A double-blind randomized placebo controlled study assessing safety, tolerability and efficacy of palmitoylethanolamide for symptoms of knee osteoarthritis.
The aim of the study was to assess the safety, tolerability and efficacy of palmitoylethanolamide (PEA) when dosed at 300 mg and 600 mg per day on symptoms of knee osteoarthritis. This was a single site, comparative, double-blind placebo controlled study in adults with mild to moderate knee osteoarthritis with 111 participants randomized to receive 300 mg PEA, 600 mg PEA or placebo each day, in divided doses b.i.d, for 8 weeks. The primary outcome was the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). The secondary outcomes were the Numerical Rating Scales (NRS) for pain, the Depression Anxiety Stress Scale (DASS), the Perceived Stress Scale (PSS), the Pittsburg Sleep Quality Index (PSQI), the Short Form Health Survey (SF-36), the use of rescue pain medication and clinical safety assessment. There was a significant reduction in the total WOMAC score in the 300 mg PEA (p = 0.0372) and the 600 mg PEA (p = 0.0012) groups, the WOMAC pain score (300 mg PEA, p = 0.0074; 600 mg PEA, p = < 0.001), the WOMAC stiffness score (PEA 300 mg, p < 0.0490; 600 mg PEA, p = 0.001) and in the WOMAC function score in the 600 mg PEA group (p = 0.033) compared to placebo. The NRS pain evaluations for "worst pain" and "least pain" were significantly reduced in the 300 mg PEA group (p < 0.001, p = 0.005) and the 600 mg PEA group (p < 0.001, p < 0.001) compared to placebo. There was a significant reduction in anxiety (DASS) in both active treatment groups (300 mg PEA, p = 0.042; 600 mg PEA group (p = 0.043) compared to placebo. There were no changes in the clinical markers and the product was well tolerated. The study demonstrated that palmitoylethanolamide may be a novel treatment for attenuating pain and reducing other associated symptoms of knee osteoarthritis. Further studies on the pharmacological basis of this anti-inflammatory effect are now required.


Daily Consumption of Synbiotic Yogurt Decreases Liver Steatosis in Patients with Nonalcoholic Fatty Liver Disease: A Randomized Controlled Clinical Trial.
Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in developed and developing countries. The use of synbiotics has been proposed as a probable management strategy for patients with NAFLD. We investigated the effects of synbiotic yogurt on hepatic steatosis and liver enzymes as primary outcomes and on oxidative stress markers, adipokine concentration, and gut peptide concentration as secondary outcomes in patients with NAFLD. In this 24-wk, open-label, randomized controlled clinical trial, 102 patients [50 men and 52 women; mean age: 40 y; body mass index (in kg/m2) (mean ± SD): 31.2 ± 4.9] were randomly assigned to 3 groups, including 2 intervention groups and 1 control group. The intervention groups consumed 300 g synbiotic yogurt containing 108 colony-forming units Bifidobacterium animalis/mL and 1.5 g inulin or conventional yogurt daily and were advised to follow a healthy lifestyle (i.e., diet and exercise). The control group was advised to follow a healthy lifestyle alone. We evaluated differences between groups in liver function measures by using repeated-measures ANOVA, ANCOVA, and logistic regression. At the end of the study, the grades of NAFLD, as determined by ultrasonography, showed a significant decrease in the synbiotic group compared with the conventional and control groups (P < 0.001). The following significant mean ± SD decreases were seen in the synbiotic, conventional, and control groups, respectively: serum concentration of alanine aminotransferase (-14.5 ± 15.6 compared with 4.6 ± 15.4 and 3.1 ± 14.4 IU/L; P = 0.008), aspartate aminotransferase (-7.5 ± 6.1 compared with 3.0 ± 8.2 and 3.1 ± 5.7 IU/L; P < 0.001), alkaline phosphatase (-26.2 ± 16.8 compared with 3.4 ± 30.1 and 1.5 ± 31.9 IU/L; P = 0.024), and γ-glutamyltransferase (-6.0 ± 6.0 compared with 1.0 ± 6.4 and 7.6 ± 11.4 IU/L; P < 0.001). Synbiotic yogurt consumption improved hepatic steatosis and liver enzyme concentrations in patients with NAFLD. This trial was registered at the Iranian Registry of Clinical Trials website (www.irct.ir) as IRCT2017020932417N2.


