Physiopathology of necrobiotic xanthogranuloma with monoclonal gammopathy.
Xanthomatosis associated with monoclonal gammopathy includes hyperlipidaemic xanthoma (HX), normolipidaemic xanthoma (NX) and necrobiotic xanthogranuloma (NXG). All three pathologies are characterized by skin or visceral lesions related to cholesterol accumulation, monoclonal immunoglobulin (MIg) and hypocomplementemia. The pathophysiology underlying NXG remains unknown although the involvement of MIg is suspected. To provide further insights into the pathophysiology of NXG, we evaluated the plasma lipid phenotype, mechanisms involved in cellular cholesterol accumulation and role of MIg in an analysis of blood and plasma markers of inflammation in 16 patients with xanthomatosis [NXG (n = 8) and NX (n = 8)] associated with monoclonal IgG relative to the relevant controls. The lipid profile of patients with NXG was characterized by a low HDL-C phenotype and an abnormal distribution of HDL particles. Sera from patients with NXG induced cholesterol accumulation in human macrophages. This accumulation was due in part to a significant reduction in the HDL capacity to promote cholesterol efflux from macrophages, which was not found in the case of NX. The MIg of NXG and NX patients was tested positively by ELISA to recognize a large spectrum of lipoproteins. High plasma levels of pro-inflammatory cytokines (TNFα and IL-6), soluble cytokine receptors (sIL-6R, sTNFRI and sTNFRII), adhesion molecules (VCAM-1 and ICAM-1) and chemokines (MCP-1, IL-8 and MIP-1α) were observed in both patients with NXG and NX, revealing a specific xanthoma inflammatory signature which was inversely correlated with plasma levels of anti-inflammatory HDL. However, patients with NXG were distinguished by elevated levels of IL-15 and a marked increase in the rate of intermediate CD14++CD16+ monocytes. This study revealed that NXG is characterized by impaired macrophage lipid homeostasis associated with a systemic inflammatory profile that may result from the interaction of MIg and lipoproteins.


Malignant hypertension in IgA nephropathy was not associated with background pathological phenotypes of glomerular lesions.
It was reported that IgA nephropathy (IgAN) was the major cause of secondary malignant hypertension (MHT). However, the pathogenesis of MHT secondary to IgAN (IgAN-MHT) is unknown and its association with glomerular pathological phenotypes remains inconclusive. The present study aimed to investigate whether glomerular pathological phenotypes and anti-endothelial cell antibodies (AECA) were associated with the occurrence of IgAN-MHT. The glomerular pathological phenotypes of 45 patients with IgAN-MHT were analysed using Haas' histologic grading system of IgAN. Sera were collected from 34 of the 45 patients with IgAN-MHT, 19 patients with primary MHT, 41 patients with non-MHT IgAN and 10 healthy volunteers. AECA of both IgG and IgA isotypes were detected by western blot analysis using human umbilical vein endothelial cell lysate as antigen. In the 45 patients with IgAN-MHT, 7 (15.56%), 5 (11.11%), 13 (28.89%), 9 (20%) and 11 (24.44%) patients were graded as Hass I, II, III, IV and V, respectively. Although the severity of non-ischaemic sclerosis, crescents and mesangial proliferation were significantly different between patients with different grades, the levels of blood pressure, SCr and proteinuria at presentation were comparable. Eleven and four protein bands of endothelial proteins could be blotted by AECA-IgG and AECA-IgA in sera from patients with IgAN-MHT. The prevalences of anti-121 kD AECA-IgG (15/34) and anti-92 kD AECA-IgA (10/34) in IgAN-MHT were significantly higher than that of primary MHT, non-MHT IgAN and normal controls, respectively. Patients with anti-92 kD AECA-IgA had more severe glomerular ischaemic sclerosis (6.25-92.86%, median 39.61%) than those without (0-91.67%, median 18.18%, P = 0.035). There was no significant difference in the prevalence of AECA between IgAN-MHT patients with different background glomerular lesions. Only one IgAN patient with MHT was found with both anti-121 kD AECA-IgG and anti-92 kD AECA-IgA. No particular manifestations were found. The occurrence of IgAN-MHT was not associated with the background glomerular pathological phenotypes of IgAN. AECA might play a role in the pathogenesis of IgAN-MHT.


Assessment and management of anxiety and depression in a pediatric high-risk asthma clinic.
The aims of this study were to determine the prevalence of positive mental health (MH) screens in a pediatric high-risk asthma (HRA) clinic population, and to determine the success rate of engagement in MH services before and after adding a clinical psychologist to our multidisciplinary clinic. We hypothesized that the HRA population would have a higher prevalence of anxiety/depression symptoms than that previously reported for the general pediatric asthma population. We anticipated that the presence of an embedded psychologist in HRA clinic would facilitate successful connection to MH services. Pediatric patients in the HRA clinic were prospectively screened for anxiety and depression using validated screening instruments. Positive scores were referred for MH services. Time to MH service engagement was recorded before and after the addition of a clinical psychologist. A total of 186 patients were screened; 60% had a positive MH screen. Female sex was associated with higher median scores on both screening tools and higher likelihood of engagement in MH services. After addition of a clinical psychologist, new engagement in MH services increased (20% vs. 80%, p < 0.0001), and median time to engagement decreased (14.5 vs. 0.0 months, p = 0.003). There is a high prevalence of anxiety and depression in this pediatric HRA population. Success of engagement in MH services improved after a clinical psychologist joined our multidisciplinary team, suggesting access to care as a primary barrier to engagement.


Highly sensitive and rapid detection of acetylcholine using an ITO plate modified with platinum-graphene nanoparticles.
Determining the concentrations of acetylcholine (ACh) and choline (Ch) is clinically important. ACh is a neurotransmitter that acts as a key link in the communication between neurons in the spinal cord and in nerve skeletal junctions in vertebrates, and plays an important role in transmitting signals in the brain. A bienzymatic sensor for the detection of ACh was prepared by co-immobilizing choline oxidase (ChO) and acetylcholinesterase (AChE) on graphene matrix/platinum nanoparticles, and then electrodepositing them on an ITO-coated glass plate. Graphene nanoparticles were decorated with platinum nanoparticles and were electrodeposited on a modified ITO-coated glass plate to form a modified electrode. The modified electrode was characterized by scanning electron microscopy (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) studies. The optimum response of the enzyme electrode was obtained at pH 7.0 and 35 °C. The response time of this ACh-sensing system was shown to be 4 s. The linear range of responses to ACh was 0.005-700 μM. This biosensor exhibits excellent anti-interferential abilities and good stability, retaining 50% of its original current even after 4 months. It has been applied for the detection of ACh levels in human serum samples.


Transcriptional activation of p21(WAF¹/CIP¹) is mediated by increased DNA binding activity and increased interaction between p53 and Sp1 via phosphorylation during replicative senescence of human embryonic fibroblasts.
Although p21(WAF1/CIP1) is known to be elevated during replicative senescence of human embryonic fibroblasts (HEFs), the mechanism for p21 up-regulation has not been elucidated clearly. In order to explore the mechanism, we analyzed expression of p21 mRNA and protein and luciferase activity of full-length p21 promoter. The result demonstrated that p21 up-regulation was accomplished largely at transcription level. The promoter assay using serially-deleted p21 promoter constructs revealed that p53 binding site was the most important site and Sp1 binding sites were necessary but not sufficient for transcriptional activation of p21. In addition, p53 protein was shown to interact with Sp1 protein. The interaction was increased in aged fibroblasts and was regulated by phosphorylation of p53 and Sp1. DNA binding activity of p53 was significantly elevated in aged fibroblasts but that of Sp1 was not. DNA binding activities of p53 and Sp1 were also regulated by phosphorylation. Phosphorylation of p53 at serine-15 and of Sp1 at serines appears to be involved. Taken together, the result demonstrated that p21 transcription during replicative senescence of HEFs is up-regulated by increase in DNA binding activity and interaction between p53 and Sp1 via phosphorylation.


Lattice thermal conductivity of MgO at conditions of Earth's interior.
Thermal conductivity of the Earth's lower mantle greatly impacts the mantle convection style and affects the heat conduction from the core to the mantle. Direct laboratory measurement of thermal conductivity of mantle minerals remains a technical challenge at the pressure-temperature (P-T) conditions relevant to the lower mantle, and previously estimated values are extrapolated from low P-T data based on simple empirical thermal transport models. By using a numerical technique that combines first-principles electronic structure theory and Peierls-Boltzmann transport theory, we predict the lattice thermal conductivity of MgO, previously used to estimate the thermal conductivity in the Earth, at conditions from ambient to the core-mantle boundary (CMB). We show that our first-principles technique provides a realistic model for the P-T dependence of lattice thermal conductivity of MgO at conditions from ambient to the CMB, and we propose thermal conductivity profiles of MgO in the lower mantle based on geotherm models. The calculated conductivity increases from 15 -20 W/K-m at the 670 km seismic discontinuity to 40 -50 W/K-m at the CMB. This large depth variation in calculated thermal conductivity should be included in models of mantle convection, which has been traditionally studied based on the assumption of constant conductivity.


[Automatic collection system of waste scintillator solution using micro robots].
Industrial micro-robots are now being applied to hazardous jobs or tedious jobs. These in-house micro-robots (MELFA, RM-501, Mitsubishi Electric Inc.) were used to make a collection system of waste scintillator solution on the experimental basis. Newly developed were hands, vial holders and a system soft ware, and satisfactory results have been obtained.


Coherence-a Sticky Subject.
The concept of coherence has been applied in the quantum theory of radiation and matter as well as in classical optics. We review here the meaning of the term in each case and show how these apparently different uses of the word coherence can be understood in terms of a single concept involving interference (nonzero cross terms), minimum uncertainty, and cohering (uniform variance) of the wave packet that describes the system. Self-induced transparency, superradiance, and superconductivity are discussed as dramatic examples of coherence.


Oscillatory penetration of near-fields in plasmonic excitation at metal-dielectric interfaces.
The electric field immediately below an illuminated metal-film that is perforated with a hole array on a dielectric consists of direct transmission and scattering of the incident light through the holes and evanescent near-field from plasmonic excitations. Depending on the size and shape of the hole apertures, it exhibits an oscillatory decay in the propagation direction. This unusual field penetration is explained by the interference between these contributions, and is experimentally confirmed through an aperture which is engineered with four arms stretched out from a simple circle to manipulate a specific plasmonic excitation available in the metal film. A numerical simulation quantitatively supports the experiment. This fundamental characteristic will impact plasmonics with the near-fields designed by aperture engineering for practical applications.


