Time-course and steroid specificity of aromatase induction in rat hypothalamus-preoptic area.
We studied the time-course and steroid specificity for aromatase induction in the hypothalamus-preoptic area (HPOA) of the adult male rat. Aromatase activity (AA) was measured in tissue homogenates by using a radiometric assay that quantifies the stereospecific production of 3H2O from [1 beta-3H] androstenedione. We found that by 48 h after administration of testosterone, HPOA AA was significantly (p less than 0.01) greater than control values in castrated rats. In contrast, AA was significantly (p less than 0.01) reduced 12 h after castration, and reached its lowest levels by 4 days after castration. Several other steroids, administered in 3-cm Silastic capsules for 7 days, were tested for their capacity to induce hypothalamic AA. In addition to testosterone, only 5 alpha-dihydrotestosterone and 5 alpha-androstane-3 alpha, 17 beta-diol were effective. Neither the stereoisomers of these compounds nor several other steroids, including estradiol, progesterone, and corticosterone, were active. This profile of activity indicates that the induction of HPOA AA is androgen-specific and, together with the demonstrated time-course of induction, lends further support to the hypothesis that androgens regulate AA through a receptor mechanism and the synthesis of new protein.


Clandestine ischemia in patients with vasospastic angina.
Coronary vasospasms generally occur at rest, but can also be triggered by physical exercise. Anginal pain and ST-segment elevation may be seen during exercise-stress tests. ST-segment depression, due to nonocclusive vasospasms, has also been found to occur. When the result of a test is positive, scintigraphy usually reveals perfusion defects. True silent or clandestine ischemia (normal result of exercise test with perfusion defects) in these patients is very uncommon. To stress the need for suspecting occurrence of coronary vasospasms in order to perform a proper diagnosis. Eight patients with angina were selected for this study. They had negative results of exercise tests with perfusion defects detected by thallium-201 tomography, normal coronary arteries and vasospasms. Maximal exercise-stress tests with thallium-201 tomography were performed. Sizes of perfusion defects were quantified by examining polar maps. Coronary angiography and then an intracoronary ergonovine test were performed for each patient. Significant defects were seen in territory of the right coronary artery, the left anterior descending artery, or both. Lung:heart ratio was normal in every case. The coronary arteries were normal and vasospasms were elicited with ergonovine in all the patients. Correspondence between the location of perfusion defects and angiographic spasms was generally observed. After treatment with calcium antagonists and nitrates all of them improved and defects detected by thallium tomography were no longer found when tests were repeated. Some patients with vasospastic angina may have normal results of exercise-stress tests and reversible perfusion defects detectable by scintigraphy. This finding must lead one to perform coronary angiography without administration of nitroglycerine beforehand and an ergonovine test if the coronary arteries are normal.


A role for the EphA family in the topographic targeting of vomeronasal axons.
We have investigated the role of the Eph family of receptor tyrosine kinases and their ligands in the establishment of the vomeronasal projection in the mouse. Our data show intriguing differential expression patterns of ephrin-A5 on vomeronasal axons and of EphA6 in the accessory olfactory bulb (AOB), such that axons with high ligand concentration project onto regions of the AOB with high receptor concentration and vice versa. These data suggest a mechanism for development of this projection that is the opposite of the repellent interaction between Eph receptors and ligands observed in other systems. In support of this idea, when given the choice of whether to grow on lanes containing EphA-F(c)/laminin or F(c)/laminin protein (in the stripe assay), vomeronasal axons prefer to grow on EphA-F(c)/laminin. Analysis of ephrin-A5 mutant mice revealed a disturbance of the topographic targeting of vomeronasal axons to the AOB. In summary, these data, which are derived from in vitro and in vivo experiments, indicate an important role of the EphA family in setting up the vomeronasal projection.


Comparison of cellular architecture, axonal growth, and blood vessel formation through cell-loaded polymer scaffolds in the transected rat spinal cord.
The use of multichannel polymer scaffolds in a complete spinal cord transection injury serves as a deconstructed model that allows for control of individual variables and direct observation of their effects on regeneration. In this study, scaffolds fabricated from positively charged oligo[poly(ethylene glycol)fumarate] (OPF(+)) hydrogel were implanted into rat spinal cords following T9 complete transection. OPF(+) scaffold channels were loaded with either syngeneic Schwann cells or mesenchymal stem cells derived from enhanced green fluorescent protein transgenic rats (eGFP-MSCs). Control scaffolds contained extracellular matrix only. The capacity of each scaffold type to influence the architecture of regenerated tissue after 4 weeks was examined by detailed immunohistochemistry and stereology. Astrocytosis was observed in a circumferential peripheral channel compartment. A structurally separate channel core contained scattered astrocytes, eGFP-MSCs, blood vessels, and regenerating axons. Cells double-staining with glial fibrillary acid protein (GFAP) and S-100 antibodies populated each scaffold type, demonstrating migration of an immature cell phenotype into the scaffold from the animal. eGFP-MSCs were distributed in close association with blood vessels. Axon regeneration was augmented by Schwann cell implantation, while eGFP-MSCs did not support axon growth. Methods of unbiased stereology provided physiologic estimates of blood vessel volume, length and surface area, mean vessel diameter, and cross-sectional area in each scaffold type. Schwann cell scaffolds had high numbers of small, densely packed vessels within the channels. eGFP-MSC scaffolds contained fewer, larger vessels. There was a positive linear correlation between axon counts and vessel length density, surface density, and volume fraction. Increased axon number also correlated with decreasing vessel diameter, implicating the importance of blood flow rate. Radial diffusion distances in vessels significantly correlated to axon number as a hyperbolic function, showing a need to engineer high numbers of small vessels in parallel to improving axonal densities. In conclusion, Schwann cells and eGFP-MSCs influenced the regenerating microenvironment with lasting effect on axonal and blood vessel growth. OPF(+) scaffolds in a complete transection model allowed for a detailed comparative, histologic analysis of the cellular architecture in response to each cell type and provided insight into physiologic characteristics that may support axon regeneration.


Yeast actin: polymerization kinetic studies of wild type and a poorly polymerizing mutant.
Wild-type actin and a mutant actin were isolated from yeast (Saccharomyces cerevisiae) and the polymerization properties were examined at pH 8.0 and 20 degrees C. The polymerization reaction was followed either by an increase in pyrene-labeled actin fluorescence or by a decrease in intrinsic fluorescence in the absence of pyrene-labeled actin. While similar to the properties of skeletal muscle actin, there are several important differences between the wild-type yeast and muscle actins. First, yeast actin polymerizes more rapidly than muscle actin under the same experimental conditions. The difference in rates may result from a difference in the steps involving formation of the nucleating species. Second, as measured with pyrene-labeled yeast actin, but not with intrinsic fluorescence, there is an overshoot in the fluorescence that has not been observed with skeletal muscle actin under the same conditions. Third, in order to simulate the polymerization process of wild-type yeast actin it is necessary to assume some fragmentation of the filaments. Finally, gelsolin inhibits polymerization of yeast actin but is known to accelerate the polymerization of muscle actin. A mutant actin (R177A/D179A) has also been isolated and studied. The mutations are at a region of contact between monomers across the long axis of the actin filament. This mutant polymerizes more slowly than wild type and filaments do not appear to fragment during polymerization. Elongation rates of the wild type and the mutant differ by only about 3-fold, and the slower polymerization of the mutant appears to result primarily from poorer nucleation.


Diversity study on Sclerotinia trifoliorum Erikks., the causal agent of clover rot in red clover crops (Trifolium pratense L.).
Since the 16th century, red clover has been an important crop in Europe. Since the 1940s, the European areal of red clover has been severely reduced, due to the availability of chemical fertilizers and the growing interest in maize. Nowadays there is a growing interest in red clover again, although some setbacks still remain. An important setback is the low persistence of red clover crops. Clover rot, caused by the ascomycete fungus Sclerotinia trifoliorum Erikss., is a major disease in Europe and reduces the persistence of red clover crops severely. The fungus infects clover plants through ascospores in the autumn, the disease develops during the winter and early spring and can kill many plants in this period. In early spring, black sclerotia, serving as surviving bodies, are formed on infected plants. Sclerotia can survive up to 7 years in the soil (Ohberg, 2006). The development of clover rot is highly dependent on the weather conditions: a humid fall, necessary for the germination of the ascospores and an overall warm winter with short periods of frost are favourable for the disease. Cold and dry winters slow the mycelial growth down too much and prevent the disease from spreading. Clover rot is difficult to control and completely resistant red clover varieties have yet to be developed. Because of the great annual variation in disease severity, plant breeders cannot use natural infection as an effective means to screen for resistant material. Breeding for resistant cultivars is being slowed down by the lack of a bio-test usable in breeding programs. When applying artificial infections, it is necessary to have an idea of the diversity of the pathogen. A diverse population will require resistance screening with multiple isolates. The objective of this research is to investigate the genetic diversity among isolates from the pathogen S. trifoliorum from various European countries. We assessed diversity using a species identification test based on the sequence of the beta-tubulin gene, vegetative compatibility grouping and AFLP.


OTX2 mutations contribute to the otocephaly-dysgnathia complex.
Otocephaly or dysgnathia complex is characterised by mandibular hypoplasia/agenesis, ear anomalies, microstomia, and microglossia; the molecular basis of this developmental defect is largely unknown in humans. This study reports a large family in which two cousins with micro/anophthalmia each gave birth to at least one child with otocephaly, suggesting a genetic relationship between anophthalmia and otocephaly. OTX2, a known microphthalmia locus, was screened in this family and a frameshifting mutation was found. The study subsequently identified in one unrelated otocephalic patient a sporadic OTX2 mutation. Because OTX2 mutations may not be sufficient to cause otocephaly, the study assayed the potential of otx2 to modify craniofacial phenotypes in the context of known otocephaly gene suppression in vivo. It was found that otx2 can interact genetically with pgap1, prrx1, and msx1 to exacerbate mandibular and midline defects during zebrafish development. However, sequencing of these loci in the OTX2-positive families did not unearth likely pathogenic lesions, suggesting further genetic heterogeneity and complexity. Identification of OTX2 involvement in otocephaly/dysgnathia in humans, even if loss of function mutations at this locus does not sufficiently explain the complex anatomical defects of these patients, suggests the requirement for a second genetic hit. Consistent with this notion, trans suppression of otx2 and other developmentally related genes recapitulate aspects of the otocephaly phenotype in zebrafish. This study highlights the combined utility of genetics and functional approaches to dissect both the regulatory pathways that govern craniofacial development and the genetics of this disease group.