Significant linkage to spondyloarthropathy on 9q31-34.
Spondyloarthropathy (SpA) is a frequent rheumatologic disorder with a prevalence of 0.3% in Caucasian populations from western Europe. It commonly presents as chronic axial and/or peripheral arthritis with potential disabling outcome. SpA is also variably associated with extra-articular manifestations. The pathogenesis of SpA is considered as complex, with a strong genetic component. Human leukocyte antigen B27 has been identified as a predisposing factor for SpA, but family and twin studies suggest that additional genetic risk factors exist outside the major histocompatibility complex (MHC). To map SpA susceptibility loci, 120 multiplex SpA families were included in a genome-wide scan. Linkage analyses performed on the first 65 families allowed us to identify four candidate non-MHC regions on chromosomes 5q, 9q, 13q and 17q, which were further explored in the remaining 55 multiplex families (extension study). Non-parametric multipoint linkage analyses of the whole data set yielded evidence of significant linkage to 9q31-34, in the vicinity of marker D9S1776 (NPL=4.87, LOD=5.15, P=0.00002). This result provides evidence for the presence of a non-MHC susceptibility locus for SpA mapping to 9q31-34.


The paracrine effects of fibroblasts on Doxorubicin-treated breast cancer cells.
Breast cancer is frequently diagnosed in women and poses a major health problem throughout the world. Currently, the unresponsiveness of cancer cells to chemotherapeutics is a major concern. During chemotherapeutic treatment with Doxorubicin, neighbouring cells in the tumor microenvironment are also damaged. Depending on the concentration of Doxorubicin, apoptotic or senescent fibroblasts in the tumor microenvironment can then secrete a variety of bioactive molecules which promote tumor growth, metastasis and drug resistance. Mouse embryonic fibroblasts (MEFs) were treated with Doxorubicin to induce apoptosis and senescence respectively. Conditioned media was collected from the MEFs and was used to assess the paracrine effects between fibroblasts and E0771 murine breast cancer cells. Senescent fibroblasts significantly increased cell viability in E0771 cells following Doxorubicin treatment by activating Akt and ERK. Autophagy contributed to cancer cell death and not to treatment resistance in breast cancer cells. Our results highlight the complexity of the tumor microenvironment where chemotherapeutic agents such as Doxorubicin can induce significant changes fibroblasts which can affect tumor growth via the secretion of paracrine factors. Here we have demonstrated that those secreted paracrine factors enhance breast cancer growth and induce therapeutic resistance through the evasion of apoptotic cell death.


Development and validation of a ReaxFF reactive force field for Cu cation/water interactions and copper metal/metal oxide/metal hydroxide condensed phases.
To enable large-scale reactive dynamic simulations of copper oxide/water and copper ion/water interactions we have extended the ReaxFF reactive force field framework to Cu/O/H interactions. To this end, we employed a multistage force field development strategy, where the initial training set (containing metal/metal oxide/metal hydroxide condensed phase data and [Cu(H(2)O)(n)](2+) cluster structures and energies) is augmented by single-point quantum mechanices (QM) energies from [Cu(H(2)O)(n)](2+) clusters abstracted from a ReaxFF molecular dynamics simulation. This provides a convenient strategy to both enrich the training set and to validate the final force field. To further validate the force field description we performed molecular dynamics simulations on Cu(2+)/water systems. We found good agreement between our results and earlier experimental and QM-based molecular dynamics work for the average Cu/water coordination, Jahn-Teller distortion, and inversion in [Cu(H(2)O)(6)](2+) clusters and first- and second-shell O-Cu-O angular distributions, indicating that this force field gives a satisfactory description of the Cu-cation/water interactions. We believe that this force field provides a computationally convenient method for studying the solution and surface chemistry of metal cations and metal oxides and, as such, has applications for studying protein/metal cation complexes, pH-dependent crystal growth/dissolution, and surface catalysis.