Yersinia pseudotuberculosis induces transcytosis of nanoparticles across human intestinal villus epithelium via invasin-dependent macropinocytosis.
Crohn's disease is characterized by a defect in intestinal barrier function, where bacteria are considered the most important inflammation-driving factor. Enteric bacteria, including E. coli and Yersinia spp, affect tight junctions in enterocytes, but little is known about bacterial effects on the transcellular pathway. Our objective was to study the short-term effects of Y. pseudotuberculosis on uptake of nanoparticles across human villus epithelium. Monolayers of human colon epithelium-derived Caco-2 cells and biopsies of normal human ileum were studied after 2 h exposure to Y. pseudotuberculosis expressing (inv+) or lacking (inv-) the bacterial adhesion molecule, invasin. Transepithelial transport of fluorescent nanoparticles (markers of transcytosis) was quantified by flow cytometry, and mechanisms explored by using inhibitors of endocytosis. Epithelial expressions of beta1-integrin and particle uptake pathways were studied by confocal microscopy. The paracellular pathway was assessed by electrical resistance (TER), mannitol flux, and expression of tight junction proteins occludin and caludin-4 by confocal microscopy. Inv+ Y. pseudotuberculosis adhered to the apical surface of epithelial cells and induced transcytosis of exogenous nanoparticles across Caco-2 monolayers (30-fold increase, P<0.01) and ileal mucosa (268+/-47% of control; P<0.01), whereas inv bacteria had no effect on transcytosis. The transcytosis was concentration-, particle size- and temperature-dependent, and possibly mediated via macropinocytosis. Y. pseudotuberculosis also induced apical expression of beta1-integrin on epithelial cells. A slight drop in TER was seen after exposure to inv+ Y. pseudotuberculosis, whereas mannitol flux and tight junction protein expression was unchanged. In summary, Y. pseudotuberculosis induced apical expression of beta1-integrin and stimulated uptake of nanoparticles via invasin-dependent transcytosis in human intestinal epithelium. Our findings suggest that bacterial factors may initiate transcytosis of luminal exogenous particles across human ileal mucosa, thus presenting a novel mechanism of intestinal barrier dysfunction.


Night and day in the VA: associations between night shift staffing, nurse workforce characteristics, and length of stay.
In hospitals, nurses provide patient care around the clock, but the impact of night staff characteristics on patient outcomes is not well understood. The aim of this study was to examine the association between night nurse staffing and workforce characteristics and the length of stay (LOS) in 138 veterans affairs (VA) hospitals using panel data from 2002 through 2006. Staffing in hours per patient day was higher during the day than at night. The day nurse workforce had more educational preparation than the night workforce. Nurses' years of experience at the unit, facility, and VA level were greater at night. In multivariable analyses controlling for confounding variables, higher night staffing and a higher skill mix were associated with reduced LOS.


Predicting glypican-3 expression in hepatocellular carcinoma: A comprehensive analysis using combined contrast-enhanced ultrasound and clinical factors.
Glypican-3 (GPC3) has emerged as a significant marker for the diagnosis and prognosis of hepatocellular carcinoma (HCC) and has garnered considerable attention as an immunotherapeutic target. In this study, we propose a combination of preoperative contrast-enhanced ultrasound (CEUS) imaging features and clinical factors to predict the positive expression of GPC3 in HCC patients. We retrospectively included 30 cases of GPC3-negative HCC and 115 cases of GPC3-positive HCC patients who underwent conventional ultrasound and CEUS evaluation. We assessed and compared the clinical characteristics, conventional ultrasound features, and CEUS features between the two groups of HCC patients. Based on the clinical and ultrasound features between the two groups, we developed a binary logistic regression model for predicting GPC3-positive HCC. A total of 145 HCC patients were included in this study. Binary logistic regression analysis showed that AFP > 20 ng/mL (OR = 4.047; 95% CI: 1.467-11.16; p = 0.007), arterial phase hyperenhancement (APHE) (OR = 12.557; 95% CI: 3.608-43.706; p < 0.001), and asynchronous perfusion (OR = 4.209; 95% CI: 1.206-14.691; p = 0.024) were predictive factors for GPC3-positive HCC. Receiver operating characteristic (ROC) analysis was conducted to predict GPC3-positive expression. The model combining the three independent predictive factors showed good predictive performance (AUC 0.817, 95% CI: 0.731-0.902, sensitivity: 91.3%, specificity: 60.0%). This combined model demonstrated excellent discriminatory ability to predict GPC3-positive HCC. Preoperative integration of CEUS features and clinical factors can non-invasively and effectively identify GPC3-positive HCC patients, providing valuable assistance in making personalized treatment decisions.


Effects of short-term JP-8 jet fuel exposure on cell-mediated immunity.
The U.S. Air Force has implemented the widespread use of JP-8 jet fuel in its operations, although a thorough understanding of its potential effects upon exposed personnel is unclear. Exposure to environmental toxicants such as JP-8 may have significant effects on host physiology. Jet fuel exposure has been shown to cause human liver dysfunction, abnormal electroencephalograms, shortened attention spans, and decreased sensorimotor speed. Previous studies have shown that short-term, low-concentration JP-8 exposure had significant effects on the immune system; e.g., decreased viable immune cell numbers, decreased immune organ weights, and loss of immune function that persisted for extended periods of time (i.e., up to 4 weeks post-exposure). In the current study, an in-depth analysis of the effects of JP-8 exposure on cellular immunity was performed. Short-term (7 days, 1 h/day), low-concentration (1000 mg/m3) exposures were conducted in mice, and T cell and natural killer (NK) cell functions were analyzed 24 h after the last exposure. The exposure regimen was found to almost completely ablate NK cell function, as well as significantly suppress the generation of lymphokine-activated killer (LAK) cell activity. Furthermore, JP-8 exposure suppressed the generation of cytotoxic T lymphocyte (CTL) cells from precursor T cells, and inhibited helper T cell activity. These findings demonstrate that JP-8 jet fuel exposure has significant detrimental effects on immune functions of exposed individuals. JP-8 jet fuel should be considered a potential and significant immunotoxicant. Chronic exposure to JP-8 may have serious implications to the long-term health of exposed individuals.


Spectrally resolved single-molecule electrometry.
Escape-time electrometry is a recently developed experimental technique that offers the ability to measure the effective electrical charge of a single biomolecule in solution with sub-elementary charge precision. The approach relies on measuring the average escape-time of a single charged macromolecule or molecular species transiently confined in an electrostatic fluidic trap. Comparing the experiments with the predictions of a mean-field model of molecular electrostatics, we have found that the measured effective charge even reports on molecular conformation, e.g., folded or disordered state, and non-uniform charge distribution in disordered proteins or polyelectrolytes. Here we demonstrate the ability to use the spectral dimension to distinguish minute differences in electrical charge between individual molecules or molecular species in a single simultaneous measurement, under identical experimental conditions. Using one spectral channel for referenced measurement, this kind of photophysical distinguishability essentially eliminates the need for accurate knowledge of key experimental parameters, otherwise obtained through intensive characterization of the experimental setup. As examples, we demonstrate the ability to detect small differences (∼5%) in the length of double-stranded DNA fragments as well as single amino acid exchange in an intrinsically disordered protein, prothymosin α.


Germline BRCA2 variants in advanced pancreatic acinar cell carcinoma: A case report and review of literature.
Pancreatic acinar cell carcinoma (PACC) is a rare tumor. Up to 45% of PACCs have alterations in the DNA damage repair pathway and 23% harbor rearrangements in the BRAF or RAF1 genes. We present a PACC case with a germline BRCA2 likely pathogenic variant (LPV) to highlight the impact of genomic testing on treatment decisions and patient outcomes. In our larger case series, we provide clinic-based information on additional 10 PACC patients treated in our center. A 70-year-old male was diagnosed with advanced PACC. At presentation, he was cachectic with severe arthralgia despite prednisolone and a skin rash that was later confirmed to be panniculitis. He was treated with modified FOLFIRINOX (mFFX) with the knowledge of the germline BRCA2 LPV. Following 11 cycles of mFFX, a computed tomography (CT) scan demonstrated significant tumor response in the pancreatic primary and hepatic metastases, totaling 70% from baseline as per Response Evaluation Criteria in Solid Tumors. Resolution of the skin panniculitis was also noted. We identified two additional PACCs with druggable targets in our case series. Our data contribute to practical evidence for the value of germline and somatic profiling in the management of rare diseases like PACC. This patient and others in our larger case series highlight the importance of genomic testing in PACC with potential utility in personalized treatment.


Identification of apoptotic tyrosine-phosphorylated proteins after etoposide or retinoic acid treatment.
A main shortcoming of using HL-60 cells as a model of granulocyte-macrophage differentiation is that some cells in the differentiating population undergo apoptosis. To address this issue, we have identified which tyrosine-phosphorylated proteins are involved in apoptosis and differentiation, respectively. HL-60 cells were induced specifically to undergo apoptosis with 68 microM etoposide, and to undergo granulocytic differentiation with 1 microM retinoic acid (RA). The corresponding two-dimensional electrophoretic maps of tyrosine-phosphorylated proteins from treated cells were compared. In the 8 h etoposide-treated HL-60 cell population, 83% of the cells were apoptotic. In the 120 h RA-treated cells, 50% of the cells were apoptotic. Eighteen cytosolic and nuclear tyrosine-phosphorylated proteins were found in both the 8 h etoposide- and the 120 h RA-treated cells, but not in the proliferating HL-60 cell population. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry analyses suggested that some of the proteins may be involved in signal transduction pathways (NFkappaB, GTP-binding protein, protein disulfide isomerase, Cyclophilin A), others in cell transcriptional and translational control (hnRNP H, hnRNP L, Hsp60, Hp1, Hcc-1, 26S proteasome beta-subunit, ATP synthase beta-chain), and a third group in cell cytoskeleton organization and receptor cycling (profilin, caveolin-1). An understanding of signal transduction in apoptosis initiation by screening for tyrosine-phosphorylated proteins associated with apoptosis may provide new targets for the treatment of leukemia.


Virtual reality efficiency as exposure therapy for alcohol use: A systematic literature review.
Virtual reality is an immersive technology that can be used as a tool in the treatment of disorders linked to substance use disorders, such as alcohol use disorder. This systematic review of the literature examines the effectiveness of virtual reality as exposure therapy for heavy social drinkers, defined as people who regularly consume alcohol in a variety of social contexts, with or without a diagnosis of alcohol use disorder. The current review includes ten studies with a total of 377 participants. Most participants were adult men (61.03%), with an age average of 44.1 years [± 7.42] and alcohol use ranging from light to heavy. Although studies show heterogeneous results, the use of virtual reality cue exposure therapies has shown greater improvement in terms of craving reduction for patients suffering from alcohol use disorder. Studies have also shown that the realism of the virtual environment can influence levels of craving and anxiety, both in heavy social drinkers. In addition, the use of virtual reality has proven to increase feeling of self-efficacy and decrease the tendency to engage in automatic drinking behaviors. However, the review also mentions the necessity of larger research to determine the efficiency of virtual reality as a therapeutic treatment for alcohol use disorder, whilst considering comorbidities and treatment background, especially for resistant patients.


