The 73-gene signature also provides additional predictive value for patient survival, independent of other clinical parameters, and outperforms other previously reported molecular outcome signatures. Taken together, these data demonstrate the power of using cell culture systems to screen for robust gene signatures of clinical relevance.


Polysaccharides from Ganoderma lucidum attenuate microglia-mediated neuroinflammation and modulate microglial phagocytosis and behavioural response.
Ganoderma lucidum (GL) has been widely used in Asian countries for hundreds of years to promote health and longevity. The pharmacological functions of which had been classified, including the activation of innate immune responses, suppression of tumour and modulation of cell proliferations. Effective fractions of Ganoderma lucidum polysaccharides (GLP) had already been reported to regulate the immune system. Nevertheless, the role of GLP in the microglia-mediated neuroinflammation has not been sufficiently elucidated. Further, GLP effect on microglial behavioural modulations in correlation with the inflammatory responses remains to be unravelled. The aim of this work was to quantitatively analyse the contributions of GLP on microglia. The BV2 microglia and primary mouse microglia were stimulated by lipopolysaccharides (LPS) and amyloid beta42 (Aβ42) oligomer, respectively. Investigation on the effect of GLP was carried by quantitative determination of the microglial pro- and anti-inflammatory cytokine expressions and behavioural modulations including migration, morphology and phagocytosis. Analysis of microglial morphology and phagocytosis modulations was confirmed in the zebrafish brain. Quantitative results revealed that GLP down-regulates LPS- or Aβ-induced pro-inflammatory cytokines and promotes anti-inflammatory cytokine expressions in BV-2 and primary microglia. In addition, GLP attenuates inflammation-related microglial migration, morphological alterations and phagocytosis probabilities. We also showed that modulations of microglial behavioural responses were associated with MCP-1 and C1q expressions. Overall, our study provides an insight into the GLP regulation of LPS- and Aβ-induced neuroinflammation and serves an implication that the neuroprotective function of GLP might be achieved through modulation of microglial inflammatory and behavioural responses.


Penumbral imaging and functional outcome in patients with anterior circulation ischaemic stroke treated with endovascular thrombectomy versus medical therapy: a meta-analysis of individual patient-level data.
CT perfusion (CTP) and diffusion or perfusion MRI might assist patient selection for endovascular thrombectomy. We aimed to establish whether imaging assessments of irreversibly injured ischaemic core and potentially salvageable penumbra volumes were associated with functional outcome and whether they interacted with the treatment effect of endovascular thrombectomy on functional outcome. In this systematic review and meta-analysis, the HERMES collaboration pooled patient-level data from all randomised controlled trials that compared endovascular thrombectomy (predominantly using stent retrievers) with standard medical therapy in patients with anterior circulation ischaemic stroke, published in PubMed from Jan 1, 2010, to May 31, 2017. The primary endpoint was functional outcome, assessed by the modified Rankin Scale (mRS) at 90 days after stroke. Ischaemic core was estimated, before treatment with either endovascular thrombectomy or standard medical therapy, by CTP as relative cerebral blood flow less than 30% of normal brain blood flow or by MRI as an apparent diffusion coefficient less than 620 μm2/s. Critically hypoperfused tissue was estimated as the volume of tissue with a CTP time to maximum longer than 6 s. Mismatch volume (ie, the estimated penumbral volume) was calculated as critically hypoperfused tissue volume minus ischaemic core volume. The association of ischaemic core and penumbral volumes with 90-day mRS score was analysed with multivariable logistic regression (functional independence, defined as mRS score 0-2) and ordinal logistic regression (functional improvement by at least one mRS category) in all patients and in a subset of those with more than 50% endovascular reperfusion, adjusted for baseline prognostic variables. The meta-analysis was prospectively designed by the HERMES executive committee, but not registered. We identified seven studies with 1764 patients, all of which were included in the meta-analysis. CTP was available and assessable for 591 (34%) patients and diffusion MRI for 309 (18%) patients. Functional independence was worse in patients who had CTP versus those who had diffusion MRI, after adjustment for ischaemic core volume (odds ratio [OR] 0·47 [95% CI 0·30-0·72], p=0·0007), so the imaging modalities were not pooled. Increasing ischaemic core volume was associated with reduced likelihood of functional independence (CTP OR 0·77 [0·69-0·86] per 10 mL, pinteraction=0·29; diffusion MRI OR 0·87 [0·81-0·94] per 10 mL, pinteraction=0·94). Mismatch volume, examined only in the CTP group because of the small numbers of patients who had perfusion MRI, was not associated with either functional independence or functional improvement. In patients with CTP with more than 50% endovascular reperfusion (n=186), age, ischaemic core volume, and imaging-to-reperfusion time were independently associated with functional improvement. Risk of bias between studies was generally low. Estimated ischaemic core volume was independently associated with functional independence and functional improvement but did not modify the treatment benefit of endovascular thrombectomy over standard medical therapy for improved functional outcome. Combining ischaemic core volume with age and expected imaging-to-reperfusion time will improve assessment of prognosis and might inform endovascular thrombectomy treatment decisions. Medtronic.