[Evaluation of the levels of radiofrequency electromagnetic fields in the territory of the city of Bari in outside and inside environments].
In this study we measured the levels of the high frequency field in the proximity of non-ionizing radiation sources (wireless transmitting stations for mobile telephones and radio and television transmitters) in nine districts of the city of Bari. The measurements were taken both inside and outside closed environments. For the indoor measurements we took into account electromagnetic field generating equipment (VDT, electric domestic appliances, mobile telephones) in working and non-working order and with the windows open and shut respectively. We carried out these measurements according to the methods laid down in the Italian regulation CEI ENV 50166-2 of May 1995, as shown in the enclosure to the Ministerial Decree of 10.9.98 n.381. The electromagnetic field levels near wireless transmitting stations for mobile telephones are certainly modest when we consider that they never exceeded the limits established by the aforesaid Ministerial Decree. On the contrary radio and television equipment creates a much greater source of exposure. The electromagnetic field levels are certainly superior to those of the wireless transmitting stations although they never exceed, except in one isolated case, the values established by the Ministerial Decree 381/98.


Task-Evoked Dynamic Network Analysis Through Hidden Markov Modeling.
Complex thought and behavior arise through dynamic recruitment of large-scale brain networks. The signatures of this process may be observable in electrophysiological data; yet robust modeling of rapidly changing functional network structure on rapid cognitive timescales remains a considerable challenge. Here, we present one potential solution using Hidden Markov Models (HMMs), which are able to identify brain states characterized by engaging distinct functional networks that reoccur over time. We show how the HMM can be inferred on continuous, parcellated source-space Magnetoencephalography (MEG) task data in an unsupervised manner, without any knowledge of the task timings. We apply this to a freely available MEG dataset in which participants completed a face perception task, and reveal task-dependent HMM states that represent whole-brain dynamic networks transiently bursting at millisecond time scales as cognition unfolds. The analysis pipeline demonstrates a general way in which the HMM can be used to do a statistically valid whole-brain, group-level task analysis on MEG task data, which could be readily adapted to a wide range of task-based studies.


A cluster of olfactory receptor genes linked to frugivory in bats.
Diversity of the mammalian olfactory receptor (OR) repertoire has been globally reshaped by niche specialization. However, little is known about the variability of the OR repertoire at a shallower evolutionary timeframe. The vast bat radiation exhibits an extraordinary variety of trophic and sensory specializations. Unlike other mammals, bats possess a unique and diverse OR gene repertoire. We elucidated whether the evolution of the OR gene repertoire can be linked to ecological niche specializations, such as sensory modalities and diet. The OR gene repertoires of 27 bat species spanning the chiropteran radiation were amplified and sequenced. For each species, intact and nonfunctional genes were assessed, and the OR gene abundances in each gene family were analyzed and compared. We identified a unique OR pattern linked to the frugivorous diet of New World fruit-eating bats and a similar convergent pattern in the Old World fruit-eating bats. Our results show a strong association between niche specialization and OR repertoire diversity even at a shallow evolutionary timeframe.


Probability estimation of a Carrington-like geomagnetic storm.
Intense geomagnetic storms can cause severe damage to electrical systems and communications. This work proposes a counting process with Weibull inter-occurrence times in order to estimate the probability of extreme geomagnetic events. It is found that the scale parameter of the inter-occurrence time distribution grows exponentially with the absolute value of the intensity threshold defining the storm, whereas the shape parameter keeps rather constant. The model is able to forecast the probability of occurrence of an event for a given intensity threshold; in particular, the probability of occurrence on the next decade of an extreme event of a magnitude comparable or larger than the well-known Carrington event of 1859 is explored, and estimated to be between 0.46% and 1.88% (with a 95% confidence), a much lower value than those reported in the existing literature.