Major common problems: genital prolapse.
Genital prolapse commonly results from fascial tears or muscle laxity in the pelvic floor associated with childbirth. Increasing age and raised intra-abdominal pressures often provide the additional stresses that culminate in symptomatic descent of the vaginal walls and uterus. Careful preoperative assessment of both the whole patient and the local condition is essential in the preparation for appropriate treatment. Physiotherapy plays a major role in prevention and cure when muscle laxity is the predominant causative factor. Surgical repair, despite a high recurrence rate and various complications, has become the most fashionable form of definitive treatment in all but the most trivial of cases.


DNAJ proteins and protein aggregation diseases.
Many neurodegenerative diseases are late onset diseases, associated with aggregation of proteins, implying that aged cells are more susceptible to proteotoxic stress. It is known that with aging, there is a decline in the functionality of chaperone networks and on the other hand, accumulation of damaged proteins occurs. Together, this has a cumulative effects on cellular protein homeostasis. Several studies have revealed that availability of DNAJ proteins, the co-chaperones to the Hsp70 machine, could be a rate-limiting factor in handling diseased proteins within the cell. In this review,we highlight how DNAJ proteins can affect aggregation of disease-causing proteins, if and how this depends on their function as Hsp70 co-chaperones, and how much this depends on the type of protein causing the disease. Finally, we will discuss the five known degenerative diseases that are linked to mutations in individual DNAJ members and what mechanism may underlie these DNAJ chaperonopathies.


School allergy training promotes internal policy review and enhances staff's preparedness in managing pupils with food allergy.
Recently non-statutory allergy management guidance for schools has been produced in the United Kingdom; however, there has been limited progress in implementing this. The aim of this study was to evaluate the effect of face-to-face training on self-reported school staff preparedness in managing the severely allergic child and whether it would stimulate schools' allergy policy review. A preparedness survey was conducted prior and 2 months post-intervention to assess the effect of training on self-reported preparedness and perceived confidence to manage children with food allergies. A sample of 18 primary schools that consented to participate were selected. Of the trained schools, 89% of the head teachers felt confident in dealing with an allergy emergency compared to 39% prior training (p = 0.016). Post-intervention all but one had arranged/were considering introducing allergy awareness sessions to help pupils manage their allergies (45% pre-training vs. post-training 93%, p = 0.003). Preventative measures for accidental exposure to food allergens (i.e., no food sharing policy) were adopted by all (pre-training 61% vs. post-training 100%, p = 0.03). A face-to-face school allergy training programme enhances self-reported staff preparedness and promotes internal allergy policy review in managing the needs of these children, hence addressing the current gap between recommendations and practice in schools.


Cut-offs for thyroid peroxidase and thyroglobulin antibodies in early pregnancy.
Thyroid disease in women of reproductive age is mainly of autoimmune origin, and thyroid peroxidase antibodies (TPO-Ab) as well as thyroglobulin antibodies (Tg-Ab) are key markers. Adding to this, much focus in pregnancy is on euthyroid women who are thyroid antibody positive. Evidence to substantiate the cut-offs for the definition of thyroid autoantibody positivity in early pregnant women is warranted. Stored serum samples from 14,030 Danish pregnant women were used for the measurement of TPO-Ab, Tg-Ab, TSH, and free thyroxine (ADVIA Centaur XPT, Siemens Healthineers). Among all women, a reference cohort of 10,905 individuals was identified for the establishment of antibody cut-offs. Percentile cut-offs for TPO-Ab and Tg-Ab were determined using regression on order statistics (the reference cohort). The established cut-offs were then applied (the full cohort), and frequencies of early pregnancy as well as later diagnosis of hypothyroidism were evaluated. The highest established cut-offs (95th, 97.5th, and 99th percentiles) were 59, 68, and 81 U/mL for TPO-Ab and 33, 41, and 52 U/mL for Tg-Ab. When the cut-offs were applied in the full cohort, 11.0, 10.2, and 9.7% were TPO-Ab positive, whereas 13.3, 12.3, and 11.2% were Tg-Ab positive. Antibody-positive women (TPO-Ab and/or Tg-Ab) had higher median TSH and were more likely to have hypothyroidism in early pregnancy and to be diagnosed with hypothyroidism during follow-up. This large study established and evaluated pregnancy-specific cut-offs for TPO-Ab and Tg-Ab. The findings are important regarding the classification of exposure in pregnancy and assessment of thyroid autoimmunity per se.


Perinatal alcohol use among young women living with HIV in South Africa: Context, experiences, and implications for interventions.
Perinatal alcohol use is common in South Africa, including among young women living with HIV (WLHIV), but there are few insights into the drivers of alcohol use in this population. Following the completion of a pilot trial of a peer support intervention for WLHIV aged 16-24 years in Cape Town, we purposively selected participants who had reported perinatal alcohol use at ≥1 study visits to complete a qualitative in-depth interview exploring their experiences of substance use. Of 119 women enrolled, 28 reported alcohol use, and 24 were interviewed, with ≥1/3 reporting drinking throughout their pregnancy. Women described living in a community where heavy perinatal alcohol consumption is normalised, including among their peers, leading to social pressure. Despite being aware of the risks of perinatal alcohol use, women described a disconnect between public health messaging and their experiences. Although most acknowledged the negative effects of alcohol in their lives, self-efficacy to reduce consumption was diminished by peer influences and the lack of formal employment and opportunities for recreation. These findings provide insights into the drivers of perinatal alcohol use in this setting, and suggest that without meaningful community-level changes, including employment opportunities and alternatives for socialising, interventions may have limited impact.


The yeast heat shock response is induced by conversion of cells to spheroplasts and by potent transcriptional inhibitors.
We report here that procedures commonly used to measure transcription and mRNA decay rates in Saccharomyces cerevisiae induce the heat shock response. First, conversion of cells to spheroplasts with lyticase, a prerequisite for nuclear runoff transcription, induces the expression of HSP70 and HSP90 heat shock genes. The transcript levels of the non-heat-shock gene ACT1 are slightly depressed, consistent with the general yeast stress response. Second, the DNA intercalator, 1,10-phenanthroline, widely employed as a general transcriptional inhibitor in S. cerevisiae, enhances the mRNA abundance of certain heat shock genes (HSP82, SSA1-SSA2) although not of others (HSC82, SSA4, HSP26). Third, the antibiotic thiolutin, previously demonstrated to inhibit all three yeast RNA polymerases both in vivo and in vitro, increases the RNA levels of HSP82 5- to 10-fold, those of SSA4 greater than 25-fold, and those of HSP26 greater than 50-fold under conditions in which transcription of non-heat-shock genes is blocked. By using an episomal HSP82-lacZ fusion gene, we present evidence that lyticase and thiolutin induce heat shock gene expression at the level of transcription, whereas phenanthroline acts at a subsequent step, likely through message stabilization. We conclude that, because of the exquisite sensitivity of the yeast heat shock response, procedures designed to measure the rate of gene transcription or mRNA turnover can themselves impact upon each process.


Ultrasound and PET-CT Correlation in Shoulder Pathology: A 5-Year Retrospective Analysis.
To correlate shoulder ultrasound and radiography with F-FDG PET-CT to establish FDG uptake and therefore range of metabolic activity, as defined by SUV analysis, in various symptomatic shoulder pathologies. Retrospective database query was performed for shoulder ultrasound and PET-CT scans between January 2012 and January 2017. Patients who had both studies within 1 year were included. Age- and sex-matched control patients with PET-CT scans only were also included. Retrospective image review determined shoulder pathology, and F-FDG SUVmax was measured using regions of interest placed at the glenohumeral joint, rotator cuff/bursa, and bicipital groove. Glenohumeral and acromioclavicular osteoarthrosis was assessed by radiography using the Kellgren-Lawrence classification system. Thirty-three patients had both imaging studies within 1 year. Ten patients (11 cases) were included, ranging in age from 56 to 90 years (mean, 67.9 years). Control subjects were selected among patients receiving PET-CT within 1 week of symptomatic patients. Glenohumeral osteoarthrosis was mild in 3 (27%), moderate in 2 (18%), and severe in 2 (18%). Six full-thickness rotator cuff tears (55%) were identified. SUVmax means were compared between the pathologic and control groups and were significantly higher in the former: glenohumeral joint (1.96 vs 1.32; P = 0.016), rotator cuff/bursa (2.80 vs 2.0; P = 0.005), and bicipital groove (2.19 vs 1.48; P = 0.007). The highest values were seen in full-thickness rotator cuff tear and severe biceps tenosynovitis. Increased metabolic activity about the shoulder is associated with a spectrum of rotator cuff, glenohumeral joint, and other soft tissue pathology that can be correlated with diagnostic ultrasound findings.


Anorexia nervosa in later life.
Anorexia nervosa is generally considered a disease of adolescence. However, recently there has been a growing number of case reports of anorexia nervosa occurring in later life. Physicians must be aware of the fact that anorexia nervosa can occur well into the fifth, sixth, and seventh decades of life. Three cases of probable anorexia nervosa in later life are presented. When confronted with cases of unexplained weight loss, physicians should include anorexia nervosa in their differential diagnosis and pursue the answers to questions that will help make the diagnosis.


En route to terpene natural products utilizing supramolecular cyclase mimetics.
Covering: literature up to 2018 Terpenes are a class of natural products characterized by remarkable structural diversity. Much of this diversity arises biosynthetically from a handful of linear precursors through the so-called tail-to-head terpene cyclization reaction. This reaction is one of the most complex observed in nature, and historically attempts to replicate it with non-enzymatic means have met with little success. In recent years, however, the development of manmade binding pockets that allow such reactions to take place has been reported. This Highlight provides an overview of this nascent field, and outlines the challenges that need to be overcome moving forward.