Control of glial cell volume in anoxia. In vitro studies on ischemic cell swelling.
Volume regulation of C6 glial cells was studied in anoxia in vitro to improve the understanding of ischemic cell swelling in the brain. Contrary to in vivo conditions, anoxia or anoxia plus iodoacetate for additional inhibition of anaerobic energy metabolism did not induce glial swelling. However, intracellular K+ was markedly decreased while intracellular Na+ increased. Induction of energy failure by anoxia plus iodoacetate was found to prevent the regulatory volume decrease on hyposmotic exposure of the cells, which is regularly observed in normoxic control conditions. Hyposmotic exposure in anoxia plus iodoacetate led only to an initial tendency of cell volume normalization followed by secondary cell swelling. This was associated with a net increase of intracellular Na+ that may explain the failure of volume regulation under these circumstances. Maintenance of a normal glial cell size during complete energy deprivation by anoxia plus iodoacetate in isotonic medium strongly indicates that energy failure per se does not suffice to induce cell swelling. Cell swelling in cerebral ischemia in vivo thus is likely to require additional mechanisms, most likely an increase of membrane permeability to Na+, which may be caused by release and accumulation of excitotoxins such as glutamate or by an extracellular release of K+. Such a mechanism would hardly influence the extracellular homeostasis in vitro due to the large medium-to-cell volume ratio. The findings demonstrate, nonetheless, the significance of a competent energy metabolism to support cell volume regulation. This is concluded from the failure of regulatory volume decrease of hypotonically suspended glial cells in anoxia plus iodoacetate.(ABSTRACT TRUNCATED AT 250 WORDS)


Genetic contribution of catechol-O-methyltransferase variants in treatment outcome of low back pain: a prospective genetic association study.
Treatment outcome of low back pain (LBP) is associated with inter-individual variations in pain relief and functional disability. Genetic variants of catechol-O-methyltransferase (COMT) gene have previously been shown to be associated with pain sensitivity and pain medication. This study examines the association between COMT polymorphisms and 7-11 year change in Oswestry Disability Index (ODI) and Visual Analog Score (VAS) for LBP as clinical outcome variables in patients treated with surgical instrumented lumbar fusion or cognitive intervention and exercise. 93 unrelated patients with chronic LBP for duration of >1 year and lumbar disc degeneration (LDD) were treated with lumbar fusion (N = 60) or cognitive therapy and exercises (N = 33). Standardised questionnaires assessing the ODI, VAS LBP, psychological factors and use of analgesics, were answered by patients both at baseline and at 7-11 years follow-up. Four SNPs in the COMT gene were successfully genotyped. Single marker as well as haplotype association with change in ODI and VAS LBP, were analyzed using Haploview, linear regression and R-package Haplostats. P-values were not formally corrected for multiple testing as this was an explorative study. Association analysis of individual SNPs adjusted for covariates revealed association of rs4633 and rs4680 with post treatment improvement in VAS LBP (p = 0.02, mean difference (β) = 13.5 and p = 0.02, β = 14.2 respectively). SNPs, rs4633 and rs4680 were found to be genotypically similar and in strong linkage disequilibrium (LD). A significant association was found with covariates, analgesics (p = 0.001, β = 18.6); anxiety and depression (p = 0.008, β = 15.4) and age (p = 0.03, mean difference per year (β) = 0.7) at follow-up. There was a tendency for better improvement among heterozygous patients compared to the homozygous. No association was observed for the analysis of the common haplotypes, these SNPs were situated on. Results suggest an influence of genetic variants of COMT gene in describing the variation in pain after treatment for low back pain. Replication in large samples with testing for other pain related genes is warranted.