Low Level Laser Therapy Reduces the Development of Lung Inflammation Induced by Formaldehyde Exposure.
Lung diseases constitute an important public health problem and its growing level of concern has led to efforts for the development of new therapies, particularly for the control of lung inflammation. Low Level Laser Therapy (LLLT) has been highlighted as a non-invasive therapy with few side effects, but its mechanisms need to be better understood and explored. Considering that pollution causes several harmful effects on human health, including lung inflammation, in this study, we have used formaldehyde (FA), an environmental and occupational pollutant, for the induction of neutrophilic lung inflammation. Our objective was to investigate the local and systemic effects of LLLT after FA exposure. Male Wistar rats were exposed to FA (1%) or vehicle (distillated water) during 3 consecutive days and treated or not with LLLT (1 and 5 hours after each FA exposure). Non-manipulated rats were used as control. 24 h after the last FA exposure, we analyzed the local and systemic effects of LLLT. The treatment with LLLT reduced the development of neutrophilic lung inflammation induced by FA, as observed by the reduced number of leukocytes, mast cells degranulated, and a decreased myeloperoxidase activity in the lung. Moreover, LLLT also reduced the microvascular lung permeability in the parenchyma and the intrapulmonary bronchi. Alterations on the profile of inflammatory cytokines were evidenced by the reduced levels of IL-6 and TNF-α and the elevated levels of IL-10 in the lung. Together, our results showed that LLLT abolishes FA-induced neutrophilic lung inflammation by a reduction of the inflammatory cytokines and mast cell degranulation. This study may provide important information about the mechanisms of LLLT in lung inflammation induced by a pollutant.


The gut, its microbiome, and the brain: connections and communications.
Modern research on gastrointestinal behavior has revealed it to be a highly complex bidirectional process in which the gut sends signals to the brain, via spinal and vagal visceral afferent pathways, and receives sympathetic and parasympathetic inputs. Concomitantly, the enteric nervous system within the bowel, which contains intrinsic primary afferent neurons, interneurons, and motor neurons, also senses the enteric environment and controls the detailed patterns of intestinal motility and secretion. The vast microbiome that is resident within the enteric lumen is yet another contributor, not only to gut behavior, but to the bidirectional signaling process, so that the existence of a microbiota-gut-brain "connectome" has become apparent. The interaction between the microbiota, the bowel, and the brain now appears to be neither a top-down nor a bottom-up process. Instead, it is an ongoing, tripartite conversation, the outline of which is beginning to emerge and is the subject of this Review. We emphasize aspects of the exponentially increasing knowledge of the microbiota-gut-brain "connectome" and focus attention on the roles that serotonin, Toll-like receptors, and macrophages play in signaling as exemplars of potentially generalizable mechanisms.


Background potassium channel block and TRPV1 activation contribute to proton depolarization of sensory neurons from humans with neuropathic pain.
Protons cause a sustained depolarization of human dorsal root ganglion (DRG) neurons [Baumann et al. (1996) Pain, 65, 31-38]. In the present study we sought to determine which ion channels are expressed in human DRG neurons that could mediate the sustained responses observed in the patch-clamp recordings. RT-PCR of material from the DRG tissue revealed the presence of mRNAs for a nonselective cation channel that is activated by protons (TRPV1) and background potassium channels that are blocked by protons (TASK-1, TASK-3 and Kir2.3). Highly acidic solution (pH 5.4) applied to cultured DRG neurons evoked prolonged currents that were associated with a net increase in membrane conductance. Consistent with the involvement of TRPV1, these proton-evoked currents were blocked by capsazepine and were only found in neurons that responded to capsaicin with an increase in membrane conductance. Less acidic extracellular solution (pH 6.0) evoked such currents only rarely, but was able to strongly enhance the currents evoked by capsaicin. Capsazepine (1 microm) blocked the currents evoked by capsaicin at pH 7.35, as well as the potentiated responses to capsaicin at pH 6.0. In neurons that were not excited by capsaicin, moderate extracellular acidification (pH 6.0) caused a sustained decrease in resting membrane conductance. The decrease in membrane conductance by protons was associated with inhibition of background potassium channels. This excitatory effect of protons was not blocked by capsazepine. We conclude that in most neurons the sustained depolarization in response to moderately acidic solutions is the result of blocked background potassium channels. In a subset of neurons, TRPV1 also contributes.