Outcome at forty-five years after open reduction and innominate osteotomy for late-presenting developmental dislocation of the hip.
A consecutive series of seventy-six patients (101 hips) underwent primary open reduction, capsulorrhaphy, and innominate osteotomy for late-presenting developmental dislocation of the hip. They were between 1.5 and five years old at the time of surgery, which was done between 1958 and 1965. The present study was designed to review the outcome of these patients into middle age. We located and reviewed the cases of sixty patients (eighty hips), which represents a 79% rate of follow-up at forty to forty-eight years postoperatively. Nineteen patients (twenty-four hips) had undergone total hip replacement, and three (three hips) had died of unrelated causes. The remaining thirty-eight patients (fifty-three hips) were assessed by the WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) and Oxford hip score questionnaires, physical examination, and a standing anteroposterior pelvic radiograph. The radiographs were analyzed to determine the minimum joint space width and the Kellgren and Lawrence score. Accepted indices of hip dysplasia were measured. With use of Kaplan-Meier survival analysis and with the end point defined as total hip replacement, the survival rates at thirty, forty, and forty-five years after the reduction were 99% (95% confidence interval, +/-2.4%), 86% (95% confidence interval, +/-6.9%), and 54% (95% confidence interval, +/-16.4%), respectively. The average Oxford hip score and WOMAC score for the surviving hips were 16.8 (range, 0 to 82) and 16.7 (range, 0 to 71), respectively. Of the fifty-one hips for which radiographs were available, thirty-eight demonstrated a minimum joint space width of >2.0 mm and thirteen demonstrated definite osteoarthritis on the basis of this criterion. Osteoarthritis, according to the system of Kellgren and Lawrence, was grade 0 or 1 in twenty-nine hips, grade 2 in seven hips, and grade 3 or 4 in fifteen hips. The average center-edge and acetabular angles were 40 degrees (range, 0 degrees to 61 degrees ) and 32 degrees (range, 20 degrees to 43 degrees ), respectively. With the numbers studied, no significant association was detected between outcome and the modifiable risk factors of body mass index or age at the time of surgery. Hips in patients with bilateral involvement were at significantly greater risk of failure (p = 0.02). This method of treatment achieves a 54% rate of survival of the hip at forty-five years. Two-thirds of the surviving hips have an excellent prognosis forty to forty-eight years after the index procedure according to the Kellgren and Lawrence score.


Recombinant human PDCD5 (rhPDCD5) protein is protective in a mouse model of multiple sclerosis.
In multiple sclerosis (MS) and its widely used animal model, experimental autoimmune encephalomyelitis (EAE), autoreactive T cells contribute importantly to central nervous system (CNS) tissue damage and disease progression. Promoting apoptosis of autoreactive T cells may help eliminate cells responsible for inflammation and may delay disease progression and decrease the frequency and severity of relapse. Programmed cell death 5 (PDCD5) is a protein known to accelerate apoptosis in response to various stimuli. However, the effects of recombinant human PDCD5 (rhPDCD5) on encephalitogenic T cell-mediated inflammation remain unknown. We examined the effects of intraperitoneal injection of rhPDCD5 (10 mg/kg) on EAE both prophylactically (started on day 0 post-EAE induction) and therapeutically (started on the onset of EAE disease at day 8), with both of the treatment paradigms being given every other day until day 25. Repeated measures two-way analysis of variance was used for statistical analysis. We showed that the anti-inflammatory effects of rhPDCD5 were due to a decrease in Th1/Th17 cell frequency, accompanied by a reduction of proinflammatory cytokines, including IFN-γ and IL-17A, and were observed in both prophylactic and therapeutic regimens of rhPDCD5 treatment in EAE mice. Moreover, rhPDCD5-induced apoptosis of myelin-reactive CD4+ T cells, along with the upregulation of Bax and downregulation of Bcl-2, and with activated caspase 3. Our data demonstrate that rhPDCD5 ameliorates the autoimmune CNS disease by inhibiting Th1/Th17 differentiation and inducing apoptosis of predominantly pathogenic T cells. This study provides a novel mechanism to explain the effects of rhPDCD5 on neural inflammation. The work represents a translational demonstration that rhPDCD5 has prophylactic and therapeutic properties in a model of multiple sclerosis.


Silymarin attenuates mycophenolate mofetil-induced duodenal disorders in rats.
The protective effect of silymarin (SMN) on mycophenolate mofetil (MMF)-induced duodenal disorders was investigated. Forty-two Wistar rats were assigned to seven groups including control and test groups. The control animals received saline and the test animals were treated with MMF (30 mg/kg, orally) and saline, MMF and SMN (25, 50, and 100 mg/kg, orally), MMF and Celecoxib (CLX, 50 mg/kg, orally), and MMF and SMN plus CLX for 14 consecutive days. The antioxidant status and myeloperoxidase activity were determined and the histopathological examinations on duodenal section also were performed. Biochemical analyses revealed that SMN and CLX individually and in combination therapy could reduce the MMF-increased nitric oxide (NO) content, myeloperoxidase (MPA) activity, and malondialdehyde (MDA) level, while the MMF-reduced level of total thiol molecules (TTM) was increased significantly (p<0.05) by given compounds. Concurrent administration of SMN and CLX resulted in a synergistic effect on the reduction of MDA level and MPO activity. SMN and CLX were able to improve the MMF-induced histopathological damages including the villus atrophy and inflammatory cells infiltration. Our data suggest that the MMF-induced duodenal disorders may attribute to the elevated NO and MDA levels and myeloperoxidase activity that resulted in pathological injuries. Moreover, the biochemical alterations and histopathological injuries due to MMF administration were reduced by SMN alone or in combination with CLX indicating its protective effect.


Cholesterol down-regulates BK channels stably expressed in HEK 293 cells.
Cholesterol is one of the major lipid components of the plasma membrane in mammalian cells and is involved in the regulation of a number of ion channels. The present study investigates how large conductance Ca(2+)-activated K(+) (BK) channels are regulated by membrane cholesterol in BK-HEK 293 cells expressing both the α-subunit hKCa1.1 and the auxiliary β1-subunit or in hKCa1.1-HEK 293 cells expressing only the α-subunit hKCa1.1 using approaches of electrophysiology, molecular biology, and immunocytochemistry. Membrane cholesterol was depleted in these cells with methyl-β-cyclodextrin (MβCD), and enriched with cholesterol-saturated MβCD (MβCD-cholesterol) or low-density lipoprotein (LDL). We found that BK current density was decreased by cholesterol enrichment in BK-HEK 293 cells, with a reduced expression of KCa1.1 protein, but not the β1-subunit protein. This effect was fully countered by the proteasome inhibitor lactacystin or the lysosome function inhibitor bafilomycin A1. Interestingly, in hKCa1.1-HEK 293 cells, the current density was not affected by cholesterol enrichment, but directly decreased by MβCD, suggesting that the down-regulation of BK channels by cholesterol depends on the auxiliary β1-subunit. The reduced KCa1.1 channel protein expression was also observed in cultured human coronary artery smooth muscle cells with cholesterol enrichment using MβCD-cholesterol or LDL. These results demonstrate the novel information that cholesterol down-regulates BK channels by reducing KCa1.1 protein expression via increasing the channel protein degradation, and the effect is dependent on the auxiliary β1-subunit.


Implementation of Whole Slide Imaging for Clinical Purposes: Issues to Consider From the Perspective of Early Adopters.
- There is growing interest in the use of digital pathology, especially whole slide imaging, for diagnostic purposes. Many issues need to be considered when incorporating this technology into a clinical laboratory. The College of American Pathologists (CAP) established a Digital Pathology Committee to support the development of CAP programs related to digital pathology. One of its many initiatives was a panel discussion entitled "Implementing Whole-Slide Imaging for Clinical Use: What to Do and What to Avoid," given for 3 years at the CAP annual meetings starting in 2014. - To review major issues to consider when implementing whole slide imaging for clinical purposes as covered during the panel discussion. - The views expressed and recommendations given are based primarily on the personal experience of the authors as early adopters of this technology. It is not intended to be an exhaustive review of digital pathology. - Implementation is best approached in phases. Early efforts are directed toward identifying initial clinical applications and assembling an implementation team. Scanner selection should be based on intended use and budget. Recognizing pathologist concerns over the use of digital pathology for diagnostic purposes, ensuring adequate training, and performing appropriate validation studies will enhance adoption. Once implemented, the transition period from glass slide to image-based diagnostics will be associated with challenges, especially those related to a hybrid glass slide-digital slide workflow. - With appropriate preparation, planning, and stepwise implementation, whole slide imaging can be used safely and reliably for frozen sections, consultation, quality assurance, and primary diagnosis.


Anxiety and anger immediately prior to myocardial infarction and long-term mortality: Characteristics of high-risk patients.
Acute high levels of anger and anxiety are associated with an elevated risk of myocardial infarction (MI) in the following two hours. MIs preceded by these acute negative emotions may also have a poor long-term prognosis, but information about high-risk patients is lacking. We examined whether young age and female sex are associated with MIs that are preceded by negative emotions and whether age and sex moderate the subsequent increased mortality risk following MI preceded by negative emotions. We conducted a secondary analysis of the Determinants of Myocardial Infarction Onset Study (N=2176, mean age=60.1±12.3years, 29.2% women). Anxiety and anger immediately prior to (0-2h) MI and the day before (24-26h) MI were assessed using a structured interview. Subsequent 10-year all-cause mortality was determined using the US National Death Index. Anxiety during the 0-2h pre-MI period was associated with younger age (OR=0.98,95% CI=0.96-0.99 per year) and female sex (OR=1.50,95% CI=1.11-2.02). Anger in the 0-2h pre-MI period was also associated with younger age (OR=0.95,95% CI=0.94-0.96) but not with sex (OR=0.93,95% CI=0.67-1.28). During follow-up, 580 (26.7%) patients died. Mortality rate was higher if MI occurred immediately after high anxiety, particularly in patients ≥65years (HR=1.80,95% CI=1.28-2.54) vs. younger patients (HR=0.87,95% CI=0.55-1.40; p-interaction=0.015). Other interactions with sex or anger were not significant. Patients with high anxiety or anger levels in the critical 2-hour period prior to MI are younger than those without such emotional precipitants. In addition, pre-MI anxiety is associated with an elevated 10-year mortality risk in patients aged ≥65years.