Biological invasions as a component of global change in stressed marine ecosystems.
Biological invasions in marine environment are the lesser known aspect of global change. However, recent events which occurred in the Mediterranean Sea demonstrate that they represent a serious ecological and economical menace leading to biodiversity loss, ecosystem unbalancing, fishery and tourism impairment. In this paper we review marine bioinvasions using examples taken from the Mediterranean/Black Sea region. Particular attention is given to the environmental status of the receiving area as a fundamental pre-requisite for the colonisation success of alien species. The spread of the tropical algae belonging to the genus Caulerpa in the northwestern basin of the Mediterranean Sea has been facilitated by pre-existing conditions of instability of the Posidonia oceanica endemic ecosystem in relation to stress of both natural and anthropogenic origin. Human interventions caused long-term modification in the Black Sea environment, preparing a fertile ground for mass bioinvasion of aquatic nuisance species which, in some cases, altered the original equilibrium of the entire basin. Finally, the Venice lagoon is presented as the third example of an environment subjected to high propagule pressure and anthropogenic forcing and bearing the higher "diversity" of non-indigenous species compared to the other Mediterranean lagoons. Stressed environments are easily colonised by alien species; understanding the links between human and natural disturbance and massive development of non-indigenous species will help prevent marine bioinvasions, that are already favoured by global oceanic trade.


Autism and developmental disability caused by KCNQ3 gain-of-function variants.
Recent reports have described single individuals with neurodevelopmental disability (NDD) harboring heterozygous KCNQ3 de novo variants (DNVs). We sought to assess whether pathogenic variants in KCNQ3 cause NDD and to elucidate the associated phenotype and molecular mechanisms. Patients with NDD and KCNQ3 DNVs were identified through an international collaboration. Phenotypes were characterized by clinical assessment, review of charts, electroencephalographic (EEG) recordings, and parental interview. Functional consequences of variants were analyzed in vitro by patch-clamp recording. Eleven patients were assessed. They had recurrent heterozygous DNVs in KCNQ3 affecting residues R230 (R230C, R230H, R230S) and R227 (R227Q). All patients exhibited global developmental delay within the first 2 years of life. Most (8/11, 73%) were nonverbal or had a few words only. All patients had autistic features, and autism spectrum disorder (ASD) was diagnosed in 5 of 11 (45%). EEGs performed before 10 years of age revealed frequent sleep-activated multifocal epileptiform discharges in 8 of 11 (73%). For 6 of 9 (67%) recorded between 1.5 and 6 years of age, spikes became near-continuous during sleep. Interestingly, most patients (9/11, 82%) did not have seizures, and no patient had seizures in the neonatal period. Voltage-clamp recordings of the mutant KCNQ3 channels revealed gain-of-function (GoF) effects. Specific GoF variants in KCNQ3 cause NDD, ASD, and abundant sleep-activated spikes. This new phenotype contrasts both with self-limited neonatal epilepsy due to KCNQ3 partial loss of function, and with the neonatal or infantile onset epileptic encephalopathies due to KCNQ2 GoF. ANN NEUROL 2019;86:181-192.


Further insights on the inhibition of HIV type 1 infection in vitro by CD4-modified synthetic peptides containing phenylalanine.
Phenylalanine-containing peptides from CD4 were synthesized on the basis of chemical similarity with active CD4(81-92)-benzylated peptides. Systematic replacement of amino acids of these peptides bearing the benzyl group by phenylalanine, afforded several peptides that were able to block the binding of gp120 to CD4 and to inhibit HIV-induced syncytium formation. These experiments showed that substitution of residues 81 and 85 by phenylalanine was the most important for activity. Following optimization of the length of phenylalanine-substituted peptides it was found that FYICFVED and FYICFVEDE were the most active. Their IC50 for the inhibition of syncytium formation was around 1.2-1.6 microM. This activity is at least 30 times higher than that of the parent peptide FYIFFVEDQKEEDD previously reported (Lasarte et al., J Acquir Immune Defic Syndr 1994;7:129-134). Binding competition experiments with two different anti-peptide antisera recognizing the V3 region of gp120 and FYICFVEDE, show that the active peptides bind to V3 or to a sterically near region of V3. None of the active peptides was toxic to cells in vitro. The enhanced activity and simplicity of these new phenylalanine-substituted CD4 peptides might be a good starting point for the development of mimotopes of potential use for the treatment of AIDS.