Phytoene desaturase is localized exclusively in the chloroplast and up-regulated at the mRNA level during accumulation of secondary carotenoids in Haematococcus pluvialis (Volvocales, chlorophyceae).
The unicellular green alga Haematococcus pluvialis Flotow is known for its massive accumulation of ketocarotenoids under various stress conditions. Therefore, this microalga is one of the favored organisms for biotechnological production of these antioxidative compounds. Astaxanthin makes up the main part of the secondary carotenoids and is accumulated mostly in an esterified form in extraplastidic lipid vesicles. We have studied phytoene desaturase, an early enzyme of the carotenoid biosynthetic pathway. The increase in the phytoene desaturase protein levels that occurs following induction is accompanied by a corresponding increase of its mRNA during the accumulation period, indicating that phytoene desaturase is regulated at the mRNA level. We also investigated the localization of the enzyme by western-blot analysis of cell fractions and by immunogold labeling of ultrathin sections for electron microscopy. In spite of the fact that secondary carotenoids accumulate outside the chloroplast, no extra pathway specific for secondary carotenoid biosynthesis in H. pluvialis was found, at least at this early stage in the biosynthesis. A transport process of carotenoids from the site of biosynthesis (chloroplast) to the site of accumulation (cytoplasmatic located lipid vesicles) is implicated.


Intimate partner violence: associations with low infant birthweight in a South African birth cohort.
Violence against women is a global public health problem. Exposure to intimate partner violence (IPV) during pregnancy has been associated with a number of adverse maternal and fetal outcomes, including delivery of a low birthweight (LBW) infant. However, there is a paucity of data from low-middle income countries (LMIC). We examined the association between antenatal IPV and subsequent LBW in a South African birth cohort. This study reports data from the Drakenstein Child Lung Health Study (DCLHS), a multidisciplinary birth cohort investigation of the influence of a number of antecedent risk factors on maternal and infant health outcomes over time. Pregnant women seeking antenatal care were recruited at two different primary care clinics in a low income, semi-rural area outside Cape Town, South Africa. Antenatal trauma exposure was assessed using the Childhood Trauma Questionnaire (CTQ) and an IPV assessment tool specifically designed for the purposes of this study. Potential confounding variables including maternal sociodemographics, pregnancy intention, partner support, biomedical and mental illness, substance use and psychosocial risk were also assessed. Bivariate and multiple regression analyses were performed to determine the association between IPV during pregnancy and delivery of an infant with LBW and/or low weight-for-age z (WAZ) scores. The final study sample comprised 263 mother-infant dyads. In multiple regression analyses, the model run was significant [r2 = 0.14 (adjusted r2 = 0.11, F(8, 212) = 4.16, p = 0.0001]. Exposure to physical IPV occurring during the past year was found to be significantly associated with LBW [t = -2.04, p = 0.0429] when controlling for study site (clinic), maternal height, ethnicity, socioeconomic status, substance use and childhood trauma. A significant association with decreased WAZ scores was not demonstrated. Exposure of pregnant women to IPV may impact newborn health. Further research is needed in this field to assess the relevant underlying mechanisms, to inform public health policies and to develop appropriate trauma IPV interventions for LMIC settings.


Outcomes of primary percutaneous coronary intervention at a joint commission international accredited hospital in a developing country -- can good results, possibly similar to the west, be achieved?
Primary percutaneous coronary intervention (PCI) is the treatment of choice following ST-elevation myocardial infarction (STEMI). There is limited adoption and a paucity of data on outcomes following primary PCI in developing countries. The objective of this study was to describe the procedural and clinical outcomes of patients undergoing PCI for STEMI at a Joint Commission International Accreditation (JCIA) certified hospital in Pakistan and make a comparison with outcomes from the West. We conducted a retrospective cohort study at a tertiary care university hospital in Karachi, Pakistan. A total of 277 consecutive patients undergoing primary PCI between January 2001 and December 2005 were reviewed. Exclusion criteria included preceding fibrinolytic therapy and STEMI due to stent thrombosis. Cox proportional hazards models were constructed. The primary outcome was mortality. Procedural success was 97.1%. Inhospital mortality was 8.3% (43.9% in cardiogenic shock, 2.1% in non-shock patients), comparing very favorably with the published literature from developed countries. Multivariate predictors of death included (hazards ratio, 95% confidence interval) age (1.42 [1.14-1.76]), mechanical ventilation (8.35 [2.82-24.73]), cardiogenic shock (2.80 [1.04-7.55]), prior CABG (9.78 [1.15-83.13]) and ejection fraction (0.96 [0.92-0.99]). We conclude that excellent outcomes for a critical illness like STEMI can be achieved in a developing country at a JCIA-certified hospital, possibly similar to those seen in the West. There is a strong need for making the practice of primary PCI more widespread in developing nations. More outcomes data are needed from similar hospitals in the region to determine whether our results are generalizable.


Presynaptic regulation of isolated neonatal rat carotid body type I cells by histamine.
The type I cells are the chemoreceptive elements of the carotid bodies and are critical in defining the ventilatory response to hypoxia and hypercapnia. Recent evidence has suggested that histamine is released by the carotid body in response to hypoxia and acts as an excitatory neurotransmitter. Here we use isolated neonatal rat type I cells to assess the presynaptic actions of histamine and define the receptor subtypes that mediate them. All four histamine receptor subtypes are expressed on the type I cells, however activation of these receptors with histamine or selective agonists caused no rise in intracellular calcium ([Ca(2+)](i)) and histamine did not augment calcium entry or modulate macroscopic currents evoked in type I cells. Thus activation of histamine receptors on type I cells is unlikely to provide a presynaptic positive feedback mechanism during chemotransduction and any excitatory role attributed to the actions of histamine is likely to come from a postsynaptic effect on the carotid sinus nerve (CSN).


Acculturation, Skin Tone Preferences, and Tanning Behaviours Among Young Adult Asian Australians.
Australia has a significant proportion of residents of Asian heritage. Although the incidence of skin cancer is lower in those of Asian heritage than Caucasians, their prognosis is often worse. Sociocultural variables are central to the tanning behaviours of individuals from Western cultures. We examined the role of sociocultural variables in the tanning behaviours (outdoor tanning, indoor/solarium and fake tan use) among Asian Australians. A sample of 399 young adults identifying either as a person of Asian heritage or as Asian Australian participated in an online survey. Our results suggest that Asian Australians are at risk of skin cancer; over 35 % of the sample reported engaging in outdoor tanning and over 10 % in solarium tanning. After controlling for demographic factors and skin cancer knowledge, preferring a darker skin tone and being acculturated to Australia were significantly associated with tanning behaviour. Participants' low levels of skin cancer knowledge are of concern, and possibilities for improving knowledge levels in this group are considered. Further, we recommended that future research studies investigate sociocultural and appearance-related beliefs associated with tanning behaviours in this population, in order to determine best avenues for intervention.


ALK inhibitor PF02341066 (crizotinib) increases sensitivity to radiation in non-small cell lung cancer expressing EML4-ALK.
Crizotinib (PF02341066) is a tyrosine kinase inhibitor of anaplastic lymphoma kinase (ALK) that has been shown to selectively inhibit growth of cancer cells that harbor the EML4-ALK fusion found in a subset of patients with non-small cell lung cancer (NSCLC). While in clinical trials, PF02341066 has shown a significant therapeutic benefit as a single agent; the effectiveness of combining it with other therapeutic modalities including ionizing radiation remains unknown. To further elucidate the role of PF02341066 in tumor inhibition, we examined its effects alone and in combination with radiation on downstream signaling, apoptosis, and radiosensitivity in two NSCLC cell lines in vitro: H3122, which harbors the EML4-ALK fusion, and H460, which does not. We also examined the in vivo effects of PF02341066 in H3122 mouse xenografts. In the H3122 cell line, PF02341066 inhibited phosphorylation of ALK and its downstream effectors: AKT, ERK, and STAT3. H3122 cells treated with a combination of PF02341066 and radiation showed an increase in cellular apoptosis and were sensitized to radiation therapy (dose enhancement ratio, 1.43; P < 0.0001). Moreover, in an H3122 xenograft model, the combined treatment resulted in greater tumor growth inhibition than either treatment alone (P < 0.05). None of these effects was observed in the EML4-ALK-negative H460 cells. Our findings indicate that PF02341066 acts as a radiation sensitizer in cells harboring the EML4-ALK fusion, providing a rationale for a clinical trial combining ALK inhibitor with radiation in the NSCLCs expressing ALK.


Targeting deficient DNA damage repair in gastric cancer.
Over recent years our understanding of DNA damage repair has evolved leading to an expansion of therapies attempting to exploit DNA damage repair deficiencies across multiple solid tumours. Gastric cancer has been identified as a tumour where a subgroup of patients demonstrates deficiencies in the homologous recombination pathway providing a potential novel treatment approach for this poor prognosis disease. This review provides an overview of DNA damage repair and how this has been targeted to date in other tumour types exploiting the concept of synthetic lethality. This is followed by a discussion of how deficiencies in homologous recombination may be identified across tumour types and on recent progress in targeting DNA repair deficiencies in gastric cancer. Gastric cancer remains a difficult malignancy to treat and the possibility of targeting deficient DNA repair in a subgroup of patients is an exciting prospect. Future combinations with immunotherapy and radiotherapy are appealing and appear to have a sound biological rationale. However, much work remains to be done to understand the significance of the genetic and epigenetic alterations involved, to elucidate the optimum predictive signatures or biomarkers and to consider means of overcoming treatment resistance.