Validity of wrist worn accelerometers and comparability between hip and wrist placement sites in estimating physical activity behaviour in preschool children.
Wrist-worn accelerometers can increase compliance with wearing accelerometers, however, several large scale studies continue to use hip-worn accelerometers and it is unclear how comparable data is from the two sites. The study aims were: to investigate agreement between wrist- and hip- worn accelerometers and to determine the validity of Johansson et al cut-points for wrist worn accelerometers in preschool children. A sample of 32 preschool children (21 boys, 4.2 (0.5) years, BMI 16.6 (1.1)) were videoed wearing GT3X+ accelerometers on their wrist and hip while they engaged in 1 h of free-play in their nursery. Children's activity were coded using, the children's activity rating scale (CARS): with CARS, level 1 'sedentary' and levels 2-5 were classified as time spent in total physical activity (TPA). Accelerometry data were processed using Johansson et al cut-points for the wrist data and Evenson et al cut-points for the hip data, into time spent in different intensities of physical activity (PA). The mean counts per minute (cpm) from the hip and wrist were compared. There was a strong correlation between the hip and wrist cpm (r = 0.81, p < 0.01) and total count data (r = 0.83 p < 0.01), however there was a large systematic bias with wide limits of agreement. Good agreement (mean difference (LOA) 1.1 (-9.9, 12.1) was found between the CARS estimate of TPA (29.5 (10.4) mins) and the wrist estimate, using the Johansson et al cut points (28.4 (9.8) mins). There was also a reasonable agreement between the hip estimates with the Evenson et al cut-points and Johansson et al estimate (mean difference (LOA):6.3 (-8.8, 21.4) mins. In conclusion, the findings suggest that the Johansson et al (2013 Pediatr. Obes. 10 1-6) cut-points applied to wrist worn accelerometers provides a valid estimate of TPA in preschool children and have reasonable agreement with Evenson et al cut-points applied to hip accelerometers.


Polychlorinated biphenyl and polybrominated diphenyl ether profiles vary with feeding ecology and marine rearing distribution among 10 Chinook salmon (Oncorhynchus tshawytscha) stocks in the North Pacific Ocean.
Chinook salmon (Oncorhynchus tshawytscha) along the west coast of North America have experienced significant declines in abundance and body size over recent decades due to several anthropogenic stressors. Understanding the reasons underlying the relatively high levels of persistent organic pollutants (POPs) in Chinook stocks is an important need, as it informs recovery planning for this foundation species, as well for the Chinook-dependent Resident killer whales (Orcinus orca, RKW) of British Columbia (Canada) and Washington State (USA). We evaluated the influence of stock-related differences in feeding ecology, using stable isotopes, and marine rearing ground on the concentrations and patterns of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in Chinook salmon. A principal components analysis (PCA) revealed a clear divergence of PCB and PBDE congener patterns between Chinook with a nearshore rearing distribution ('shelf resident') versus a more offshore distribution. Shelf resident Chinook had 12-fold higher PCB concentrations and 46-fold higher PBDE concentrations relative to offshore stocks. Shelf resident Chinook had PCB and PBDE profiles that were heavier and dominated by more bioaccumulative congeners, respectively. The higher δ13C and δ15N in shelf resident Chinook compared to the offshore rearing stocks, and their different marine distributions explain the large divergence in contaminant levels and profiles, with shelf resident stocks being heavily influenced by land-based sources of industrial contamination. Results provide compelling new insight into the drivers of contaminant accumulation in Chinook salmon, raise important questions about the consequences for their health, and explain a major pathway to the heavily POP-contaminated Resident killer whales that consume them.


RNA-seq liver transcriptome analysis reveals an activated MHC-I pathway and an inhibited MHC-II pathway at the early stage of vaccine immunization in zebrafish.
Zebrafish (Danio rerio) is a prominent vertebrate model of human development and pathogenic disease and has recently been utilized to study teleost immune responses to infectious agents threatening the aquaculture industry. In this work, to clarify the host immune mechanisms underlying the protective effects of a putative vaccine and improve its immunogenicity in the future efforts, high-throughput RNA sequencing technology was used to investigate the immunization-related gene expression patterns of zebrafish immunized with Edwardsiella tarda live attenuated vaccine. Average reads of 18.13 million and 14.27 million were obtained from livers of zebrafish immunized with phosphate buffered saline (mock) and E. tarda vaccine (WED), respectively. The reads were annotated with the Ensembl zebrafish database before differential expressed genes sequencing (DESeq) comparative analysis, which identified 4565 significantly differentially expressed genes (2186 up-regulated and 2379 down-regulated in WED; p<0.05). Among those, functional classifications were found in the Gene Ontology database for 3891 and in the Kyoto Encyclopedia of Genes and Genomes database for 3467. Several pathways involved in acute phase response, complement activation, immune/defense response, and antigen processing and presentation were remarkably affected at the early stage of WED immunization. Further qPCR analysis confirmed that the genes encoding the factors involved in major histocompatibility complex (MHC)-I processing pathway were up-regulated, while those involved in MHC-II pathway were down-regulated. These data provided insights into the molecular mechanisms underlying zebrafish immune response to WED immunization and might aid future studies to develop a highly immunogenic vaccine against gram-negative bacteria in teleosts.


Efficacy and safety of sildenafil added to pirfenidone in patients with advanced idiopathic pulmonary fibrosis and risk of pulmonary hypertension: a double-blind, randomised, placebo-controlled, phase 2b trial.
The benefit of sildenafil in patients with advanced idiopathic pulmonary fibrosis (IPF) at risk of poor outcomes from pulmonary hypertension, whether already present or likely to develop, is uncertain. We aimed to assess the efficacy and safety of sildenafil added to pirfenidone versus placebo added to pirfenidone for 52 weeks in patients with advanced IPF and at risk of group 3 pulmonary hypertension. We did a multicentre, international, double-blind, randomised, placebo-controlled, phase 2b study at 56 university clinics, research hospitals, and tertiary sites in Canada, Europe (Belgium, Czech Republic, Germany, Greece, Hungary, Italy, the Netherlands, Spain, and Turkey), Israel, and Africa (Egypt and South Africa). Eligible patients (aged 40-80 years) had advanced IPF (carbon monoxide diffusing capacity ≤40% predicted at screening), and were at risk of group 3 pulmonary hypertension (mean pulmonary artery pressure of ≥20 mm Hg with pulmonary artery wedge pressure of ≤15 mm Hg on previous right-heart catheterisation, or intermediate or high probability of group 3 pulmonary hypertension on echocardiography as defined by the 2015 European Society of Cardiology and European Respiratory Society guidelines). Patients were randomly assigned 1:1 to oral sildenafil tablets (20 mg three times daily) or placebo, both in addition to oral pirfenidone capsules (801 mg three times daily), using a validated interactive voice-based or web-based response system with permuted block randomisation, stratified by previous right-heart catheterisation (yes or no) and forced expiratory volume in 1 s to forced vital capacity ratio (<0·8 or ≥0·8). The composite primary endpoint was disease progression, defined as either a relevant decline in 6-min walk distance, respiratory-related admission to hospital, or all-cause mortality, after 52 weeks and was assessed in the intention-to-treat population; safety was assessed in all patients who received at least one dose of the study drug. This trial is registered with ClinicalTrials.gov, NCT02951429, and is no longer recruiting. The 11-month safety follow-up is ongoing. Between Jan 13, 2017, and Aug 30, 2018, 247 patients were screened for eligibility, 177 of whom were randomly assigned to a treatment group (n=88 sildenafil; n=89 placebo) and were assessed for the primary outcome. There was no difference in the proportion of patients with disease progression over 52 weeks between the sildenafil (64 [73%] of 88 patients) and placebo groups (62 [70%] of 89 patients; between-group difference 3·06% [95% CI -11·30 to 17·97]; p=0·65). Serious treatment-emergent adverse events were reported in 54 (61%) patients in the sildenafil group and 55 (62%) patients in the placebo group. Treatment-emergent adverse events leading to mortality occurred in 22 (25%) patients in the sildenafil group and 26 (29%) in the placebo group. Addition of sildenafil to pirfenidone did not provide a treatment benefit versus pirfenidone plus placebo up to 52 weeks in patients with advanced IPF and risk of pulmonary hypertension. No new safety signals were identified with either treatment. Although the absence of a beneficial treatment effect suggests that sildenafil is not an appropriate treatment in the overall population, further research is required to establish if specific subgroups of patients with IPF might benefit from sildenafil. F Hoffmann-La Roche.


Controlling pulse stability in singularly perturbed reaction-diffusion systems.
The aim of this paper is to investigate the use of Pyragas control on the stability of stationary, localized coherent structures in a general class of two-component, singularly perturbed, reaction-diffusion systems. We use noninvasive Pyragas-like proportional feedback control to stabilize a singular pulse solution to a two-component, singularly perturbed reaction-diffusion system. We show that in a significant region of parameter space, the control can be adjusted to stabilize an otherwise unstable pulse.


Motor fluency deficits in the sequencing of actions in schizophrenia.
Many everyday activities depend on the capacity to organize and smoothly execute motor sequences. The authors tested the hypothesis that a sequencing deficit is associated with schizophrenia. They used a new method to distinguish between lower and higher order mechanisms for the impairment. The 1st task involved triggered sequences in which sensory information from 1 movement was the cue for initiation of the following movement. Results showed that the motor sequences were performed as fluently in patients as in controls. The 2nd and 3rd tasks involved sequences in which the entire movement sequence could be preplanned. Patients executed the sequences less fluently than controls but only under the condition where action sequences were required. Furthermore, the patients' fluency deficit increased with sequence complexity. The high discrimination power of Task 3 gave the authors a means to control for a potential psychometric confound involving differential discriminating power and to argue in favor of a specific higher order motor fluency deficit in patients with schizophrenia. It is suggested that basic lower order mechanisms that integrate sensory information with outgoing motor commands are preserved in schizophrenia, whereas higher order integrative mechanisms that are required for the smooth coordination of motor sequences are impaired.


Evaluation of Electron Tomography Capabilities for Shale Imaging.
Despite the advantageous resolution of electron tomography (ET), reconstruction of three-dimensional (3D) images from multiple two-dimensional (2D) projections presents several challenges, including small signal-to-noise ratios, and a limited projection range. This study evaluates the capabilities of ET for thin sections of shale, a complex nanoporous medium. A numerical phantom with 1.24 nm pixel size is constructed based on the tomographic reconstruction of a Barnett shale. A dataset of 2D projection images is numerically generated from the 3D phantom and studied over a range of conditions. First, common reconstruction techniques are used to reconstruct the shale structure. The reconstruction uncertainty is quantified by comparing overall values of storage and transport metrics, as well as the misclassification of pore voxels compared to the phantom. We then select the most robust reconstruction technique and we vary the acquisition conditions to quantify the effect of artifacts. We find a strong agreement for large pores over the different acquisition workflows, while a wider variability exists for nanometer-scale features. The limited projection range and reconstruction are identified as the main experimental bottlenecks, thereby suggesting that sample thinning, advanced holders, and advanced reconstruction algorithms offer opportunities for improvement.