Magnetoelectric coupling in nanoscale 0-1 connectivity.
The strain-mediated magnetoelectric (ME) coupling between piezoelectric (PE) and magnetostrictive (MS) components are established in various connectivities. Discovering new ME connectivity and elucidating the key factors governing the performance of ME composite are of critical importance to find advanced materials for modern electronics. Reported here is a novel ME coupling in 0-1 connectivity. The unique self-assembling ability of 1-dimension crystalline nanocellulose (CNC) nanowhiskers enables the establishment of ME coupling with 0-dimension cobalt ferrite (CFO) nanoparticles without the use of binder. The developed CFO/CNC 0-1 ME composites display a significant ME voltage coefficient (αME) as high as 0.135 mV cm-1 Oe-1. The CFO nanoparticles are also modified with a cationic surfactant, cetyltrimethylammonium bromide (CTAB), to reduce their dispersion ability. A ME response related to the rearrangement of aggregated MS nanoparticles is observed in the CTAB-CFO/CNC composites, which differs from the typical magnetostriction induced ME effect in nanoparticulate ME composites.


A neonate with tracheobronchial cobblestone mucosa presenting with acute severe hypercapnia.
Liu Y, Ly Li, Hu YL. A neonate with tracheobronchial cobblestone mucosa presenting with acute severe hypercapnia. Turk J Pediatr 2019; 61: 971-974. Here, we describe the case of a neonate suffering from respiratory distress complicated by pulmonary hemorrhage shortly after birth. The neonate experienced repeated dyspnea and acute severe hypercapnia (160-100 mmHg) that began on the 7th day after birth during invasive mechanical ventilation. Rescue high frequency oscillatory ventilation played a key role in maintaining gas exchange during these episodes. Subsequent bronchoscopy revealed tracheobronchial edematous mucosa with a cobblestone pattern, which we believe is the first report of this condition in neonates. Protrusion of the diffuse edematous mucosa led to airway stenosis and respiratory distress. Lesions of the tracheal mucosa should therefore be considered as one potential cause of neonatal respiratory distress.


Genetic ancestries in northwest Cambodia.
A survey of the genetic ancestry of 125 Cambodian children resident in Siem Reap province was undertaken, based on eight Y-chromosome binary polymorphisms and sequencing of the mtDNA HV1 region. The data indicated a largely East Asian paternal ancestry and a local Southeast Asian maternal ancestry. The presence of Y-chromosomes P* and R1al* was suggestive of a small but significant Indo-European male ancestral component, which probably reflects the history of Indian, and later European, influences on Cambodia.


Genetic variability in populations of the southern chinch bug, Blissus insularis, assessed using AFLP analysis.
Southern chinch bug, Blissus insularis Barber (Heteroptera: Blissidae), is the most destructive insect pest of St. Augustine grass, Stenotaphrum secundatum Waltz (Kuntze), in the southern United States. The present study is focused on assessing genetic variability in five populations of B. insularis collected from Texas and Florida where St. Augustine grass is widely grown. The amplified fragment length polymorphism technique was used to DNA fingerprint individuals from each population (a total of 46 individuals) using five primer combinations (EcoRI/MSeI). Analysis of molecular variance results show no evidence to support significant genetic variability among Texas and Florida populations of B. insularis. Nearly all genetic variation was found to reside within populations (95%), with only approximately 3% residing among populations between regions. Low G(ST) values obtained from POPGENE and low F(ST) values obtained from the analysis of molecular variance both support the conclusion for high levels of gene flow resulting from interbreeding and/or migratory events among the populations. A Mantel test of Euclidean squared distances showed no correlation between the genetic distance and geographic distance matrices of tested populations of B. insularis. The results of the present study suggests that gene flow is occurring among populations of B. insularis and, therefore, breeders need to be aware that this resistance will most likely not remain localized, and it has the potential to spread as a result of migratory events.