Hospitalisation and length of hospital stay following first-episode psychosis: systematic review and meta-analysis of longitudinal studies.
Reducing hospitalisation and length of stay (LOS) in hospital following first episode psychosis (FEP) is important, yet reliable measures of these outcomes and their moderators are lacking. We conducted a systematic review and meta-analysis to investigate the proportion of FEP cases who were hospitalised after their first contact with services and the LOS in a hospital during follow-up. Studies were identified from a systematic search across major electronic databases from inception to October 2017. Random effects meta-analyses and meta-regression analyses were conducted. 81 longitudinal studies encompassing data for 23 280 FEP patients with an average follow-up length of 7 years were included. 55% (95% CI 50.3-60.5%) of FEP cases were hospitalised at least once during follow-up with the pooled average LOS of 116.7 days (95% CI 95.1-138.3). Older age of illness onset and being in a stable relationship were associated with a lower proportion of people who were hospitalised. While the proportion of hospitalised patients has not decreased over time, LOS has, with the sharpest reduction in the latest time period. The proportion of patients hospitalised during follow-up was highest in Australia and New Zealand (78.4%) compared to Europe (58.1%) and North America (48.0%); and lowest in Asia (32.5%). Black ethnicity and longer duration of untreated psychosis were associated with longer LOS; while less severe psychotic symptoms at baseline were associated with shorter LOS. One in two FEP cases required hospitalisation at least once during a 7-year follow-up with an average length of hospitalisation of 4 months during this period. LOS has declined over time, particularly in those countries in which it was previously longest.


Antimicrobial, plant growth-promoting and genomic properties of the peanut endophyte Bacillus velezensis LDO2.
Peanut suffer from a number of fungal and bacterial pathogens, while plant endophytes were considered excellent candidates as biocontrol agents. In this study, the peanut endophytic bacterium LDO2 was evaluated for the potential of peanut pathogens inhibition and growth-promotion, and the genetic mechanisms were explored by genome mining. Strain LDO2 significantly inhibited the growth of peanut pathogenic fungi and pathogenic bacteria, and specifically, it showed pronounced inhibition on mycelia growth of Aspergillus flavus mycelia and caused mycelial deformity. Gene clusters responsible for antifungal metabolites (fengycin, surfactin, bacilysin) and antibacterial metabolites (butirosin, bacillaene, difficidin, macrolactin, surfactin, bacilysin) were identified. Strain LDO2 also exhibited several growth-promoting related features including phosphate solubilization, siderophore production and growth promotion of peanut root. Genes associated with plant growth promotion were also identified and analyzed, as well as genes related to secreted proteins. These findings suggested that this peanut endophyte could be a potential biocontrol agent in peanut production and a source of antimicrobial compounds for further exploitation.


Assembly of cell division proteins at the E. coli cell center.
Cell division in Escherichia coli requires the coordinated action of at least ten proteins. In recent years, substantial progress has been made in understanding the assembly of these proteins at the cell septum. These findings suggest a largely stepwise appearance of cell division proteins at the centre of the cell.


Non-myeloablative autologous haematopoietic stem cell transplantation expands regulatory cells and depletes IL-17 producing mucosal-associated invariant T cells in multiple sclerosis.
Autologous haematopoietic stem cell transplantation has been tried as one experimental strategy for the treatment of patients with aggressive multiple sclerosis refractory to other immunotherapies. The procedure is aimed at ablating and repopulating the immune repertoire by sequentially mobilizing and harvesting haematopoietic stem cells, administering an immunosuppressive conditioning regimen, and re-infusing the autologous haematopoietic cell product. 'Non-myeloablative' conditioning regimens to achieve lymphocytic ablation without marrow suppression have been proposed to improve safety and tolerability. One trial with non-myeloablative autologous haematopoietic stem cell transplantation reported clinical improvement and inflammatory stabilization in treated patients with highly active multiple sclerosis. The aim of the present study was to understand the changes in the reconstituted immune repertoire bearing potential relevance to its mode of action. Peripheral blood was obtained from 12 patients with multiple sclerosis participating in the aforementioned trial and longitudinally followed for 2 years. We examined the phenotype and function of peripheral blood lymphocytes by cell surface or intracellular staining and multi-colour fluorescence activated cell sorting alone or in combination with proliferation assays. During immune reconstitution post-transplantation we observed significant though transient increases in the proportion of CD4+ FoxP3+ T cells and CD56(high) natural killer cell subsets, which are cell subsets associated with immunoregulatory function. CD8+ CD57+ cytotoxic T cells were persistently increased after therapy and were able to suppress CD4+ T cell proliferation with variable potency. In contrast, a CD161(high) proinflammatory CD8+ T cell subset was depleted at all time-points post-transplantation. Phenotypic characterization revealed that the CD161(high)CD8+ T cells were mucosal-associated invariant T cells, a novel cell population originating in the gut mucosa but expressing the central nervous system-homing receptor CCR6. Detection of mucosal-associated invariant T cells in post-mortem multiple sclerosis brain white matter active lesions confirmed their involvement in the disease pathology. Intracellular cytokine staining demonstrated interferon γ and interleukin 17 production and lack of interleukin 10 production, a pro-inflammatory profile. Mucosal-associated invariant T cell frequency did not change in patients treated with interferon β; and was more depleted after autologous haematopoietic stem cell transplantation than in patients who had received high-dose cyclophosphamide (n = 7) or alemtuzumab (n = 21) treatment alone, suggesting an additive or synergistic effect of the conditioning regime components. We propose that a favourably modified balance of regulatory and pro-inflammatory lymphocytes underlies the suppression of central nervous system inflammation in patients with multiple sclerosis following non-myeloablative autologous haematopoietic stem cell transplantation with a conditioning regimen consisting of cyclophosphamide and alemtuzumab.


Age dependent electroencephalographic changes in attention-deficit/hyperactivity disorder (ADHD).
Objective biomarkers for attention-deficit/hyperactivity disorder (ADHD) could improve diagnostics or treatment monitoring of this psychiatric disorder. The resting electroencephalogram (EEG) provides non-invasive spectral markers of brain function and development. Their accuracy as ADHD markers is increasingly questioned but may improve with pattern classification. This study provides an integrated analysis of ADHD and developmental effects in children and adults using regression analysis and support vector machine classification of spectral resting (eyes-closed) EEG biomarkers in order to clarify their diagnostic value. ADHD effects on EEG strongly depend on age and frequency. We observed typical non-linear developmental decreases in delta and theta power for both ADHD and control groups. However, for ADHD adults we found a slowing in alpha frequency combined with a higher power in alpha-1 (8-10Hz) and beta (13-30Hz). Support vector machine classification of ADHD adults versus controls yielded a notable cross validated sensitivity of 67% and specificity of 83% using power and central frequency from all frequency bands. ADHD children were not classified convincingly with these markers. Resting state electrophysiology is altered in ADHD, and these electrophysiological impairments persist into adulthood. Spectral biomarkers may have both diagnostic and prognostic value.


Major transcriptome reprogramming underlies floral mimicry induced by the rust fungus Puccinia monoica in Boechera stricta.
Pucciniamonoica is a spectacular plant parasitic rust fungus that triggers the formation of flower-like structures (pseudoflowers) in its Brassicaceae host plant Boecherastricta. Pseudoflowers mimic in shape, color, nectar and scent co-occurring and unrelated flowers such as buttercups. They act to attract insects thereby aiding spore dispersal and sexual reproduction of the rust fungus. Although much ecological research has been performed on P. monoica-induced pseudoflowers, this system has yet to be investigated at the molecular or genomic level. To date, the molecular alterations underlying the development of pseudoflowers and the genes involved have not been described. To address this, we performed gene expression profiling to reveal 256 plant biological processes that are significantly altered in pseudoflowers. Among these biological processes, plant genes involved in cell fate specification, regulation of transcription, reproduction, floral organ development, anthocyanin (major floral pigments) and terpenoid biosynthesis (major floral volatile compounds) were down-regulated in pseudoflowers. In contrast, plant genes involved in shoot, cotyledon and leaf development, carbohydrate transport, wax biosynthesis, cutin transport and L-phenylalanine metabolism (pathway that results in phenylethanol and phenylacetaldehyde volatile production) were up-regulated. These findings point to an extensive reprogramming of host genes by the rust pathogen to induce floral mimicry. We also highlight 31 differentially regulated plant genes that are enriched in the biological processes mentioned above, and are potentially involved in the formation of pseudoflowers. This work illustrates the complex perturbations induced by rust pathogens in their host plants, and provides a starting point for understanding the molecular mechanisms of pathogen-induced floral mimicry.


Modeling radiation characteristics of semitransparent media containing bubbles or particles.
Modeling of radiation characteristics of semitransparent media containing particles or bubbles in the independent scattering limit is examined. The existing radiative properties models of a single particle in an absorbing medium using the approaches based on (1) the classical Mie theory neglecting absorption by the matrix, (2) the far field approximation, and (3) the near field approximation are reviewed. Comparison between models and experimental measurements are carried out not only for the radiation characteristics but also for hemispherical transmittance and reflectance of porous fused quartz. Large differences are found among the three models predicting the bubble radiative properties when the matrix is strongly absorbing and/or the bubbles are optically large. However, these disagreements are masked by the matrix absorption during calculation of radiation characteristics of the participating medium. It is shown that all three approaches can be used for radiative transfer calculations in an absorbing matrix containing bubbles.


Assessment of a membrane drinking water filter in an emergency setting.
The performance and acceptability of the Nerox(TM) membrane drinking water filter were evaluated among an internally displaced population in Pakistan. The membrane filter and a control ceramic candle filter were distributed to over 3,000 households. Following a 6-month period, 230 households were visited and filter performance and use were assessed. Only 6% of the visited households still had a functioning filter, and the removal performance ranged from 80 to 93%. High turbidity in source water (irrigation canals), together with high temperatures and large family size were likely to have contributed to poor performance and uptake of the filters.


Conditioned medium from human bone marrow-derived mesenchymal stem cells promotes skin moisturization and effacement of wrinkles in UVB-irradiated SKH-1 hairless mice.
Mesenchymal stem cells (MSCs) are promising therapeutic agents for various diseases. To investigate the effects of conditioned medium from human bone marrow-derived mesenchymal stem cells (MSC-CdM) on pro-collagen production and wrinkle formation, we performed in vitro and in vivo experiments. We assessed the effects of MSC-CdM on proliferation and photo-aging in human dermal fibroblasts after UVB exposure using enzyme activity assays for collagen type I secretion and MMP-1. To determine the effect of topically applied MSC-CdM on wrinkle formation, MSC-CdM (1% and 10%) and vehicle (propylene glycol: ethanol, 7 : 3) were applied to the dorsal skin of UVB-irradiated hairless mice for 8 weeks. We examined the effects on wrinkle formation by assessing visual skin grading, replica, tape stripping, transepidermal water loss (TEWL), and skin hydration measurement. We also examined histology of the lesions using hematoxylin-eosin, Masson's trichrome, and immunohistochemical staining. MSC-CdM markedly reduced UV-induced matrix metalloproteinase-1 expression and increased pro-collagen synthesis in a dose-dependent manner. Our findings suggest that MSC-CdM induces repair of dermal damage and effacement of wrinkles on UVB-irradiated hairless mice through protective effect of hydration. These results support an anti-wrinkle effect of MSC-CdM that involves increased collagen synthesis and suggest that MSC-CdM might be a potential candidate for preventing UV-induced skin damage.