The effect of context processing on different aspects of social cognition in schizophrenia.
It is well known that individuals with schizophrenia have impaired social cognition. The construct of social cognition involves several components, including perception, interpretation, and the ability to integrate context (Adolphs R. The neurobiology of social cognition. Curr Opin Neurobiol. 2001;11:231-239; Brothers L. The social brain: a project for integrating primate behavior and neurophysiology in a new domain. Concepts Neurosci. 1990;1:27-61). Importantly, a number of studies have suggested that deficits in context processing underlie cognitive dysfunction in schizophrenia (Penn DL, Corrigan PW, Bentall RP, Racenstein JM, Newman L. Social cognition in schizophrenia. Psychol Bull. 1997;121(1):114-132; Green MF, Nuechterlein KH. Should schizophrenia be treated as a neurocognitive disorder? Schizophr Bull. 1999;25:309-319). Thus, the purpose of the current study was to investigate the relationship between context processing and different aspects of social cognition in schizophrenia. Individuals with schizophrenia (n = 41) and the healthy controls (n = 32) participated in this study. The participants completed 2 sections of The Awareness of Social Inference Test: (1) social inference minimal (SI-M) and (2) social inference enriched (SI-E). They also completed face and voice emotion discrimination tasks. In addition, we used the AX-Continuous Performance Test (AX-CPT) to measure context processing and the n-back task to measure working memory more generally. AX-CPT performance in schizophrenia was positively correlated with both SI-M and SI-E performance but not with either the face or the voice discrimination. Furthermore, the correlation between AX-CPT performance and SI-M/SI-E performance was significantly stronger in individuals with schizophrenia than in controls. These results suggest that impairments in context processing are related to inferential components of social cognition in schizophrenia but not to the ability to recognition facial or vocal emotion. As such, deficits in context processing may contribute to deficits in both "hot" and "cold" aspects of cognition in schizophrenia.


Detection of piscine orthoreoviruses (PRV-1 and PRV-3) in Atlantic salmon and rainbow trout farmed in Germany.
Piscine orthoreoviruses (PRVs) are emerging pathogens causing circulatory disorders in salmonids. PRV-1 is the etiological cause of heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon (Salmo salar), characterized by epicarditis, inflammation and necrosis of the myocardium, myositis and necrosis of red skeletal muscle. In 2017, two German breeding farms for Atlantic salmon and rainbow trout (Oncorhynchus mykiss) experienced disease outbreaks with mortalities of 10% and 20% respectively. The main clinical signs were exhaustion and lethargic behaviour. During examinations, PRV-1 in salmon and PRV-3 in trout were detected for the first time in Germany. Further analyses also indicated the presence of Aeromonas salmonicida in internal tissues of both species. While PRV-1 could be putatively linked with the disease in Atlantic salmon, most of the rainbow trout suffered from an infection with A. salmonicida and not with PRV-3. Interestingly, the sequence analysis suggests that the German PRV-3 isolate is more similar to a Chilean PRV-3 isolate from Coho salmon (Oncorhynchus kisutch) than to PRV-3 from rainbow trout from Norway. This indicates a wide geographic distribution of this virus or dispersal by global trade. These findings indicate that infections with PRVs should be considered when investigating disease outbreaks in salmonids.


Field evaluation of resistance to pyriproxyfen in Bemisia tabaci (B biotype).
We determined effects of aerial sprays of the insect growth regulator pyriproxyfen on sweetpotato whitefly, Bemisia tabaci (Gennadius) (B biotype), in Arizona cotton (Gossypium spp.) fields. We measured survival for males and females from a susceptible strain and a laboratory-selected resistant strain, as well as for hybrid female progeny from crosses between the strains. Insects were exposed directly to pyriproxyfen sprays in the field or indirectly in the laboratory by rearing them on sprayed leaves collected from the field. In all tests, survival was higher for the resistant strain than the susceptible strain, but did not differ between sexes in each strain. Survival to the adult stage did not differ between eggs and nymphs directly exposed to sprays. For susceptible and hybrid individuals, survival was lower on leaves collected the day of spraying than on leaves collected 2 wk after spraying. In contrast, survival of resistant individuals did not differ based on the timing of exposure. Dominance of resistance to pyriproxyfen depended on the type of exposure. Resistance was partially or completely dominant in direct exposure bioassays and on leaves collected 2 wk after spraying (h > 0.6). Resistance was partially recessive on leaves collected the day of spraying (mean h = 0.34). Rapid evolution of resistance to pyriproxyfen could occur if individuals in field populations with traits similar to those of the laboratory-selected strain examined here were treated intensively with this insecticide.


Virtual reality intervention for older women with breast cancer.
This study examined the effects of a virtual reality distraction intervention on chemotherapy-related symptom distress levels in 16 women aged 50 and older. A cross-over design was used to answer the following research questions: (1) Is virtual reality an effective distraction intervention for reducing chemotherapy-related symptom distress levels in older women with breast cancer? (2) Does virtual reality have a lasting effect? Chemotherapy treatments are intensive and difficult to endure. One way to cope with chemotherapy-related symptom distress is through the use of distraction. For this study, a head-mounted display (Sony PC Glasstron PLM - S700) was used to display encompassing images and block competing stimuli during chemotherapy infusions. The Symptom Distress Scale (SDS), Revised Piper Fatigue Scale (PFS), and the State Anxiety Inventory (SAI) were used to measure symptom distress. For two matched chemotherapy treatments, one pre-test and two post-test measures were employed. Participants were randomly assigned to receive the VR distraction intervention during one chemotherapy treatment and received no distraction intervention (control condition) during an alternate chemotherapy treatment. Analysis using paired t-tests demonstrated a significant decrease in the SAI (p = 0.10) scores immediately following chemotherapy treatments when participants used VR. No significant changes were found in SDS or PFS values. There was a consistent trend toward improved symptoms on all measures 48 h following completion of chemotherapy. Evaluation of the intervention indicated that women thought the head mounted device was easy to use, they experienced no cybersickness, and 100% would use VR again.


Childhood tumours with a high probability of being part of a tumour predisposition syndrome; reason for referral for genetic consultation.
Recognising a tumour predisposition syndrome (TPS) in childhood cancer patients is of major clinical relevance. The presence of a TPS may be suggested by the type of tumour in the child. We present an overview of 23 childhood tumours that in themselves should be a reason to refer a child for genetic consultation. We performed a PubMed search to review the incidence of TPSs in children for 85 tumour types listed in the International Classification of Childhood Cancer third edition (ICCC-3). The results were discussed during a national consensus meeting with representative clinical geneticists from all six academic paediatric oncology centres in The Netherlands. A TPS incidence of 5% or more was considered a high probability and therefore in itself a reason for referral to a clinical geneticist. The literature search resulted in data on the incidence of a TPS in 26 tumours. For 23/26 tumour types, a TPS incidence of 5% or higher was reported. In addition, during the consensus meeting the experts agreed that children with any carcinoma should always be referred for clinical genetic consultation as well, as it may point to a TPS. We present an overview of 23 paediatric tumours with a high probability of a TPS; this will facilitate paediatric oncologists to decide which patients should be referred for genetic consultation merely based on type of tumour.


Suicide trends in South Africa, 1968--90.
A study was undertaken to investigate suicide rates and proportional mortality trends in South Africa from 1968 to 1990. Suicide rates per 100,000 population per year and suicide proportional mortality rates per year were calculated. The analyses were stratified by race, sex, and age group. Trends were interpreted using graphs depicting smoothed sex/race-specific suicide rates and proportional mortality rates over time. Regression models for suicide rates were fitted within specific age groups to test the effects of race, sex, and time. Further analysis was done within the specific age and race groups, if any interaction was found with time. There were increases in suicide rate in the young (particularly for white males) and for whites aged 65 years and above (particularly for males). The finding that suicide among the young and elderly in South Africa is increasing suggests that preventive efforts are indicated, especially for white males. Trends for young South Africans mirror those in Europe and the USA, and attention to trends in these countries may predict future trends in youth suicide in South Africa.


Serum-free medium cultivation to improve efficacy in establishment of human embryonic stem cell lines.
Serum-containing and serum-free media were used to derive human embryonic stem (HES) cells from donated oocytes and embryos. Inner cell masses (ICM) were isolated by immunosurgery. The HES cells were found to be easily obtained and expanded in a serum-free medium. The efficacy in establishing human embryonic stem cell lines improved in a serum-free medium compared with that in serum-containing media. Four HES cell lines were derived from 13 isolated ICM on mouse embryonic fibroblast feeder layers. All four cell lines possess the same characteristics and differentiating potency: normal 46, XX or 46, XY karyotype; and expressing a series of surface markers such as APase, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, but not SSEA-1. They can form embryoid bodies in suspension culture and develop teratomas comprising derivatives of three embryonic germ layers when injected into severe combined immunodeficient mice. These preliminary results suggest that serum-free cultivation may be superior to serum-containing cultivation for deriving human embryonic stem cells.


Age-related macular degeneration-associated silent polymorphisms in HtrA1 impair its ability to antagonize insulin-like growth factor 1.
Synonymous single nucleotide polymorphisms (SNPs) within a transcript's coding region produce no change in the amino acid sequence of the protein product and are therefore intuitively assumed to have a neutral effect on protein function. We report that two common variants of high-temperature requirement A1 (HTRA1) that increase the inherited risk of neovascular age-related macular degeneration (NvAMD) harbor synonymous SNPs within exon 1 of HTRA1 that convert common codons for Ala34 and Gly36 to less frequently used codons. The frequent-to-rare codon conversion reduced the mRNA translation rate and appeared to compromise HtrA1's conformation and function. The protein product generated from the SNP-containing cDNA displayed enhanced susceptibility to proteolysis and a reduced affinity for an anti-HtrA1 antibody. The NvAMD-associated synonymous polymorphisms lie within HtrA1's putative insulin-like growth factor 1 (IGF-1) binding domain. They reduced HtrA1's abilities to associate with IGF-1 and to ameliorate IGF-1-stimulated signaling events and cellular responses. These observations highlight the relevance of synonymous codon usage to protein function and implicate homeostatic protein quality control mechanisms that may go awry in NvAMD.


The Australian health care system: reform, repair or replace?
A Festshrift gives us the opportunity to look both backwards and forwards. Ken Donald's career stretches back to his intern days in 1963 and has encompassed clinical and population health, academe, clinical settings and the bureaucracy, and playing sport at state and national levels. There has been considerable change in the health care system over the period of Ken's involvement in the sector with more change to come -- where have those changes left us? This paper discusses these changes in relation to performance criteria.