Albendazole negatively regulates keratinocyte proliferation.
Increased keratinocyte proliferation occurs in the skin of psoriatic patients and is supposed to play a role in the pathogenesis of this disorder. Compounds interfering with keratinocyte proliferation could be useful in the management of psoriatic patients. To investigate whether albendazole, an anti-helmintic drug that regulates epithelial cell function in various systems, inhibits keratinocyte proliferation in models of psoriasis. Aldara-treated mice received daily topical application of albendazole. Keratinocyte proliferation and keratin (K) 6 and K16 expression were evaluated by immunohistochemistry and Western blotting and inflammatory cells/mediators were analysed by immunohistochemistry and real-time PCR. In human keratinocytes (HEKa and HaCaT) treated with albendazole, cell cycle and proliferation, keratins and cell cycle-associated factors were evaluated by flow cytometry, colorimetric assay and Western blotting respectively. Aldara-treated mice given albendazole exhibited reduced epidermal thickness, decreased number of proliferating keratinocytes and K6/K16 expression. Reduction of CD3- and Ly6G-positive cells in the skin of albendazole-treated mice associated with inhibition of IL-6, TNF-α, IL-1β, IL-17A, IL-36, CCL17, CXCL1, CXCL2 and CXCL5 expression. Treatment of keratinocytes with albendazole reduced K6/K16 expression and reversibly inhibited cell growth by promoting accumulation of cells in S-phase. This phenomenon was accompanied by down-regulation of CDC25A, a phosphatase regulating progression of cell cycle through S-phase, and PKR-dependent hyper-phosphorylation of eIF2α, an inhibitor of CDC25 translation. In Aldara-treated mice, albendazole activated PKR, enhanced eIF2α phosphorylation and reduced CDC25A expression. Data show that albendazole inhibits keratinocyte proliferation and exerts therapeutic effect in a murine model of psoriasis.


The tails of two geckos tell the story of dispersal in a fragmented landscape.
The fragmentation of habitat is a major cause of biodiversity loss. However, while numerous studies have suggested that reducing the size of populations and isolating them on fragments leads ultimately to the extinction of a species (small isolated populations are extinction prone), the evidence has been rather conjectural. This is because dispersal is so difficult to measure and isolation difficult to confirm. In past studies, evidence that populations become small and isolated on fragments, leading to declines, has relied on spatial patterns of distribution and abundance. Thus, a species not trapped in the matrix in which fragments are embedded might be assumed isolated on fragments, and if low in abundance on fragments compared to continuous habitat is assumed to have declined on fragments due to this isolation. However, without accurately measuring the degree of isolation, it is difficult to distinguish the role of isolation from other important causes of population decline that are correlated with fragment and population size, such as habitat degradation. Developments in molecular techniques and statistical methods now make it possible to measure isolation. Refreshingly, in this issue Hoehn et al. analyse microsatellite DNA with a suite of statistical methods to show convincingly that a declining species of gecko suffers from greater isolation on habitat fragments than a contrasting gecko that is able to disperse between fragments and hence persist in the severely fragmented wheatbelt of Western Australia.


Dermascope Use by Osteopathic Primary Care Physicians.
Osteopathic physicians (ie, DOs) in primary care may play a critical role in the early detection of skin cancer. Dermoscopy improves diagnostic accuracy; however, its use among primary care DOs remains understudied. To document dermascope use among DOs in primary care, to examine physician and practice characteristics associated with having used and having heard of a dermascope, and to examine the barriers to dermascope use. Osteopathic physicians were recruited to complete an anonymous survey assessing demographic factors, physician and practice characteristics, confidence in differentiating skin lesions, knowledge and dermascope use, and barriers to dermascope use. Bivariate analyses were conducted, and the significant factors were entered into 2 separate logistic regressions. A total of 768 participants were included in the study. Four hundred ten (54%) had heard of a dermascope, and 123 (15%) had used one. The statistically significant multivariate predictors for having used a dermascope (model 1) were graduating from medical school after 1989 and having greater confidence in differentiating skin lesions (OR, 2.2; 95% CI, 1.66-2.79). Those who graduated after 2009 were 9.5 times more likely and those graduating between 2000 and 2009 were 4.3 times more likely to have used a dermascope than those graduating before 1990 (95% CI, 4.29-20.90 and 95% CI, 2.04-9.23, respectively). Ever having heard of a dermascope (model 2) was associated with being female (OR, 1.4; 95% CI, 1.02-1.87); practicing in a group (OR, 1.6; 95% CI, 1.05-2.36), academic (OR, 2.2; 95% CI, 1.26-3.86), or community center (OR, 2.2; 95% CI, 1.20-4.00); and having greater confidence in differentiating skin lesions (OR, 1.3; 95% CI, 1.15-1.55). Both models were statistically significant and correctly classified 605 (84.1%) (model 1) and 444 (58.4%) participants (model 2). Dermascope use could help primary care DOs improve their diagnostic accuracy for skin cancer and reduce unnecessary referrals to specialists. Efforts to disseminate knowledge about the benefits of using a dermascope to primary care DOs are needed.