Huntington disease.
Huntington disease is devastating to patients and their families - with autosomal dominant inheritance, onset typically in the prime of adult life, progressive course, and a combination of motor, cognitive and behavioural features. The disease is caused by an expanded CAG trinucleotide repeat (of variable length) in HTT, the gene that encodes the protein huntingtin. In mutation carriers, huntingtin is produced with abnormally long polyglutamine sequences that confer toxic gains of function and predispose the protein to fragmentation, resulting in neuronal dysfunction and death. In this Primer, we review the epidemiology of Huntington disease, noting that prevalence is higher than previously thought, geographically variable and increasing. We describe the relationship between CAG repeat length and clinical phenotype, as well as the concept of genetic modifiers of the disease. We discuss normal huntingtin protein function, evidence for differential toxicity of mutant huntingtin variants, theories of huntingtin aggregation and the many different mechanisms of Huntington disease pathogenesis. We describe the genetic and clinical diagnosis of the condition, its clinical assessment and the multidisciplinary management of symptoms, given the absence of effective disease-modifying therapies. We review past and present clinical trials and therapeutic strategies under investigation, including impending trials of targeted huntingtin-lowering drugs and the progress in development of biomarkers that will support the next generation of trials. For an illustrated summary of this Primer, visit: http://go.nature.com/hPMENh.


Can differences in the type, nature or amount of polysubstance use explain the increased risk of non-fatal overdose among psychologically distressed people who inject drugs?
This study investigates whether the type, nature or amount of polysubstance use can explain the increased risk of non-fatal overdose among people who inject drugs with severe psychological distress. Data came from three years (2011-2013) of the Illicit Drug Reporting System (IDRS), an annual sentinel sample of injecting drug users across Australia (n=2673). Structural Equation Modelling (SEM) was used on 14 drug types to construct five latent factors, each representing a type of polysubstance use. Tests of measurement invariance were carried out to determine if polysubstance use profiles differed between those with and without severe psychological distress. Next, we regressed non-fatal overdose on the polysubstance use factors with differences in the relationships tested between groups. Among those with severe psychological distress a polysubstance use profile characterised by heroin, oxycodone, crystal methamphetamine and cocaine use was associated with greater risk of non-fatal overdose. Among those without severe psychological distress, two polysubstance use profiles, largely characterised by opioid substitution therapies and prescription drugs, were protective against non-fatal overdose. The types of polysubstance use profiles did not differ between people who inject drugs with and without severe psychological distress. However, the nature of use of one particular polysubstance profile placed the former group at a strongly increased risk of non-fatal overdose, while the nature of polysubstance use involving opioid substitution therapies was protective only among the latter group. The findings identify polysubstance use profiles of importance to drug-related harms among individuals with psychological problems.


Randomized experiment on the effect of incentives and mailing strategy on response rates in a mail survey of dentists.
Response rates for mail surveys of dentists and other health care providers have declined appreciably over the past several decades. The objective of this study was to update evidence about the impact use of incentives and different mail strategies on response rates in a mail survey of dentists. We randomized private practice dentists in Iowa (N = 1267) into six study groups to test the effects of incentives and mail strategy on response rate. Survey incentives included either a $2 bill or a customized pen/stylus; a control group with no incentive was included. Mail strategies included bulk mail or first-class stamps. The overall survey response rate was 43%. A $2 bill incentive was associated with significantly higher likelihood of returning a survey compared to the control group with no incentive (48% response rate vs. 40%, respectively). A non-monetary incentive was not significantly associated with likelihood of response. Use of first-class stamps compared to bulk mail did not significantly affect response rate. This study demonstrated the effectiveness of a $2 incentive to increase response rate in a mail survey of dentists. Given the background trend of declining survey participation rates, researchers should consider use of monetary incentives and explore new strategies to increase participation among dentists and other health care providers.


The effect of bee pollen on secretion activity, markers of proliferation and apoptosis of porcine ovarian granulosa cells in vitro.
The general objective of this in vitro study was to examine the effect of bee pollen on the release of insulin-like growth factor I (IGF-I) and steroid hormone progesterone, and expression of markers of proliferation (PCNA) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF-I and progesterone were determined by RIA method and expression of PCNA and caspase-3 by immunocytochemistry. Bee pollen addition at the dose of 10 ng/mL significantly (P<0.05) inhibited IGF-I release by porcine ovarian granulosa cells. This growth factor was not influenced by 100 and 1000 ng/mL doses of bee pollen. Progesterone release by cells was not influenced by bee pollen addition at the doses of 10, 100 and 1000 ng/mL as used in our study. Similarly expression of PCNA and caspase-3 was not affected by bee pollen addition. The present study shows dose-dependent regulation of IGF-I by experimental bee pollen addition in vitro. Progesterone release, expression of PCNA and caspase-3 in porcine ovarian granulosa cells was not induced by pollen. Our results contribute to new insights regarding the possible effect of bee pollen on IGF-I release, which is important for regulation of porcine ovarian functions.


INITIAL EXPLORATION OF NEWLY IMPLEMENTED PUBLIC HEALTH POLICY USING GEOGRAPHIC INFORMATION SYSTEMS: THE CASE OF A U.S. SILVER ALERT PROGRAM.
Public health policies are designed for specific subsets of the population. Evidence that a policy is effectively designed should be based on whether it effectively addresses its mission. A critical factor is determining whether utilization patterns reflect the mission and the efficacy of public health policies, particularly during early stages of implementation. We assert that utilization patterns can be effectively assessed using geographic information systems (GIS). This paper uses the Silver Alert program, a recently implemented public health policy, as a case for how and why GIS can be used to examine utilization patterns. GIS are employed to visualize and spatially analyze a new health policy--North Carolina's Silver Alert policy. We use visualized data and spatial statistics to assess utilization patterns and mission adherence. Results show disproportionate utilization patterns of the Silver Alert policy. In particular, an outstanding number of Silver Alerts were used in Wake County and its surrounding counties, which are both the political and media center of North Carolina. Other counties, including populous counties, had few if any alerts. Findings suggest that the North Carolina's Silver Alert policy needs to be adjusted to more effectively address its mission. We identify several factors that need further examination prior to a statewide evaluation. From this case study, we propose ways future programs, particularly the introduction of the Affordable Care Act (ACA) in 2014, might use GIS to examine utilization patterns as a means to better understand whether and in what ways the health care needs of the public are being met with such a policy.


Day case arthroscopy under local anaesthesia.
A prospective study of the effectiveness of local anaesthesia in arthroscopy of the knee was performed in 212 consecutive patients. Arthroscopic surgery was undertaken successfully in 121 cases (57%), including meniscectomy and drilling of osteochondral defects. Dynamic evaluation of the patellofemoral joint articulation was possible and demonstration of pathological abnormalities was felt to be beneficial by some patients. The method described is safe, reliable, confers good postoperative analgesia and enables physiotherapy to begin immediately. Conversion to general anaesthesia was necessary in one case due to pain localised to a stiff and osteoarthritic hip. Intra-articular haemorrhage was found to be a problem in one case with synovitis. Only ten patients complained of moderate pain, none had severe pain. Local anaesthesia is contraindicated in cases with ipsilateral osteoarthritis of the hip or with significant synovitis of the knee. This technique is particularly suited to day case surgery.


Just Policy? An Ethical Analysis of Early Intervention Policy Guidance.
Early intervention (EI) aims to identify children or families at risk of poor health, and take preventative measures at an early stage, when intervention is more likely to succeed. EI is concerned with the just distribution of "life chances," so that all children are given fair opportunity to realize their potential and lead a good life; EI policy design, therefore, invokes ethical questions about the balance of responsibilities between the state, society, and individuals in addressing inequalities. We analyze a corpus of EI policy guidance to investigate explicit and implicit ethical arguments about who should be held morally responsible for safeguarding child health and well-being. We examine the implications of these claims and explore what it would mean to put the proposed policies into practice. We conclude with some remarks about the useful role that philosophical analysis can play in EI policy development.


A theoretical study on reductive debromination of polybrominated diphenyl ethers.
Recent progress has been made in the reductive debromination of polybrominated diphenyl ethers (PBDEs) by nanoscale zero-valent iron (nZVI). To better understand the mechanism of this reaction, seven selected BDE congeners and their anions were investigated at the density functional theory (DFT) level using four different methods, including B3LYP/6-31G(d), B3LYP/6-31+G(d), B3LYP/6-31G(d,p) and B3LYP/6-311G(d,p). The cleaved C-Br bonds observed in the equilibrium structures of anionic PBDEs were adopted as the probe of the susceptible debromination position of PBDEs in the presence of nZVI, and the proposed major reaction pathways based on our calculations can satisfactorily conform to the reported experimental results. The debromination preference is theoretically evaluated as meta-Br > ortho-Br > para-Br. In addition, both the calculated frontier orbital energies and adiabatic electronic affinities were found to be highly related to their experimental reductive debromination rate constants. The highest linear regression coefficient was observed in the case using the energy of lowest unoccupied molecular orbital as the molecular descriptor obtained from B3LYP/6-31G(d) (R(2) = 0.961, n = 7) or B3LYP/6-31G(d,p) (R(2) = 0.961, n = 7). The results clearly showed the evidence of an electron transfer mechanism associated with this reductive debromination reaction.


Importance of Superemitter Natural Gas Well Pads in the Marcellus Shale.
A large-scale study of methane emissions from well pads was conducted in the Marcellus shale (Pennsylvania), the largest producing natural gas shale play in the United States, to better identify the prevalence and characteristics of superemitters. Roughly 2100 measurements were taken from 673 unique unconventional well pads corresponding to ∼18% of the total population of active sites and ∼32% of the total statewide unconventional natural gas production. A log-normal distribution with a geometric mean of 2.0 kg h-1 and arithmetic mean of 5.5 kg h-1 was observed, which agrees with other independent observations in this region. The geometric standard deviation (4.4 kg h-1) compared well to other studies in the region, but the top 10% of emitters observed in this study contributed 77% of the total emissions, indicating an extremely skewed distribution. The integrated proportional loss of this representative sample was equal to 0.53% with a 95% confidence interval of 0.45-0.64% of the total production of the sites, which is greater than the U.S. Environmental Protection Agency inventory estimate (0.29%), but in the lower range of other mobile observations (0.09-3.3%). These results emphasize the need for a sufficiently large sample size when characterizing emissions distributions that contain superemitters.