Intake of vitamin C, vitamin E, selenium, zinc and polyunsaturated fatty acids and upper respiratory tract infection-a prospective cohort study.
Antioxidants and polyunsaturated fatty acids (PUFAs) have a role in the human immune defense and may affect the susceptibility to upper respiratory tract infection (URTI). To examine dietary intake of vitamin C, vitamin E, selenium, zinc and PUFAs in relation to URTI incidence in a prospective cohort study. A total of 1533 Swedish women and men aged 25-64 years were followed for nine months during 2011-2012. Information on dietary intake was assessed through a web-based food frequency questionnaire, and events of URTI were self-reported prospectively as they occurred. Cox proportional hazards regression was applied to obtain incidence rate ratios with 95% confidence intervals. The mean number of URTI events was 0.9 among all participants, 1.0 among women and 0.7 among men. In women, the incidence rate ratios (95% confidence interval) for high compared with low intake were 0.69 (0.55-0.88) for vitamin C, 0.77 (0.62-0.96) for vitamin E, 0.57 (0.39-0.83) for docosahexaenoic acid (DHA) and 0.80 (0.65-0.99) for arachidonic acid (AA). No association was found for selenium or zinc among women. In men, an increased URTI incidence was seen with medium vitamin E intake (1.42 (1.09-1.85)) and high zinc intake (1.50 (1.04-2.16)). No association was found for vitamin C, selenium or PUFAs among men. We found an inverse association of URTI incidence among women for vitamin C, vitamin E, DHA and AA intake and a positive association among men for vitamin E and zinc intake. The observed gender differences warrant further investigation.


Epistaxis: a review of hospitalized patients.
Epistaxis is a common disease. That is usually self-limited and controlled by extemal compression. However, posterior epistaxis is occasionally complicated and needs hospitalization. The aim of this study is to analyze the etiology of epistaxis that requires hospitalization, and present the results of the treatments. From June 1997 to May 1999, 44 patients admitted under diagnosis of epistaxis were retrospectively investigated. Among these patients, 7 were admitted twice for epistaxis. The demographics, administrative history, post-history and associated diseases, treatment and complications were analyzed. Noninterventional treatment comprised of nasal packing and local electrocauterization; interventional treatment included surgery and embolization. The mean age of the patients enrolled was 53.4 years. Men outnumbered women in a rate of 3:1. The mean length for hospital stays was 8.1 days for noninterventionally-treated patients, 11.8 days for interventionally-treated patients. Emergency room visit prior to admission was noted in 68.6% of the patients. Hypertension, nasopharyngeal carcinoma (NPC) after radiotherapy, a history of nasal operations and smoking were four major associated diseases. The failure rate to control the epistaxis was 26.7% for noninterventional treatments and 16.7% for interventional treatments. The complication rate was 2% for the entire study, and the mortality rate was 7.8%. All the mortal patients were post-irradiation NPC patients with internal carotid artery bleeding. Most epistaxis patients can be managed in a noninterventional manner. Interventional treatment is only warranted in those whose epistaxis persists after adequate noninterventional treatment. For internal carotid artery epistaxis in NPC patients, embolization should be performed as soon as possible.


Humanoid Cognitive Robots That Learn by Imitating: Implications for Consciousness Studies.
While the concept of a conscious machine is intriguing, producing such a machine remains controversial and challenging. Here, we describe how our work on creating a humanoid cognitive robot that learns to perform tasks via imitation learning relates to this issue. Our discussion is divided into three parts. First, we summarize our previous framework for advancing the understanding of the nature of phenomenal consciousness. This framework is based on identifying computational correlates of consciousness. Second, we describe a cognitive robotic system that we recently developed that learns to perform tasks by imitating human-provided demonstrations. This humanoid robot uses cause-effect reasoning to infer a demonstrator's intentions in performing a task, rather than just imitating the observed actions verbatim. In particular, its cognitive components center on top-down control of a working memory that retains the explanatory interpretations that the robot constructs during learning. Finally, we describe our ongoing work that is focused on converting our robot's imitation learning cognitive system into purely neurocomputational form, including both its low-level cognitive neuromotor components, its use of working memory, and its causal reasoning mechanisms. Based on our initial results, we argue that the top-down cognitive control of working memory, and in particular its gating mechanisms, is an important potential computational correlate of consciousness in humanoid robots. We conclude that developing high-level neurocognitive control systems for cognitive robots and using them to search for computational correlates of consciousness provides an important approach to advancing our understanding of consciousness, and that it provides a credible and achievable route to ultimately developing a phenomenally conscious machine.