Gold nanorods decorated with a cancer drug for multimodal imaging and therapy.
Cancer, a condition with uncontrolled cell division, is the second leading cause of death worldwide. The currently available techniques for the imaging and treatment of cancer have their own limitations and hence a combination of more than one modality is expected to increase the efficacy of both diagnosis and treatment. In the present study, we have developed a multimodal imaging and therapeutic system by incorporating a chemotherapeutic drug, mitoxantrone (MTX) onto PEG coated gold nanorods (GNR). Strong absorption in the near-infrared (NIR) and visible regions qualifies GNR as an efficient photothermal (PTT) agent upon irradiation with either a NIR or visible laser. Additionally, the enhanced electric field of GNR makes it a suitable substrate for surface enhanced Raman scattering (SERS). Modification of GNR with amino PEG offers biocompatibility without affecting its optical property. In order to achieve tumor specificity, GNR-PEG was conjugated with tumor specific marker that can target cancer cells, leaving the normal cells unaffected. The incorporation of fluorescent chemotherapeutic drug mitoxantrone onto GNR-PEG facilitates chemotherapy as well as fluorescence imaging. The therapeutic efficacy of the developed GNR based system is tracked using fluorescence imaging and Raman imaging. The careful design of the system also facilitates the controlled release of the drug by photothermal triggering. Likewise, the imaging modality could be chosen as either Raman or fluorescence to monitor drug release in accordance with irradiation. The physico-chemical properties, and drug release profiles under different physiological conditions have been well studied. Finally, the developed system was tested for its therapeutic efficacy using cancer cells, in vitro. The receptor mediated cell uptake was more effective in folate receptor over-expressing cancer cells than in the normal and low-expressing cells. Accordingly the percentage of cell death was higher in folate receptor over-expressing cancer cells, which was further enhanced due to the effect of the dual therapeutic approach. The cell uptake and treatment efficacy was monitored using fluorescence microscopy and SERS. In conclusion, the developed GNR-PEG-MTX system is found to be an efficient multimodal therapeutic agent against cancer which could be tracked using two different techniques.


Structure and assembly of the influenza A virus ribonucleoprotein complex.
The genome of influenza A viruses consists of eight segments of single-stranded, negative-sense RNA that are encapsidated as individual rod-shaped ribonucleoprotein complexes (RNPs). Each RNP contains a viral RNA, a viral polymerase and multiple copies of the viral nucleoprotein (NP). Influenza A virus RNPs play important roles during virus infection by directing viral RNA replication and transcription, intracellular transport of the viral RNA, gene reassortment as well as viral genome packaging into progeny particles. As a unique genomic entity, the influenza A virus RNP has been extensively studied since the 1960s. Recently, exciting progress has been made in studying the RNP structure and its assembly, leading to a better understanding of the structural basis of various RNP functions.


Health and happiness: cross-sectional household surveys in Finland, Poland and Spain.
To explore the associations between health and how people evaluate and experience their lives. We analysed data from nationally-representative household surveys originally conducted in 2011-2012 in Finland, Poland and Spain. These surveys provided information on 10 800 adults, for whom experienced well-being was measured using the Day Reconstruction Method and evaluative well-being was measured with the Cantril Self-Anchoring Striving Scale. Health status was assessed by questions in eight domains including mobility and self-care. We used multiple linear regression, structural equation models and multiple indicators/multiple causes models to explore factors associated with experienced and evaluative well-being. The multiple indicator/multiple causes model conducted over the pooled sample showed that respondents with younger age (effect size, β = 0.19), with higher levels of education (β = -0.12), a history of depression (β = -0.17), poor health status (β = 0.29) or poor cognitive functioning (β = 0.09) reported worse experienced well-being. Additional factors associated with worse evaluative well-being were male sex (β = -0.03), not living with a partner (β = 0.07), and lower occupational (β = -0.07) or income levels (β = 0.08). Health status was the factor most strongly correlated with both experienced and evaluative well-being, even after controlling for a history of depression, age, income and other sociodemographic variables. Health status is an important correlate of well-being. Therefore, strategies to improve population health would also improve people's well-being.