Attitudes towards donor insemination--a post-Warnock survey.
In a multi-centred study, a total of 799 patients, donors and health-care professionals concerned with artificial insemination with donor semen (AID) responded to a questionnaire regarding their attitudes towards current provision of AID services and proposed legislation. There was little support for any fundamental change in the way in which AID is practised, at least in those centres. The anonymous status of the donor met with universal agreement. Although there was some support for the communication of non-identifying details to the recipient couple, where they wanted them, there was no support for any legislation which might give the AID child a right of access to details of the donor. The greatest divergence of opinion was over the question of who should have access to AID treatment and whether or not screening procedures should be applied to prospective parents. Most respondents felt that the closed and confidential relationship between the clinic and the other parties involved should not automatically be extended to general practitioners or any national bodies. In respect of specific recommendations of the Warnock Committee, there was support for changes which might legitimize or assist the present system, but not for any which might be restrictive.


Lymphoproliferative responses to synthetic peptides from merozoite ring-infected erythrocyte surface antigen and circumsporozoite protein: a longitudinal study during a falciparum malaria episode.
Proliferative responses of peripheral blood lymphocytes to synthetic peptides representing major epitopes of two malaria antigens (the merozoite ring-infected erythrocyte surface antigen and the sporozoite circumsporozoite protein) were investigated in Madagascar during a clinical Plasmodium falciparum episode. Thirty-seven patients greater than 10 years of age were enrolled at the beginning of the malaria transmission season and followed for four weeks. At enrollment, when the subjects presented with an acute infection, lymphocytes recovered from approximately 30% of them proliferated after peptide stimulation. These proliferative responses decreased sharply one and two weeks after treatment, with less than 10% responding to each peptide. Four weeks after treatment, the responses were only partially restored. The amplitude of these variations was not related to the initial parasitemia. At the individual level, proliferative response to each peptide varied greatly during the followup period, and this variation was unrelated to the presence of parasites in the blood.


Psychological mechanisms connected to dissociation: Generating hypotheses using network analyses.
A large number of mechanisms, many relating to the processing of affect, have been proposed to cause dissociation. The aim of this study was to use network analyses to identify psychological processes most closely connected with 'felt sense of anomaly' dissociative experiences. Both an undirected model and a partially directed network model were estimated using data from 6161 general population respondents collected online. The networks were used to identify relationships between dissociation and ten candidate mechanisms: cognitive appraisals, behavioural responses to dissociation, affect intolerance, alexithymia, attentional control, body vigilance, anxiety sensitivity, general self-efficacy, perseverative thinking, and beliefs regarding stress. Both models indicated a highly connected network in which dissociation had direct connections with six psychological processes: cognitive appraisals, behavioural responses, perseverative thinking, alexithymia, general self-efficacy, and beliefs about being overwhelmed. The strongest connection in both networks was between dissociation and cognitive appraisals (causal effect 0.73). The causal direction of connections could not be statistically determined with confidence, apart from the strong probability that dissociation causes meta-cognitions about being overwhelmed (98.54% of 50,000 sampled directed acyclic graphs). Both networks suggest that cognitive appraisals and factors relating to heightened (negative) sensitivity to affect are closely connected to dissociation. Dissociative experiences may arise from a high sensitivity to affect leading to threat-based appraisals that are ruminated upon and maintained by unhelpful behaviours such as avoidance. Investigation of these relationships in clinical groups, and direct causal tests, are required.


Use of concurrent multiple methods of contraception in the United States, 2008 to 2015.
To identify prevalence of, characteristics associated with, and combinations of, use of more than one method of contraception at last intercourse among US women between 2008 and 2015. We conducted bivariate and multivariable logistic regression analyses using data on concurrent contraceptive use from 2 nationally representative samples of women ages 15 to 44 who had used some form of contraception at last intercourse in the past 3 months in the 2006-2010 (n = 6601) and 2013-2017 (n = 5562) cycles of the National Survey of Family Growth. Use of more than one method of contraception at last sex increased from 14% in 2008 to 18% in 2015 (p<0.001), with increases in use documented across many population groups. Among multiple method users, the majority combined condoms with other methods (58%), while the rest combined other methods (42%). When compared to single method users, dual method users employing condoms are a more homogeneous group of individuals than are dual method users not employing condoms. As age increases, dual use with condoms decreases, but there is no similar linear relationship between age and dual method use without condoms. A sizable proportion of US women use more than one contraceptive method during sex; current estimates of contraceptive use focused exclusively on single method use may underestimate the extent to which women are protected from unintended pregnancy. The needs and goals of individuals combining contraceptive methods in different ways may change over the life course as pregnancy desires and life circumstances change. A sizable proportion of US women use more than one contraceptive method during sex; clinicians and health educators in nonclinical settings should assess and acknowledge these more complicated contraceptive strategies in order to help individuals achieve autonomy in method choice and meet their goals around pregnancy and sexually transmitted infection prevention.


Subjects with cardiovascular disease or high disease risk are more sedentary and less active than their healthy peers.
We investigated differences in objectively measured sedentary behaviour (SB) and physical activity (PA) levels in subjects with cardiovascular disease (CVD) diagnosis or high CVD risk compared with healthy controls. The present study includes a subsample (n=1398, Health 2011 Study) of participants, who attended health examinations and wore a triaxial accelerometer (≥4 days). Patients with CVD were identified and CVD risk was calculated for others using Framingham Risk Score (FRS). Participants were categorised into groups: FRS<10%; FRS=10%-30%; FRS>30%/CVD. Raw acceleration data were analysed with mean amplitude deviation (MAD) and angle for posture estimation (APE). MAD corresponding to intensity of PA was converted to metabolic equivalents (MET) and categorised to light (1.5-2.9 METs) and moderate to vigorous PA (MVPA≥3.0 METs). APE recognises SB and standing. Daily accumulated time of >30 s MVPA bouts was higher in FRS<10% group (46 min) than in FRS>30%/CVD group (29 min) (p<0.001). FRS>30%/CVD group were more sedentary, their mean daily number of >10 min SB bouts (13.2) was higher than in FRS <10% group (11.5) (p=0.002). Number and accumulated times of SB and PA bouts differed between the CVD risk groups. Causative research is required to assess the importance of SB and PA in prevention and rehabilitation of CVDs.


Essential fatty acid intake and serum fatty acid composition among adolescent girls in central Mozambique.
Many African diets are low in fat but are currently changing because of nutrition transition. We studied fat and fatty acid (FA) intake and the essential fatty acid (EFA) status of adolescent girls (aged 14-19 years, n 262) in Zambezia Province, central Mozambique. A cross-sectional study was carried out in a city as well as in the towns and rural villages of a coastal and an inland district. Dietary intake and FA sources were studied in a 24 h dietary recall. FA compositions of cholesteryl esters and phospholipids of non-fasting serum samples were analysed by GLC. Fat intake was low (13-18 % of energy) in all areas. Coconut and palm oil were the main sources of fat, and soyabean oil and maize were the main sources of PUFA. Compared to Food and Agriculture Organization/WHO 2010 recommendations, intake of linoleic acid (LA, 18 : 2n-6) was inadequate in the coastal district, and intakes of n-3 PUFA were inadequate in all areas. FA compositions of serum lipids differed between areas. The proportions of LA tended to be highest in the city and lowest in the rural areas. The phospholipid mead (20 : 3n-9):arachidonic acid (20 : 4n-6) ratio did not indicate EFA insufficiency. LA proportions in phospholipids were low, but those of long-chain n-6 and n-3 PUFA were high in comparison with Western adolescents. To conclude, fat sources, FA intake and EFA status differed between adolescent girls living in different types of communities. Fat intake was low, but EFA insufficiency was not indicated.


Clinical stakeholders' opinions on the use of selective decontamination of the digestive tract in critically ill patients in intensive care units: an international Delphi study.
Selective decontamination of the digestive tract (SDD) is a prophylactic antibiotic regimen that is not widely used in practice. We aimed to describe the opinions of key 'stakeholders' about the validity of the existing evidence base, likely consequences of implementation, relative importance of their opinions in influencing overall practice, likely barriers to implementation and perceptions of the requirement for further research to inform the decision about whether to embark on a further large randomised controlled trial. This was a Delphi study informed by comprehensive framework of possible determinants of health professionals' behaviour to study Critical Care practice in four countries. There were four key stakeholder participant groups including ICU physicians, pharmacists, clinical leads, and clinical microbiologists/ infectious disease physicians. Round one comprised participant interviews and Rounds two and three were online questionnaires using Delphi method. In this study, 141 participants were recruited of whom 82% were retained. Participants rated themselves as knowledgeable about SDD. Antibiotic resistance was identified as the most important issue. SDD was seen as a low clinical priority but few participants reported strong opposition. There was moderate agreement that research to date has not adequately addressed concerns about antibiotic resistance and lacks generalizability. Participants indicated equipoise with regard to benefits and harms of SDD, and indicated strong support for a further randomised trial. Clinicians have clinical equipoise about the effectiveness of SDD. Future research requires longer follow up to assess antibiotic resistance as well as greater validity/generalizability to provide definitive answers on the effectiveness of decontamination and effects on antibiotic resistance. SDD was regarded as not being a high clinical priority, which may limit future trial participation. These results have identified that further large randomised controlled trial of SDD in critical care is both warranted and appropriate.


Over-expression of TaDWF4 increases wheat productivity under low and sufficient nitrogen through enhanced carbon assimilation.
There is a strong pressure to reduce nitrogen (N) fertilizer inputs while maintaining or increasing current cereal crop yields. We show that overexpression of TaDWF4-B, the dominant shoot expressed homoeologue of OsDWF4, in wheat can increase plant productivity by up to 105% under a range of N levels on marginal soils, resulting in increased N use efficiency (NUE). We show that a two to four-fold increase in TaDWF4 transcript levels enhances the responsiveness of genes regulated by N. The productivity increases seen were primarily due to the maintenance of photosystem II operating efficiency and carbon assimilation in plants when grown under limiting N conditions and not an overall increase in photosynthesis capacity. The increased biomass production and yield per plant in TaDWF4 OE lines could be linked to modified carbon partitioning and changes in expression pattern of the growth regulator Target Of Rapamycin, offering a route towards breeding for sustained yield and lower N inputs.