Disseminated histoplasmosis in patients receiving low-dose methotrexate therapy for psoriasis.
Low-dose methotrexate sodium therapy used for nonmalignant disease has been associated with a variety of opportunistic infections with pathogens occurring in patients with defective cellular immunity. This article describes the unusual development of disseminated histoplasmosis as a probable complication of immunosuppression resulting from use of methotrexate. We report the cases of three patients in whom disseminated histoplasmosis developed while receiving low-dose methotrexate therapy for psoriasis. Disease manifestations were unusually severe in two of the three patients. All three cases were disseminated, and two cases resulted in illnesses requiring intensive medical treatment. Each patient responded appropriately to antifungal treatment, although one patient has required long-term suppressive treatment because of persistent Histoplasma antigenuria. These cases illustrate the risk for opportunistic fungal infections in patients receiving low-dose methotrexate therapy for nonmalignant diseases. Histoplasma should be added to the list of pathogens to be suspected in patients receiving such therapy.


RNA-guided assembly of Rev-RRE nuclear export complexes.
HIV replication requires nuclear export of unspliced and singly spliced viral transcripts. Although a unique RNA structure has been proposed for the Rev-response element (RRE) responsible for viral mRNA export, how it recruits multiple HIV Rev proteins to form an export complex has been unclear. We show here that initial binding of Rev to the RRE triggers RNA tertiary structural changes, enabling further Rev binding and the rapid formation of a viral export complex. Analysis of the Rev-RRE assembly pathway using SHAPE-Seq and small-angle X-ray scattering (SAXS) reveals two major steps of Rev-RRE complex formation, beginning with rapid Rev binding to a pre-organized region presenting multiple Rev binding sites. This step induces long-range remodeling of the RNA to expose a cryptic Rev binding site, enabling rapid assembly of additional Rev proteins into the RNA export complex. This kinetic pathway may help maintain the balance between viral replication and maturation.DOI: http://dx.doi.org/10.7554/eLife.03656.001.


Bioresorption of ceramic strontium-85-labeled calcium phosphate implants in dog femora. A pilot study to quantitate bioresorption of ceramic implants of hydroxyapatite and tricalcium orthophosphate in vivo.
Ceramic hydroxyapatite and tricalcium orthophosphate were radioactively labeled with their strontium-85 analogs. Porous blocks of these materials (dimensions 2.5 X 1.25 X 0.5 cm) were implanted into surgically created defects in dog femora. The fate of the implants was studied by roentgenography, radioactivity measurements, histology, and microradiography. The radioactivity over the implant site was followed for 22 weeks. Implants were retrieved after 20-25 and 50-55 weeks. Hydroxyapatite implants were not affected by biodegradation processes, while tricalcium orthophosphate implants were subject to extensive bioresorption (25%-30% in 22 weeks). Resorption debris from tricalcium orthophosphate implants was found in mononuclear phagocytes and multinuclear osteoclastlike cells. The supposition is that tricalcium orthophosphate is transformed into hydroxyapatite in a physiologic environment. Labeling of ceramic calcium phosphate implants with strontium-85 analogs offers an adequate technique to quantitate bioresorption in vivo.


An evaluation of surgery departments in community-based medical schools.
Since 1960, most newer medical schools have been community-based and have used existing community hospitals and a large number of volunteer faculty. As these schools have evolved, many have developed more traditional characteristics. To assess the positive and negative features of these new schools, chairs of departments of surgery from 21 community-based medical schools and 17 traditional schools were surveyed to acquire information on policies and practices related to faculty development, undergraduate and graduate education, hospital relationships, clinical activities, research, and relationships with community physicians. Responses were analyzed and compared. Most community-based schools have succeeded in their mission, partly at the expense of controls usually found in more traditional schools.


Novel peptide ligands with dual acting pharmacophores designed for the pathophysiology of neuropathic pain.
The conventional design of high affinity drugs targeted to a single molecule has not resulted in clinically useful therapies for pain relief. Recent reviews have suggested that newly designed analgesic drugs should incorporate multiple targets. The distributions of cholecystokinin (CCK) and CCK receptors in the central nervous system (CNS) overlap significantly with endogenous opioid systems and can be dually targeted. CCK has been shown to act as an endogenous "anti-analgesic" peptide and neuropathic pain conditions promote endogenous CCK release in CNS regions of pain modulation. Administration of CCK into nuclei of the rostral ventromedial medulla induces pronociceptive behaviors in rats. RSA 504 and RSA 601 are novel bifunctional compounds developed to target neuropathic pain by simultaneously acting as agonists at two distinct opioid receptors and antagonizing CCK receptors in the CNS. RSA 504 and RSA 601 demonstrate agonist activity in vitro and antihypersensitivity to mechanical and thermal stimuli in vivo using the spinal nerve ligation model of neuropathic pain. Intrathecal administration of RSA 504 and RSA 601 did not demonstrate antinociceptive tolerance over 7 days of administration and did not display motor impairment or sedation using a rotarod. These are the first behavioral studies that demonstrate how multi-targeted molecule design can address the pathology of neuropathic pain. These compounds with δ and μ opioid agonist activity and CCK antagonist activity within one molecule offer a novel approach with efficacy for neuropathic pain while lacking the side effects typically caused by conventional opioid therapies.


Prevalence and predictors of posttraumatic stress symptoms in utility workers deployed to the World Trade Center following the attacks of September 11, 2001.
Recent attention has begun to be focused on the effects of disaster recovery work on nonrescue workers. The goal of this study was to assess the prevalence and predictors of posttraumatic stress disorder (PTSD) and related symptoms in a population of utility workers deployed to the World Trade Center (WTC) site in the aftermath of 9/11. Utility workers deployed to the WTC site were screened at their place of employment between 10 and 34 months following the WTC attacks, utilizing both structured interviews and self-report measures. PTSD symptoms were assessed by the CAPS and the PCL; co-morbid disorders were also assessed. 2,960 individuals with complete CAPS and PCL data were included in the analyses. Eight percent of participants had symptoms consistent with full PTSD, 9.3% with subthreshold PTSD, 6% with MDD, 3.5% with GAD, and 2.5% with panic disorder. Although risk factors included psychiatric and trauma history, 51% of individuals with probable PTSD had neither; subjective perception of threat to one's life was the best predictor of probable PTSD. Extent of exposure predicted 89% of PTSD cases in those without a psychiatric or trauma history, but only 67% of cases among those with both. Nonrescue workers deployed to a disaster site are at risk for PTSD and depression. Extent of exposure affected the most vulnerable workers differently than the least vulnerable ones. These results suggest that the relationship among predictors of PTSD may be different for different vulnerability groups, and underscore the importance of screening, education, and prevention programs for disaster workers.


Plasminogen is a complement inhibitor.
Plasminogen is a 92-kDa single chain glycoprotein that circulates in plasma as a zymogen and when converted to proteolytically active plasmin dissolves preformed fibrin clots and extracellular matrix components. Here, we characterize the role of plasmin(ogen) in the complement cascade. Plasminogen binds the central complement protein C3, the C3 cleavage products C3b and C3d, and C5. Plasminogen binds to C3, C3b, C3d, and C5 via lysine residues, and the interaction is ionic strength-dependent. Plasminogen and Factor H bind C3b; however, the two proteins bind to different sites and do not compete for binding. Plasminogen affects complement action in multiple ways. Plasminogen enhanced Factor I-mediated C3b degradation in the presence of the cofactor Factor H. Plasminogen when activated to plasmin inhibited complement as demonstrated by hemolytic assays using either rabbit or sheep erythrocytes. Similarly, plasmin either in the fluid phase or attached to surfaces inhibited complement that was activated via the alternative and classical pathways and cleaved C3b to fragments of 68, 40, 30, and 17 kDa. The C3b fragments generated by plasmin differ in size from those generated by the complement protease Factor I, suggesting that plasmin-mediated C3b cleavage fragments lack effector function. Plasmin also cleaved C5 to products of 65, 50, 30, and 25 kDa. Thus, plasmin(ogen) regulates both complement and coagulation, the two central cascade systems of a vertebrate organism. This complement-inhibitory activity of plasmin provides a new explanation why pathogenic microbes utilize plasmin(ogen) for immune evasion and tissue penetration.


ABSENCE OF BATRACHOCHYTRIUM SALAMANDRIVORANS IN A GLOBAL HOTSPOT FOR SALAMANDER BIODIVERSITY.
Batrachochytrium salamandrivorans (Bsal) is an emerging fungal pathogen that affects salamander and newt populations in Asia and Europe. In the Western Hemisphere, Bsal represents a major threat to endemic amphibian populations, which have not evolved resistance to infection, and which could experience local extinction events such as those observed in European fire salamanders (Salamandra salamandra). We report findings of a survey focusing specifically on wild lungless salamanders in the southeastern US, the most biodiverse location for salamander species globally. Between May 2016 and July 2018, we conducted 25 surveys at 10 sites across three ecoregions in Tennessee, US. Using quantitative (q)PCR, we screened water samples and skin swabs from 137 salamanders in five plethodontid genera. Although single replicates of six samples amplified during qPCR cycling, no samples could be confirmed as positive for the presence of Bsal with 28S rRNA PCR and independent laboratory screening. It is probable that we found false positive results, as reported by other researchers using the same assay. We offer recommendations for future monitoring efforts.


Polymer Microring Resonators for High-Frequency Ultrasound Detection and Imaging.
Polymer microring resonators fabricated by nanoimprinting are presented as a means of ultrasound detection. Acoustic waves impinging on a ring-shaped optical resonator cause strain in the ring dimensions, modulating optical output. Basic acoustic and optical characteristics of the microring sensor are presented. Measurements at several frequencies show a high sensitivity and low noise-equivalent pressure. The angular response is determined by sensing the optoacoustic excitation of a 49 μm polyester microsphere and shows wide-angle sensitivity. A 1-D array consisting of 4 microrings is demonstrated using wavelength multiplexing for addressing each element. The high sensitivity, bandwidth, and angular response make it a potentially useful sensor platform for many applications including high-frequency ultrasonic and photoacoustic imaging.


"A deep fragrance of academia": the Australian Tobacco Research Foundation.
(1) To review the history of the tobacco industry supported Australian Tobacco Research Foundation (ATRF)(1970-1994) for evidence of the industry's use of the Foundation to further its objectives that "more research was needed" on smoking and health and to promulgate the view that nicotine was not addictive. (2) To review efforts by public health advocates to discredit the ATRF as a public relations tool used by the Australian industry. Systematic search of previously internal industry documents released through the US Master Settlement Agreement. The ATRF was headed by prestigious Australian medical scientists, with at least one considered by the industry to be "industry positive". An international ATRF symposium on nicotine was vetted by the industry and heavily attended by industry approved scientists. Following sustained criticism from the health and medical community about the industry's creation of the ATRF to further its objectives, the ATRF's scientific committee was provoked to publicly declare in 1988 that smoking was a causative agent in disease. This criticism led to growing ATRF boycotts by scientists and substandard applications, causing the industry to see the ATRF as being poor value-for-money and eventually abandoning it. The raison d'etre for the ATRF's establishment was to allow the Australian industry to point to its continuing commitment to independent medical research, with the implied corollary that tobacco control measures were premature in the face of insufficient evidence about tobacco's harms. Sustained criticism of tobacco industry funded research schemes can undermine their credibility among the scientific community.