NHS health checks through general practice: randomised trial of population cardiovascular risk reduction.
The global burden of the major vascular diseases is projected to rise and to remain the dominant non-communicable disease cluster well into the twenty first century. The Department of Health in England has developed the NHS Health Check service as a policy initiative to reduce population vascular disease risk. The aims of this study were to monitor population changes in cardiovascular disease (CVD) risk factors over the first year of the new service and to assess the value of tailored lifestyle support, including motivational interview with ongoing support and referral to other services. Randomised trial comparing NHS Health Check service only with NHS Health Check service plus additional lifestyle support in Stoke on Trent, England. Thirty eight general practices and 601 (365 usual care, 236 additional lifestyle support) patients were recruited and randomised independently between September 2009 and February 2010. Changes in population CVD risk between baseline and one year follow-up were compared, using intention-to-treat analysis. The primary outcome was the Framingham 10 year CVD risk score. Secondary outcomes included individual modifiable risk measures and prevalence of individual risk categories. Additional lifestyle support included referral to a lifestyle coach and free sessions as needed for: weight management, physical activity, cook and eat and positive thinking. Average population CVD risk decreased from 32.9% to 29.4% (p <0.001) in the NHS Health Check only group and from 31.9% to 29.2% (p <0.001) in the NHS Health Check plus additional lifestyle support group. There was no significant difference between the two groups at either measurement point. Prevalence of high blood pressure, high cholesterol and smoking were reduced significantly (p <0.01) in both groups. Prevalence of central obesity was reduced significantly (p <0.01) in the group receiving additional lifestyle support but not in the NHS Health Check only group. The NHS Health Check service in Stoke on Trent resulted in significant reduction in estimated population CVD risk. There was no evidence of further benefit of the additional lifestyle support services in terms of absolute CVD risk reduction.


Interventions for preventing obesity in children.
Prevention of childhood obesity is an international public health priority given the significant impact of obesity on acute and chronic diseases, general health, development and well-being. The international evidence base for strategies that governments, communities and families can implement to prevent obesity, and promote health, has been accumulating but remains unclear. This review primarily aims to update the previous Cochrane review of childhood obesity prevention research and determine the effectiveness of evaluated interventions intended to prevent obesity in children, assessed by change in Body Mass Index (BMI). Secondary aims were to examine the characteristics of the programs and strategies to answer the questions "What works for whom, why and for what cost?" The searches were re-run in CENTRAL, MEDLINE, EMBASE, PsychINFO and CINAHL in March 2010 and searched relevant websites. Non-English language papers were included and experts were contacted. The review includes data from childhood obesity prevention studies that used a controlled study design (with or without randomisation). Studies were included if they evaluated interventions, policies or programs in place for twelve weeks or more. If studies were randomized at a cluster level, six clusters were required. Two review authors independently extracted data and assessed the risk of bias of included studies. Data was extracted on intervention implementation, cost, equity and outcomes. Outcome measures were grouped according to whether they measured adiposity, physical activity (PA)-related behaviours or diet-related behaviours. Adverse outcomes were recorded. A meta-analysis was conducted using available BMI or standardized BMI (zBMI) score data with subgroup analysis by age group (0-5, 6-12, 13-18 years, corresponding to stages of developmental and childhood settings). This review includes 55 studies (an additional 36 studies found for this update). The majority of studies targeted children aged v 6-12 years. The meta-analysis included 37 studies of 27,946 children and demonstrated that programmes were effective at reducing adiposity, although not all individual interventions were effective, and there was a high level of observed heterogeneity (I2 = 82%). Overall, children in the intervention group had a standardised mean difference in adiposity (measured as BMI or zBMI) of -0.15 kg/m2 (95% confidence interval (CI): -0.21 to -0.09). Intervention effects by age subgroups were -0.26kg/m2 (95% CI -0.53 to 0.00) (0- 5 years), - 0.15 kg/m2 (95% CI -0.23 to -0.08) (6-12 years), and -0.09 kg/m2 (95% C I -0.20 to 0.03) (13-18 years). Heterogeneity was apparent in all three age groups and could not explained by randomisation status or the type, duration or setting of the intervention. Only eight studies reported on adverse effects and no evidence of adverse outcomes such as unhealthy dieting practices, increased prevalence of underweight or body image sensitivities was found. Interventions did not appear to increase health inequalities although this was examined in fewer studies. We found strong evidence to support beneficial effects of child obesity prevention programmes on BMI, particularly for programmes targeted to children aged six to 12 years. However, given the unexplained heterogeneity and the likelihood of small study bias, these findings must be interpreted cautiously. A broad range of programme components were used in these studies and whilst it is not possible to distinguish which of these components contributed most to the beneficial effects observed, our synthesis indicates the following to be promising policies and strategies: school curriculum that includes healthy eating, physical activity and body image; increased sessions for physical activity and the development of fundamental movement skills throughout the school week; improvements in nutritional quality of the food supply in schools; environments and cultural practices that support children eating healthier foods and being active throughout each day; support for teachers and other staff to implement health promotion strategies and activities (e.g. professional development, capacity building activities); parent support and home activities that encourage children to be more active, eat more nutritious foods and spend less time in screen based activities. However, study and evaluation designs need to be strengthened, and reporting extended to capture process and implementation factors, outcomes in relation to measures of equity, longer term outcomes, potential harms and costs. Childhood obesity prevention research must now move towards identifying how effective intervention components can be embedded within health, education and care systems and achieve long term sustainable impacts.