Progression of brain cholinergic dysfunction in patients with isolated rapid eye movement sleep behavior disorder.
Reduced cortical acetylcholinesterase activity, as measured by 11 C-donepezil positron emission tomography (PET), has been reported in patients with isolated rapid eye movement (REM) sleep behavior disorder (iRBD). However, its progression and clinical implications have not been fully investigated. Here, we explored the relationship between longitudinal changes in brain acetylcholinesterase activity and cognitive function in iRBD. Twelve iRBD patients underwent 11 C-donepezil PET at baseline and after 3 years. PET images were interrogated with statistical parametric mapping (SPM) and a regions of interest (ROI) approach. Clinical progression was assessed with the Movement Disorder Society-Unified Parkinson's Disease Rating Scale-Part III (MDS-UPDRS-III). Cognitive function was rated using the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA). From baseline to follow-up, the mean 11 C-donepezil distribution volume ratio (DVR) decreased in the cortex (p = 0.006), thalamus (p = 0.013), and caudate (p = 0.013) ROI. Despite no significant changes in the group mean MMSE or MoCA scores being observed, individually, seven patients showed a decline in their scores on these cognitive tests. Subgroup analysis showed that only the subgroup of patients with a decline in cognitive scores had a significant reduction in mean cortical 11 C-donepezil DVR. Our results show that severity of brain cholinergic dysfunction in iRBD patients increases significantly over 3 years, and those changes are more severe in those with a decline in cognitive test scores.


Role of Phosphodiesterase-5 Inhibitors in Acute Right Ventricular Failure Due to Pulmonary Embolism.
BACKGROUND Sildenafil is a pulmonary vasodilator and its efficacy has been well established in patients with group 1 pulmonary hypertension. There are no established guidelines regarding its use in acute right ventricle failure. In our experience, it can be used as an adjunctive therapy in acute right ventricle failure due to pulmonary embolism, to reduce right ventricle afterload and hence improve size and function of the right ventricle. CASE REPORT This is a case report where sildenafil was used as a rescue agent to achieve improvement in the right ventricle size and function in a case of acute onset massive pulmonary embolism with acute right ventricle failure in the scenario where systemic thrombolytic therapy was contraindicated. CONCLUSIONS Improvement of right ventricle size and function was achieved using phosphodiesterase-5 Inhibitors in a case of acute right ventricle failure due to acute massive pulmonary embolism. There are no established guidelines regarding this clinical approach, however, given its efficacy in this case as adjunctive therapy in treatment of acute right ventricle, larger studies are needed to further establish its utility.


Novel and recurrent TRPV4 mutations and their association with distinct phenotypes within the TRPV4 dysplasia family.
Mutations in TRPV4, a gene that encodes a Ca(2+) permeable non-selective cation channel, have recently been found in a spectrum of skeletal dysplasias that includes brachyolmia, spondylometaphyseal dysplasia, Kozlowski type (SMDK) and metatropic dysplasia (MD). Only a total of seven missense mutations were detected, however. The full spectrum of TRPV4 mutations and their phenotypes remained unclear. To examine TRPV4 mutation spectrum and phenotype-genotype association, we searched for TRPV4 mutations by PCR-direct sequencing from genomic DNA in 22 MD and 20 SMDK probands. TRPV4 mutations were found in all but one MD subject. In total, 19 different heterozygous mutations were identified in 41 subjects; two were recurrent and 17 were novel. In MD, a recurrent P799L mutation was identified in nine subjects, as well as 10 novel mutations including F471del, the first deletion mutation of TRPV4. In SMDK, a recurrent R594H mutation was identified in 12 subjects and seven novel mutations. An association between the position of mutations and the disease phenotype was also observed. Thus, P799 in exon 15 is a hot codon for MD mutations, as four different amino acid substitutions have been observed at this codon; while R594 in exon 11 is a hotspot for SMDK mutations. The TRPV4 mutation spectrum in MD and SMDK, which showed genotype-phenotype correlation and potential functional significance of mutations that are non-randomly distributed over the gene, was presented in this study. The results would help diagnostic laboratories establish efficient screening strategies for genetic diagnosis of the TRPV4 dysplasia family diseases.


The Arabidopsis Protein Phosphatase PP2C38 Negatively Regulates the Central Immune Kinase BIK1.
Plants recognize pathogen-associated molecular patterns (PAMPs) via cell surface-localized pattern recognition receptors (PRRs), leading to PRR-triggered immunity (PTI). The Arabidopsis cytoplasmic kinase BIK1 is a downstream substrate of several PRR complexes. How plant PTI is negatively regulated is not fully understood. Here, we identify the protein phosphatase PP2C38 as a negative regulator of BIK1 activity and BIK1-mediated immunity. PP2C38 dynamically associates with BIK1, as well as with the PRRs FLS2 and EFR, but not with the co-receptor BAK1. PP2C38 regulates PAMP-induced BIK1 phosphorylation and impairs the phosphorylation of the NADPH oxidase RBOHD by BIK1, leading to reduced oxidative burst and stomatal immunity. Upon PAMP perception, PP2C38 is phosphorylated on serine 77 and dissociates from the FLS2/EFR-BIK1 complexes, enabling full BIK1 activation. Together with our recent work on the control of BIK1 turnover, this study reveals another important regulatory mechanism of this central immune component.


Extensive organohalogen contamination in wildlife from a site in the Yangtze River Delta.
The environmental and human health concerns for organohalogen contaminants (OHCs) extend beyond the 23 persistent organic pollutants (POPs) regulated by the Stockholm Convention. The current, intense industrial production and use of chemicals in China and their bioaccumulation makes Chinese wildlife highly suitable for the assessment of legacy, novel and emerging environmental pollutants. In the present study, six species of amphibians, fish and birds were sampled from paddy fields in the Yangtze River Delta (YRD) were screened for OHCs. Some extensive contamination was found, both regarding number and concentrations of the analytes, among the species assessed. High concentrations of chlorinated paraffins were found in the snake, Short-tailed mamushi (range of 200-340 μg g(-)(1)lw), Peregrine falcon (8-59 μg g(-1)lw) and Asiatic toad (97 μg g(-)(1)lw). Novel contaminants and patterns were observed; octaCBs to decaCB made up 20% of the total polychlorinated biphenyls (PCBs) content in the samples and new OHCs, substituted with 5-8 chlorines, were found but are not yet structurally confirmed. In addition, Dechlorane 602 (DDC-DBF) and numerous other OHCs (DDTs, hexachlorocyclohexanes (HCHs), polybrominated diphenyl ethers (PBDEs), hexbromocyclododecane (HBCDD), chlordane, heptachlor, endosulfan and Mirex) were found in all species analyzed. These data show extensive chemical contamination of wildlife in the YRD with a suite of OHCs with both known and unknown toxicities, calling for further in-depth studies.


Impedance control is selectively tuned to multiple directions of movement.
Humans are able to learn tool-handling tasks, such as carving, demonstrating their competency to make movements in unstable environments with varied directions. When faced with a single direction of instability, humans learn to selectively co-contract their arm muscles tuning the mechanical stiffness of the limb end point to stabilize movements. This study examines, for the first time, subjects simultaneously adapting to two distinct directions of instability, a situation that may typically occur when using tools. Subjects learned to perform reaching movements in two directions, each of which had lateral instability requiring control of impedance. The subjects were able to adapt to these unstable interactions and switch between movements in the two directions; they did so by learning to selectively control the end-point stiffness counteracting the environmental instability without superfluous stiffness in other directions. This finding demonstrates that the central nervous system can simultaneously tune the mechanical impedance of the limbs to multiple movements by learning movement-specific solutions. Furthermore, it suggests that the impedance controller learns as a function of the state of the arm rather than a general strategy.


An On-the-Fly Surface-Hopping Program JADE for Nonadiabatic Molecular Dynamics of Polyatomic Systems: Implementation and Applications.
Nonadiabatic dynamics simulations have rapidly become an indispensable tool for understanding ultrafast photochemical processes in complex systems. Here, we present our recently developed on-the-fly nonadiabatic dynamics package, JADE, which allows researchers to perform nonadiabatic excited-state dynamics simulations of polyatomic systems at an all-atomic level. The nonadiabatic dynamics is based on Tully's surface-hopping approach. Currently, several electronic structure methods (CIS, TDHF, TDDFT(RPA/TDA), and ADC(2)) are supported, especially TDDFT, aiming at performing nonadiabatic dynamics on medium- to large-sized molecules. The JADE package has been interfaced with several quantum chemistry codes, including Turbomole, Gaussian, and Gamess (US). To consider environmental effects, the Langevin dynamics was introduced as an easy-to-use scheme into the standard surface-hopping dynamics. The JADE package is mainly written in Fortran for greater numerical performance and Python for flexible interface construction, with the intent of providing open-source, easy-to-use, well-modularized, and intuitive software in the field of simulations of photochemical and photophysical processes. To illustrate the possible applications of the JADE package, we present a few applications of excited-state dynamics for various polyatomic systems, such as the methaniminium cation, fullerene (C20), p-dimethylaminobenzonitrile (DMABN) and its primary amino derivative aminobenzonitrile (ABN), and 10-hydroxybenzo[h]quinoline (10-HBQ).