Identification of Cassava MicroRNAs under Abiotic Stress.
The study of microRNAs (miRNAs) in plants has gained significant attention in recent years due to their regulatory role during development and in response to biotic and abiotic stresses. Although cassava (Manihot esculenta Crantz) is tolerant to drought and other adverse conditions, most cassava miRNAs have been predicted using bioinformatics alone or through sequencing of plants challenged by biotic stress. Here, we use high-throughput sequencing and different bioinformatics methods to identify potential cassava miRNAs expressed in different tissues subject to heat and drought conditions. We identified 60 miRNAs conserved in other plant species and 821 potential cassava-specific miRNAs. We also predicted 134 and 1002 potential target genes for these two sets of sequences. Using real time PCR, we verified the condition-specific expression of 5 cassava small RNAs relative to a non-stress control. We also found, using publicly available expression data, a significantly lower expression of the predicted target genes of conserved and nonconserved miRNAs under drought stress compared to other cassava genes. Gene Ontology enrichment analysis along with condition specific expression of predicted miRNA targets, allowed us to identify several interesting miRNAs which may play a role in stress-induced posttranscriptional regulation in cassava and other plants.


Development of a novel, sensitive cell-based corin assay.
Corin (atrial natriuretic peptide-converting enzyme, EC 3.4.21) is a transmembrane serine protease expressed in cardiomyocytes. Corin exerts its cardioprotective effects via the proteolytic cleavage and activation of pro-atrial natriuretic peptide (pro-ANP) to ANP. We recently described an ANP reporter cell line stably expressing the ANP receptor, a cGMP-dependent cation channel used as a real-time cGMP biosensor, and the Ca2+-sensitive photoprotein aequorin. Here, we describe the generation of a novel reporter cell line expressing the calcium biosensor GCaMP6 instead of aequorin. In contrast to the luminescence-based assay, ANP stimulation of our novel GCaMP6 reporter cell resulted in stable, long-lasting fluorescence signals. Using this novel reporter system, we were able to detect pro-ANP to ANP conversion by purified, soluble wildtype corin (solCorin), but not the active site mutant solCorin(S985A), resulting in left-shifted concentration-response curves. Furthermore, cellular pro-ANPase activity could be detected on HEK 293 cells after transient expression of wildtype corin. In contrast, corin activity was not detected after transfection with the inactive corin(S985A) variant. In supernatants from cardiomyocyte-derived HL-1 cells pro-ANP to ANP conversion could also be detected, while in HL-1 corin knockout cells no conversion was observed. These findings underline the role of corin as the pro-ANP convertase. Our novel fluorescence-based ANP reporter cell line is well-suited for the sensitive detection of corin activity, and may be used for the identification and characterization of novel corin modulators.


Impact of severe respiratory syndrome on anxiety levels of front-line health care workers.
To identify anxiety levels among front-line health care workers during the 2003 severe acute respiratory syndrome outbreak. Questionnaire survey. Regional hospital, Hong Kong. All hospital staff were given a questionnaire; administrative staff who had not had any patient contact served as controls. Levels of contact with patients who had severe acute respiratory syndrome were measured and correlated with anxiety levels as determined by the State-Trait Anxiety Inventory. Of 4252 questionnaires distributed between May and June 2003, 2040 (48.0%) were returned and 1926 (45.3%) were valid for analysis. Overall, 534 (27.7%) respondents had had contact with patients with severe acute respiratory syndrome. Anxiety scores ranged from 20 to 80, and mean (standard deviation) scores were higher among staff who had had contact with patients with severe acute respiratory syndrome than among those who had not (52.6 [10.5] versus 49.8 [10.1], respectively; P<0.01). Mean anxiety levels were higher among workmen, health care assistants, and nurses than among administrative staff controls or doctors (P<0.01). Anxiety scores were correlated with burnout scores (Pearson's correlation coefficient, 0.52-0.59) and with discomfort from wearing protective gear (0.21-0.32). Severe acute respiratory syndrome has likely stressed the public health care system. Prediction and early identification of adverse factors in a crisis situation would allow early implementation of interventions to reduce and counteract the impact of this stress.


EW-7197, a Transforming Growth Factor-Beta Type I Receptor Kinase Inhibitor, Ameliorates Acquired Lymphedema in a Mouse Tail Model.
Background: Acquired lymphedema is a common consequence of cancer surgery. Fibrosis is one of the main causes of chronic lymphedema since it hinders lymphatic regeneration and this causes a significant decrease in lymphatic flow and accumulation of excessive protein-rich fluid. The transforming growth factor-β1 (TGF-β1) signaling pathway is known in a process of wound repair and fibrosis. In our study, the purpose was to evaluate the efficacy of EW-7197, a peroral TGF-β type I receptor kinase inhibitor, in treating acquired lymphedema. Methods and Results: For lymphedema mouse tail model, we used 10- to 12-week-old female C57BL/6 mice. The skin was circumferentially excised, making a circular band followed by cauterization of lymphatic collecting vessels. Two groups were made in this study: control and treatment. The treatment group (n = 12) received a solution consisting of 0.1 mL of artificial gastric juice and 20 mg/kg EW-7197 by gavage once daily. For evaluation, tail diameter measurement, fluorescence lymphography, and immunofluorescence images were used. EW-7197 treatment ameliorates acquired lymphedema in a mouse tail model by increasing lymphangiogenesis and interstitial flow of the lymphatics by inhibition of the fibrosis. The differences in maximal tail thicknesses between the control and treatment groups were statistically significant from 2 to 4 weeks after surgery. The treatment group showed a greater number of lymphatic vessels at the surgery site than the control group. The treatment group also showed more FITC coverage area at the surgery site. Conclusion: EW-7197 treatment ameliorates acquired lymphedema in a mouse tail model by increasing lymphangiogenesis and interstitial flow.


Highly sensitive cardiac troponin T values remain constant after brief exercise- or pharmacologic-induced reversible myocardial ischemia.
Using a new precommercial high-sensitivity cardiac troponin T (hsTnT) assay, we evaluated whether hsTnT increases after reversible myocardial ischemia. In 195 patients undergoing nuclear stress testing (ST) using single-photon emission computed tomography (SPECT) for suspected ischemic heart disease, we measured hsTnT before and 18 min, 4 h, and 24 h after the stress test. Thirty patients were excluded before ST because of cardiac troponin T (cTnT) >30 ng/L (0.03 mug/L) as measured by the fourth-generation commercial test. Another 65 patients were excluded because of a combination of fixed and reversible perfusion defects (PDs) after SPECT. We studied 18 patients with reversible PDs, 41 patients with fixed PDs, and 41 patients without any PDs. Of these 100 patients, 61 received dynamic ST and 39 pharmacological ST. Median baseline hsTnT concentrations (25th, 75th percentile) were comparable in patients with reversible, fixed, and no PDs [5.57 (2.47, 12.60), 8.01 (4.55, 12.44), and 6.90 (4.63, 10.59) ng/L, respectively]. After ST, median hsTnT concentrations did not change in the reversible, fixed, or no PD groups from baseline to 18 min [-0.41 (-0.81, 0.01), 0.01 (-0.75, 0.79), and 0.36 (-0.42, 1.01) ng/L] or from baseline to 4 h [-0.56 (-1.82, 0.74), 0.24 (-0.60, 1.45), and 0.23 (-0.99, 1.15) ng/L]. Median baseline hsTnT concentrations tended to be higher in patients undergoing pharmacological vs dynamic ST; however, there were no significant increases in hsTnT concentrations after either type of ST. Elevation of cTnT is rather a consequence of irreversible myocyte death than reversible myocardial ischemia after exercise or pharmacologic myocardial ischemia.


Lymphangiogenesis Facilitates Initial Lymph Formation and Enhances the Dendritic Cell Mobilizing Chemokine CCL21 Without Affecting Migration.
Lymphatic vessels play an important role in body fluid, as well as immune system homeostasis. Although the role of malfunctioning or missing lymphatics has been studied extensively, less is known on the functional consequences of a chronically expanded lymphatic network or lymphangiogenesis. To this end, we used K14-VEGF-C (keratin-14 vascular endothelial growth factor-C) transgenic mice overexpressing the vascular endothelial growth factor C in skin and investigated the responses to inflammatory and fluid volume challenges. We also recorded interstitial fluid pressure, a major determinant of lymph flow. Transgenic mice had a strongly enhanced lymph vessel area in skin. Acute inflammation induced by lipopolysaccharide and chronic inflammation by delayed-type hypersensitivity both resulted in increased interstitial fluid pressure and reduced lymph flow, both to the same extent in wild-type and transgenic mice. Hyperplastic lymphatic vessels, however, demonstrated enhanced transport capacity after local fluid overload not induced by inflammation. In this situation, interstitial fluid pressure was increased to a similar extent in the 2 strains, thus, suggesting that the enhanced lymph vessel area facilitated initial lymph formation. The increased lymph vessel area resulted in an enhanced production of the chemoattractant CCL21 that, however, did not result in augmented dendritic cell migration after induction of local skin inflammation by fluorescein isothiocyanate. An expanded lymphatic network is capable of enhanced chemoattractant production, and lymphangiogenesis will facilitate initial lymph formation favoring increased clearance of fluid in situations of augmented fluid filtration.


The transcription factor E2A drives neural differentiation in pluripotent cells.
The intrinsic mechanisms that link extracellular signalling to the onset of neural differentiation are not well understood. In pluripotent mouse cells, BMP blocks entry into the neural lineage via transcriptional upregulation of inhibitor of differentiation (Id) factors. We have previously identified the major binding partner of Id proteins in pluripotent cells as the basic helix-loop-helix (bHLH) transcription factor (TF) E2A. Id1 can prevent E2A from forming heterodimers with bHLH TFs or from forming homodimers. Here, we show that overexpression of a forced E2A homodimer is sufficient to drive robust neural commitment in pluripotent cells, even under non-permissive conditions. Conversely, we find that E2A null cells display a defect in their neural differentiation capacity. E2A acts as an upstream activator of neural lineage genes, including Sox1 and Foxd4, and as a repressor of Nodal signalling. Our results suggest a crucial role for E2A in establishing neural lineage commitment in pluripotent cells.