Genomic evidence of yellow fever virus in Aedes scapularis, southeastern Brazil, 2016.
The southeastern region of Brazil has recently experienced the largest yellow fever disease outbreak in decades. Since July 2016 epizootic events were reported in São Paulo state's north region, where 787 Culicidae were captured as part of public health surveillance efforts and tested using real-time quantitative PCR. One Aedes scapularis pool collected in November 2016 in an agriculture area in Urupês city tested positive for YFV-RNA. Using a validated multiplex PCR approach we were able to recover a complete virus genome sequence from this pool. Phylogenetic analysis of the novel strain and publicly available data indicates that the belongs to the South American genotype 1 clade circulating in Sao Paulo state and is basal to the recent outbreak clade in southeast Brazil. Our findings highlight the need of additional studies, including vector competence studies, to disentangle the role of Aedes scapularis in yellow fever transmission in the Americas.


Phosphorus transfer in runoff following application of fertilizer, manure, and sewage sludge.
Phosphorus (P) transfer in surface runoff from field plots receiving either no P, triplesuperphoshate (TSP), liquid cattle manure (LCS), liquid anaerobically digested sludge (LDS), or dewatered sludge cake (DSC) was compared over a 2-yr period. Dissolved inorganic P concentrations in runoff increased from 0.1 to 0.2 mg L(-1) on control and sludge-treated plots to 3.8 and 6.5 mg L(-1) following application of LCS and TSP, respectively, to a cereal crop in spring. When incorporated into the soil in autumn, runoff dissolved P concentrations were typically < 0.5 mg L(-1) across all plots, and particulate P remained the dominant P form. When surface-applied in autumn to a consolidated seedbed, direct loss of LCS and LDS increased both runoff volume and P transfers, but release of dissolved P occurred only from LCS. The largest P concentrations (>70 mg L(-1)) were recorded following TSP application without any increase in runoff volume, while application of bulky DSC significantly reduced total P transfers by 70% compared with the control due to a reduced runoff volume. Treatment effects in each monitoring period were most pronounced in the first runoff event. Differences in the release of P from the different P sources were related to the amounts of P extracted by either water or sodium bicarbonate in the order TSP > LCS > LDS > DSC. The results suggest there is a lower risk of P transfer in land runoff following application of sludge compared with other agricultural P amendments at similar P rates.


Variation in the ovarian reserve is linked to alterations in intrafollicular estradiol production and ovarian biomarkers of follicular differentiation and oocyte quality in cattle.
The mechanisms whereby the high variation in numbers of morphologically healthy oocytes and follicles in ovaries (ovarian reserve) may have an impact onovarian function, oocyte quality, and fertility are poorly understood. The objective was to determine whether previously validated biomarkers for follicular differentiation and function, as well as oocyte quality differed between cattle with low versus a high antral follicle count (AFC). Ovaries were removed (n = 5 per group) near the beginning of the nonovulatory follicular wave, before follicles could be identified via ultrasonography as being dominant, from heifers with high versus a low AFC. The F1, F2, and F3 follicles were dissected and diameters determined. Follicular fluid and thecal, granulosal, and cumulus cells and the oocyte were isolated and subjected to biomarker analyses. Although the size and numerous biomarkers of differentiation, such as mRNAs for the gonadotropin receptors, were similar, intrafollicular concentrations of estradiol and the abundance of mRNAs for CYP19A1 in granulosal cells and ESR1, ESR2, and CTSB in cumulus cells were greater, whereas mRNAs for AMH in granulosal cells and TBC1D1 in thecal cells were lower for animals with low versus a high AFC during follicle waves. Hence, variation in the ovarian reserve may have an impact on follicular function and oocyte quality via alterations in intrafollicular estradiol production and expression of key genes involved in follicle-stimulating hormone action (AMH) and estradiol (CYP19A1) production by granulosal cells, function and survival of thecal cells (TBC1D1), responsiveness of cumulus cells to estradiol (ESR1, ESR2), and cumulus cell determinants of oocyte quality (CTSB).