The role of spinal angiography in the evaluation and treatment of pediatric spinal vascular pathology: a case series and systematic review.
While it has been extensively studied in adults, the role of spinal angiography in children with suspected spinal vascular malformations is not fully characterized. With special implications regarding technique, radiation dose, and pathology, we sought to review our single-center experience with pediatric spinal vascular pathology and use a systematic review of the literature to further identify its role in pediatrics. A retrospective chart review was conducted under IRB approval for all patients age 0-18 years old who underwent spinal angiography at our institution between 2007 and 2018 for concern for spinal vascular pathology. A simultaneous systematic review was conducted via dedicated search terms in two distinct databases and reviewed to identify all studies referring to spinal angiography or angiograms in pediatric patients. Six patients were included. Three patients (50%) had vascular malformations confirmed on diagnostic angiography and underwent subsequent endovascular treatment. Two patients (33.3%), one with hematomyelia and one with spinal cord infarction, had negative diagnostic angiograms. One patient (16.7%) had a spinal tumor and underwent angiography for further evaluation preoperatively. Spinal angiography was used to aid in diagnosis, preoperative planning, and treatment in these cases. It was demonstrated to be safe in this patient population, with no untoward events, minimal radiation dose, and possible therapeutic applications in select cases. The systematic review identified 11 studies regarding pediatric spinal angiography. These ranged from single case reports to case series of up to 38 patients and highlighted the role of spinal angiography in diagnosis, endovascular treatment, preoperative planning, and postoperative follow-up. Spinal angiography may be used in a variety of scenarios to better understand the architecture of vascular spinal lesions and facilitate endovascular intervention. While its application in both adult and pediatric pathology is limited to select cases, spinal angiography remains a key diagnostic procedure when approaching vascular lesions or tumors of the spine, assessing for etiology of spinal cord infarcts, and in the evaluation of unexplained hemorrhage in the spinal canal.


Patient-centred measurement in ophthalmology--a paradigm shift.
Ophthalmologists and researchers in ophthalmology understand what a rapidly evolving field ophthalmology is, and that to conduct good research it is essential to use the latest and best methods. In outcomes research, one modern initiative has been to conduct holistic measurement of outcomes inclusive of the patient's point of view; patient-centred outcome. This, of course, means including a questionnaire. However, the irony of trying to improve outcomes research by being inclusive of many measures is that the researcher may not be expert in all measures used. Certainly, few people conducting outcomes research in ophthalmology would claim to be questionnaire experts. Most tend to be experts in their ophthalmic subspecialty and probably simply choose a popular questionnaire that appears to fit their needs and think little more about it. Perhaps, unlike our own field, we assume that the field of questionnaire research is relatively stable. This is far from the case. The measurement of patient-centred outcomes with questionnaires is a rapidly evolving field. Indeed, over the last few years a paradigm shift has occurred in patient-centred measurement.


Differential transport properties of two mdr gene products are distinguished by progesterone.
P-glycoprotein is an integral membrane protein that is overproduced in multidrug-resistant cells. It is likely to function as an energy-dependent drug efflux pump to maintain intracellular drug concentrations below cytotoxic levels. Individually isolated multidrug-resistant murine cell lines, J7.V1-1 and J7.V3-1, overproduce P-glycoproteins encoded by the mdr1b and mdr1a genes, respectively. The transport properties of these cell lines and the drug binding characteristics of their P-glycoproteins have been compared. It is concluded that 1) the mdr1a gene product is a more efficient efflux pump than the mdr1b gene product, and 2) whereas a single class of vinblastine binding sites is present in J7.V1-1 membrane vesicles, there appears to be two classes of such sites in J7.V3-1 membrane vesicles. The effects of verapamil and progesterone, two compounds that are known to interact with P-glycoprotein, have been analyzed in the two cell lines. Progesterone inhibited drug binding and efflux and increased drug sensitivity to vinblastine with more potency in J7.V1-1 cells than in J7.V3-1 cells. It is concluded that progesterone, but not verapamil, can be used to differentiate the two mdr gene products in the mouse.


Effect of p-chloroamphetamine on 5-HT1A and 5-HT7 serotonin receptor expression in rat brain.
The aim of this study was to investigate if p-chloroamphetamine (PCA), which is neurotoxic to serotonin (5-HT) nerve terminals, was able to induce, like 3,4-methylenedioxymethamphetamine, a region-specific regulation of 5-HT1A receptor mRNA expression. The effect of PCA on the expression of 5-HT7 receptors, which share some pharmacological properties with 5-HT1A receptors, was comparatively studied. PCA (2 x 5 mg/kg) produced a lasting depletion of 5-HT content in the rat frontal cortex and hippocampus. In the hippocampus, the maximal 5-HT depletion was found on day 21 (-70%), whereas in the cortex, the highest 5-HT depletion was found on day 14 (-73%), with a partial but significant recovery on day 21. At the latter time point, 5-HT1A receptor mRNA expression was increased by 80% in the cortex and decreased by 50% in the hippocampus. The 5-HT1A receptor mRNA expression was also enhanced after exposure to PCA of rat cortical but not of hippocampal primary cultures. In regard to 5-HT7 receptor mRNA expression, the most remarkable change after PCA was the great increase (+200%) in the brain-stem. Binding studies to 5-HT1A receptors matched the changes in receptor mRNA expression. Gel shift assays revealed enhanced nuclear protein binding to the KB sequence with use of cortical but not hippocampal extracts of PCA-treated rats. Overall, the data show region-specific changes in 5-HT receptor-type expression that may not be entirely dependent on the neurotoxic effect of PCA on 5-HT terminals.


HP1β is a biomarker for breast cancer prognosis and PARP inhibitor therapy.
Members of the heterochromatin protein 1 family (HP1α, β and γ) are mostly associated with heterochromatin and play important roles in gene regulation and DNA damage response. Altered expression of individual HP1 subtype has profound impacts on cell proliferation and tumorigenesis. We analyzed the expression profile of HP1 family by data mining using a published microarray data set coupled with retrospective immunohistochemistry analyses of archived breast cancer biospecimens. We found that the patient group overexpressing HP1β mRNA is associated with poorly differentiated breast tumors and with a significantly lower survival rate. Immunohistochemical staining against HP1α, HP1β and HP1γ shows that respective HP1 expression level is frequently altered in breast cancers. 57.4-60.1% of samples examined showed high HP1β expression and 39.9-42.6 % of examined tumors showed no or low expression of each HP1 subtype. Interestingly, comparative analysis on HP1 expression profile and breast cancer markers revealed a positive correlation between the respective expression level of all three HP1 subtypes and Ki-67, a cell proliferation and well-known breast cancer marker. To explore the effect of individual HP1 on PARP inhibitor therapy for breast cancer, MCF7 breast cancer cells and individually HP1-depleted MCF7 cells were treated with PARP inhibitor ABT-888 with or without carboplatin. Notably, HP1β-knockdown cells are hypersensitive to the PARP inhibitor ABT-888 alone and its combination with carboplatin. In summary, while increased HP1β expression is associated with the poor prognosis in breast cancer, compromised HP1β abundance may serve as a useful predictive marker for chemotherapy, including PARP inhibitors against breast cancer.


The challenge of choosing environmental indicators of anthropogenic impacts in estuaries.
Ecological assessments over large spatial scales require that anthropogenic impacts be distinguishable above natural variation, and that monitoring tools are implemented to maximise impact detection and minimise cost. For three heavily modified and four relatively 'pristine' estuaries (disturbance category), chemical indicators (metals and PAHs) of anthropogenic stress were measured in benthic sediments, suspended sediments and deployed oysters, together with other environmental variables. These were compared with infaunal and hard-substrate invertebrate communities. Univariate analyses were useful for comparing contaminant loads between different monitoring tools and identified the strongest relationships between benthic and suspended sediments. However, multivariate analyses were necessary to distinguish ecological response to anthropogenic stressors from environmental "noise" over a large spatial scale and to identify sites that were being impacted by contaminants. These analyses provide evidence that suspended sediments are a useful alternative monitoring tool to detect potential anthropogenic impacts on benthic (infaunal and hard-substrate) communities.


Bilateral Achilles tendon ossification: diagnosis with ultrasonography and Single Photon Emission Computed Tomography/Computed Tomography. Case report.
Ossification of the Achilles tendon is a rare clinical condition characterized by the presence of one or more segments of variable sized ossified mass within the tendon. The cause of the condition is obscure. We present the case of a 41 year-old male patient with pain in the Achilles tendon due to bilateral ossification of the tendon near the calcaneus insertion. The case was evaluated by ultrasonography, single photon emission computed tomography, and transmission computed tomography. In this case, we regarded repetitive microtrauma as the cause of the ossification based on the bilateral occurrence.


Quantitative Association Between Peripapillary Bruch's Membrane Shape and Intracranial Pressure.
The purpose of this study was to determine if there is a quantitative relationship between chronic intracranial pressure (ICP) and peripapillary Bruch's membrane (pp-BM) shape and to determine whether change in pp-BM shape can be detected within 1 hour after ICP lowering by lumbar puncture (LP). In this study, 30° nasal-temporal optical coherence tomography B-scans were obtained within 1 hour before and after LP in 39 eyes from 20 patients (age = 23-86 years, 75% female, ICP [opening pressure] = 10-55 cm H2O). A total of 16 semi-landmarks defined pp-BM on each image. Geometric morphometric analysis identified principal components of shape in the image set. Generalized estimating equation models, accounting for within-subject correlation, were used to identify principal components that were associated with chronic ICP (comparing pre-LP images between eyes) and/or acute ICP changes (comparing pre- and post-LP images within eyes). The pp-BM width and anterior pp-BM location were calculated directly from each image and were studied in the same manner. Principal component 1 scalar variable on pre-LP images was associated with ICP (P < 0.0005). Principal component 4 magnitude changed within eyes after LP (P = 0.003). For both principal components 1 and 4, lower ICP corresponded with a more posterior position of pp-BM. Chronic ICP was associated with both pp-BM width (6.81 μm/cm H2O; P = 0.002) and more anterior location of temporal and nasal pp-BM margins (3.41, 3.49 μm/cm H2O; P < 0.0005, 0.002). This study demonstrates a quantitative association between pp-BM shape and chronic ICP level. Changes in pp-BM shape are detectable within 1 hour of lowering ICP. pp-BM shape may be a useful marker for chronic ICP level and acute ICP changes. Further study is needed to determine how pp-BM shape changes relate to clinical markers of papilledema.


Performance and Clinimetric Properties of the Timed Up From Floor Test Completed by Apparently Healthy Community-Dwelling Older Women.
Standing up from the floor is a demanding mobility activity with important implications. The purpose of this study was to describe performance and the clinimetric properties of the Timed Up From Floor (TUFF) test completed by apparently healthy community-dwelling older women. In this observational and methodological quality study, 52 community-dwelling women, 55 years and older, were examined. Convergent and discriminant validities were examined by analyzing the correlations of TUFF test times with other mobility variables and emotional status, respectively. Validity was further examined by comparing TUFF times between age groups and fall risk groups. Interrater reliability of the TUFF test was established by comparing the times obtained by 3 raters observing the same videotaped performances. Test-retest reliability was determined by having the same 3 raters observe videos of the same participants performing the TUFF test during a second session 1 week later. The grand mean (SD) TUFF time measured by all testers on the first day was 5.8 (2.9) seconds. Convergent validity was demonstrated by significant negative (P < .001) Spearman correlations between the TUFF test and the Physical Functioning Scale of the 36-Item Short Form Health Survey (SF-36) (-0.69), usual gait speed (-0.48), fast gait speed (-0.74), and the 30-second sit-to-stand test (-0.46). Discriminant validity was indicated by a low and nonsignificant correlation (0.17) between the TUFF test and the SF-36 Emotional Well-being Scale. Known-groups validity was supported by a significant difference in the TUFF test times of 2 age groups (P = .02) and 2 fall risk groups (P < .001). The TUFF test was determined to have excellent relative interrater reliability (intraclass correlation coefficient [ICC] of 0.99) and absolute reliability (minimal detectable change [MDC95%] of 0.8 seconds). Relative test-retest reliability was excellent with ICCs of 0.88 to 0.92. Corresponding MDC95% values were large (2.4-2.8 seconds and 40.7%-45.9%). The TUFF test is an informative, reliable, and valid tool suitable for documenting mobility limitations in independent community-dwelling older women. More information regarding responsiveness is required.


Long-term use of a shark breeding ground: Three decades of mating site fidelity in the nurse shark, Ginglymostoma cirratum.
Understanding shark mating dynamics and mating site use may be vital to species management. The Dry Tortugas courtship and mating ground (DTCMG) has been known as a mating site for nurse sharks, Ginglymostoma cirratum, since 1895. In a 30-yr (1992-2021) study we have documented long-term site fidelity to this area with data from 137 adult sharks (89 female, 48 male) tagged with PIT, fin, and acoustic tags. Of 118 sharks tagged from 1993 to 2014, at least 80 (68%) returned to the DTCMG in subsequent years during the June-July mating season. Known individuals returned in up to 16 different mating seasons and over periods of up 28 years, indicating that life span extends well into the forties for this species. Of all returning sharks, 59% (N = 47) have been monitored for over 10 years and 13% (N = 10) have been monitored for over 20 years. Males arrived annually in May and June and departed in July, whereas females arrived biennially or triennially in June, with a secondary peak in site use in September and August, likely associated with thermoregulation during gestation. During the mating season, males made more frequent visits of shorter duration (median = 34 visits for 1 h per visit) to the DTCMG, whereas females made fewer visits but remained on site for longer periods (median = 12.5 visits for 4.4 h per visit). Females typically mated biennially but showed a triennial cycle in 32% of cases, with many females switching cycles at least once. This pattern would reduce the potential reproductive lifetime output of a female by 11% compared to what would be projected from a strict biennial cycle. The long-term mating site fidelity of this shark population reveals the importance of identifying and protecting mating sites for this and other elasmobranch species.


The effectiveness and risks of long-term opioid therapy for chronic pain: a systematic review for a National Institutes of Health Pathways to Prevention Workshop.
Increases in prescriptions of opioid medications for chronic pain have been accompanied by increases in opioid overdoses, abuse, and other harms and uncertainty about long-term effectiveness. To evaluate evidence on the effectiveness and harms of long-term (>3 months) opioid therapy for chronic pain in adults. MEDLINE, the Cochrane Central Register of Controlled Trials, the Cochrane Database of Systematic Reviews, PsycINFO, and CINAHL (January 2008 through August 2014); relevant studies from a prior review; reference lists; and ClinicalTrials.gov. Randomized trials and observational studies that involved adults with chronic pain who were prescribed long-term opioid therapy and that evaluated opioid therapy versus placebo, no opioid, or nonopioid therapy; different opioid dosing strategies; or risk mitigation strategies. Dual extraction and quality assessment. No study of opioid therapy versus no opioid therapy evaluated long-term (>1 year) outcomes related to pain, function, quality of life, opioid abuse, or addiction. Good- and fair-quality observational studies suggest that opioid therapy for chronic pain is associated with increased risk for overdose, opioid abuse, fractures, myocardial infarction, and markers of sexual dysfunction, although there are few studies for each of these outcomes; for some harms, higher doses are associated with increased risk. Evidence on the effectiveness and harms of different opioid dosing and risk mitigation strategies is limited. Non-English-language articles were excluded, meta-analysis could not be done, and publication bias could not be assessed. No placebo-controlled trials met inclusion criteria, evidence was lacking for many comparisons and outcomes, and observational studies were limited in their ability to address potential confounding. Evidence is insufficient to determine the effectiveness of long-term opioid therapy for improving chronic pain and function. Evidence supports a dose-dependent risk for serious harms. Agency for Healthcare Research and Quality.


The incidence of childhood IDDM in New South Wales, Australia.
To determine the incidence of insulin-dependent diabetes mellitus (IDDM) in children 0-14 years of age in the state of New South Wales, Australia, which has a total population of 5.73 million. We established a prospective register, identifying 361 incident cases over a 2-year period (1990-1991) with two independent sources of case ascertainment. The primary source was the reporting of newly diagnosed patients by physicians and diabetes educators. The secondary source was a subsidized syringe scheme. Using the capture-recapture method, ascertainment was estimated to be 99.4% complete. The age-standardized incidence rate was 14.5 per 100,000 person-years (95% confidence interval: 13.0-16.0). No significant differences were found when comparing the first and second years of the register, boys and girls, geographical areas, or Aboriginal and non-Aboriginal children. There was seasonal variation in the onset (with more cases in winter), which was evident in the 10- to 14-year age-group (P = 0.01), but not in younger age-groups. A first-degree relative was already affected in 6.9% of the cases. No significant difference was noted in the age at onset when comparing cases with and without an affected first-degree relative. The incidence of childhood IDDM in New South Wales is similar to rates found in other predominantly Anglo-Saxon populations. IDDM occurs in Aboriginal children with a frequency comparable to that in the rest of the population.


Hemodynamic design requirements for in-series thoracic artificial lung attachment in a model of pulmonary hypertension.
Recent thoracic artificial lung (TAL) prototypes have impedances lower than the natural lung. With these devices, proximal pulmonary artery (PA) to distal PA TAL attachment may be possible in patients without right ventricular dysfunction. This study examined the relationship between pulmonary system impedance and cardiac output (CO) to create TAL design constraints. A circuit with adjustable resistance and compliance (C) was attached in a PA-PA fashion with the pulmonary circulation of seven sheep with chronic pulmonary hypertension. The pulmonary system zeroth harmonic impedance modulus (Z(0)) was increased by 1, 2.5, and 4 mmHg/(L/min) above baseline. At each Z(0), C was set to 0, 0.34, and 2.1 ml/mmHg. The change in pulmonary system zeroth and first harmonic impedance moduli (ΔZ(0) and ΔZ(1)), the percent change in CO (%ΔCO), and the inlet and outlet anastomoses resistances were calculated for each situation. Results indicate that ΔZ(0) (p < 0.001) but not ΔZ(1) (p = 0.5) had a significant effect on %ΔCO and that %ΔCO = -7.45*ΔZ(0) (R(2) = 0.57). Inlet and outlet anastomoses resistances averaged 0.77 ± 0.16 and 0.10 ± 0.19 mmHg/(L/min), respectively, and the relationship between %ΔCO and TAL resistance, R(T), in mmHg/(L/min) was determined to be %ΔCO = -(7.45f)×(R(T) + 0.87), in which f = the fraction of CO through the TAL. Thus, newer TAL designs can limit %ΔCO to less than 10% if f < 0.75.


Effect of transrectal ultrasonography of the prostate on serum prostate-specific antigen levels and free/total prostate-specific antigen ratio.
To study the effect of transrectal ultrasonography (TRUS) on serum total prostate-specific antigen (PSA), free PSA and free/total (f/t) PSA ratio and to determine the reliability of serum total and free PSA levels and f/t PSA ratio after such interventions, serum free and total PSA levels and f/t PSA ratios were determined before, immediately after and 24 h after TRUS in 44 patients who were referred to our department by their general practitioner for prostate screening. The duration of TRUS procedure and prostate volume and age were evaluated to determine whether the serum PSA levels following TRUS had changed. The total PSA and free PSA levels and f/t PSA ratio did not show any significant rise immediately after DRE and TRUS. The duration of the TRUS procedure did not appear to affect the PSA values. Age and prostate volume were not correlated with the PSA levels. Our findings revealed no significant changes in f/t PSA ratio and serum free and total PSA level, suggesting that serum free and total PSA levels and f/t PSA ratio may be more reliable parameters in the early period after TRUS.


BAP1 loss is unusual in well-differentiated papillary mesothelioma and may predict development of malignant mesothelioma.
Literature on BRCA1-associated protein 1 (BAP1) expression status in well-differentiated papillary mesothelioma (WDPM) is limited. In the present study, we examined the prevalence of BAP1 loss in WDPM by immunohistochemistry with clinical correlation, along with CDKN2A deletion status by fluorescence in situ hybridization (FISH). Eight patients diagnosed as having WDPM were identified from the surgical pathology file. Adenomatoid tumors (n = 8) and malignant mesothelioma (MM) (n = 39) were included for comparison. BAP1 immunohistochemistry was performed on representative block(s) from each case. CDKN2A FISH was also performed in the WDPMs and adenomatoid tumors. Clinical information was obtained from the medical records. Three of 8 WDPM patients showed synchronous or metachronous MM. All 3 cases showed BAP1 loss in both WDPM and the matched MM. Single-nucleotide polymorphism genomic microarray (n = 3) demonstrated a similar genetic profile in the WDPM and MM components, which supports their clonal relationship. The remaining 5 WDPM cases had intact BAP1 expression and had no evidence of disease on follow-up imaging studies at 1 to 71 months (median, 35 months). All 8 adenomatoid tumors had intact BAP1 expression, whereas 17 of 39 MM had BAP1 loss. CDKN2A FISH was negative for deletion in 4 WDPMs tested (including the case that developed MM) and all 8 adenomatoid tumors. In our study, WDPM did not show CDKN2A deletion in any case. BAP1 loss was also absent in all pure WDPM cases but was identified in all WDPM with synchronous or metachronous MM. Similar genetic landscape in WDPM and MM components suggested their clonal relationship.