Cost-effectiveness of diagnostic algorithms including lateral-flow urine lipoarabinomannan for HIV-positive patients with symptoms of tuberculosis.
Tuberculosis (TB) is the leading cause of death among HIV-positive patients. We assessed the cost-effectiveness of including lateral-flow urine lipoarabinomannan (LF-LAM) in TB diagnostic algorithms for severely ill or immunosuppressed HIV-positive patients with symptoms of TB in Kenya. From a decision-analysis tree, ten diagnostic algorithms were elaborated and compared. All algorithms included clinical exam. The costs of each algorithm were calculated using a 'micro-costing' method. The efficacy was estimated through a prospective study that included severely ill or immunosuppressed (CD4<200cells/μL) HIV-positive adults with symptoms of TB. The cost-effectiveness analysis was performed using the disability-adjusted life year (DALY) averted as effectiveness outcome. A 4% discount rate was applied. The algorithm that added LF-LAM alone to the clinical exam lead to the least average cost per TB case detected (€47) and was the most cost-effective with a cost/DALY averted of €4.6. The algorithms including LF-LAM, microscopy and X-ray, and LF-LAM and Xpert in sputum, detected a high number of TB cases with a cost/DALY averted of €6.1 for each of them. In the comparisons of the algorithms two by two, using LF-LAM instead of microscopy (clinic&LAM vs clinic&microscopy) and using LF-LAM along with GeneXpert in sputum instead of GeneXpert in urine along with GeneXpert in sputum, (clinic&LAM&Xpert_sputum vs clinic&Xpert_sputum&Xpert_urine) led to the highest increase in the cost-effectiveness ratios (ICERs): €-7.2 and €-12.6 respectively. In these two comparisons, using LF-LAM increased the number of TB patients detected while reducing costs. Adding LF-LAM to smear microscopy alone or to smear microscopy and Xray led to the highest increase in the additional number of TB cases detected (31 and 25 respectively) with an incremental efficiency estimated at 134 and 344 DALYs respectively. The ICERs were €22.0 and €8.6 respectively. Including LF-LAM in TB diagnostic algorithms is cost-effective for severely ill or immunosuppressed HIV-positive patients.


Infrastructure Revisited: An Ethnographic Case Study of how Health Information Infrastructure Shapes and Constrains Technological Innovation.
Star defined infrastructure as something other things "run on"; it consists mainly of "boring things." Building on her classic 1999 paper, and acknowledging contemporary developments in technologies, services, and systems, we developed a new theorization of health information infrastructure with five defining characteristics: (1) a material scaffolding, backgrounded when working and foregrounded upon breakdown; (2) embedded, relational, and emergent; (3) collectively learned, known, and practiced (through technologically-supported cooperative work and organizational routines); (4) patchworked (incrementally built and fixed) and path-dependent (influenced by technical and socio-cultural legacies); and (5) institutionally supported and sustained (eg, embodying standards negotiated and overseen by regulatory and professional bodies). Our theoretical objective was, in a health care context, to explore what information infrastructure is and how it shapes, supports, and constrains technological innovation. Our empirical objective was to examine the challenges of implementing and scaling up video consultation services. In this naturalistic case study, we collected a total of 450 hours of ethnographic observations, over 100 interviews, and about 100 local and national documents over 54 months. Sensitized by the characteristics of infrastructure, we sought examples of infrastructural challenges that had slowed implementation and scale-up. We arranged data thematically to gain familiarity before undertaking an analysis informed by strong structuration, neo-institutional, and social practice theories, together with elements taken from the actor-network theory. We documented scale-up challenges at three different sites in our original case study, all of which relate to "boring things": the selection of a platform to support video-mediated consultations, the replacement of desktop computers with virtual desktop infrastructure profiles, and problems with call quality. In a fourth subcase, configuration issues with licensed video-conferencing software limited the spread of the innovation to another UK site. In all four subcases, several features of infrastructure were evident, including: (1) intricacy and lack of dependability of the installed base; (2) interdependencies of technologies, processes, and routines, such that a fix for one problem generated problems elsewhere in the system; (3) the inertia of established routines; (4) the constraining (and, occasionally, enabling) effect of legacy systems; and (5) delays and conflicts relating to clinical quality and safety standards. Innovators and change agents who wish to introduce new technologies in health services and systems should: (1) attend to materiality (eg, expect bugs and breakdowns, and prioritize basic dependability over advanced functionality); (2) take a systemic and relational view of technologies (versus as an isolated tool or function); (3) remember that technology-supported work is cooperative and embedded in organizational routines, which are further embedded in other routines; (4) innovate incrementally, taking account of technological and socio-cultural legacies; (5) consider standards but also where these standards come from and what priorities and interests they represent; and (6) seek to create leeway for these standards to be adapted to different local conditions.