Parental occupational exposures and autism spectrum disorder.
Both self-report and industrial hygienist (IH) assessed parental occupational information were used in this pilot study in which 174 families (93 children with ASD and 81 unaffected children) enrolled in the Childhood Autism Risks from Genetics and Environment study participated. IH results indicated exposures to lacquer, varnish, and xylene occurred more often in the parents of children with ASD compared to the parents of unaffected children. Parents of children with ASD were more likely to report exposures to asphalt and solvents compared to parents of unaffected children. This study was limited by the small sample size, but results suggest that workplace exposures to some chemicals may be important in the etiology of ASD and deserve further investigation.


Phagocytosis of Escherichia coli biofilm cells with different aspect ratios: a role of substratum material stiffness.
Bacterial biofilms play an important role in chronic infections due to high-level tolerance to antibiotics. Thus, it is important to eradicate bacterial cells that are attached to implanted medical devices of different materials. Phagocytosis is a key process of the innate immunity to eliminate invading pathogens. Previous research demonstrated that the efficiency of phagocytosis is affected by the aspect ratio of polymer beads. Recently, we reported that the stiffness of polydimethylsiloxane (PDMS) influences Escherichia coli biofilm formation and the biofilm cells on stiff (5:1) PDMS are 46.2% shorter than those on soft (40:1) PDMS. Based on these findings, we hypothesized that E. coli cells attached on stiff PDMS can be more effectively removed via phagocytosis. This hypothesis was tested in the present study using viability assays, flow cytometry, and cell tracking. The results revealed that shorter E. coli cells detached from stiff PDMS were easier to be phagocytized than the longer cells from soft PDMS surfaces. Furthermore, macrophage cells were found to be more motile on stiff PDMS surfaces and more effective at phagocytosis of E. coli cells attached on these surfaces. These results may help the design of better biomaterials to reduce fouling and associated infections.


Single-dose pharmacokinetics of Nestorone, a potential female-contraceptive.
A synthetic progestin Nestorone is being developed for female-contraception. This study was conducted to determine the distribution, metabolism, and excretion of tritium-labeled Nestorone ((3)H Nestorone) in adult female rats. Rats were injected subcutaneously (S.C.) with a single dose of 400 microCi (3)H Nestorone/kg BW. Its distribution and concentrations in blood, plasma and other tissues were determined at defined times. The excreta were examined for elimination of (3)H Nestorone. Radioactivity in all samples was analyzed by liquid scintillation counter. Metabolite profiling was performed by HPLC and LC/MS analysis of the plasma, urine, and feces samples. Following subcutaneous injection of (3)H Nestorone, the mean peak concentrations of radioactivity (C(max)) in the blood and plasma were 58.1 and 95.5 ng equiv. (3)H Nestorone/g, respectively, at 2-h postdose (T(max)). Thereafter, the concentration of drug steadily declined through 96-h postdose with a terminal elimination half-life (t(1/2)) of 15.6 h. (3)H Nestorone-derived radioactivity was widely distributed in most tissues by 0.5 h and attained a mean maximal concentration by 2-h postdose. Approximately, 81.4% and 7.62% of the administered dose was excreted via feces and urine, respectively. In vivo metabolism of (3)H Nestorone resulted into a total of 19 metabolites. Among them, two metabolites viz., 17alpha-deacetyl-Nestorone (M9) and 4,5-dihydro-17alpha-deacetyl-Nestorone (M19) were identified by HPLC and LC/MS analysis. Metabolite profiling of plasma samples showed that most of the circulating radioactivity was associated with unchanged parent drug, and M19. The M19 was a major metabolite in the profiled urine and feces samples. Presence of large proportion of drug/drug-related material in feces suggested that the biliary excretion is a main elimination route of (3)H Nestorone. The distribution, metabolism, and excretion profiles of (3)H Nestorone obtained in this study provide a fairly good insight about its fate in women.


Characterization of a PR-10 pathogenesis-related gene family induced in rice during infection with Magnaporthe grisea.
A partial cDNA with homology to the PR-10 class of pathogenesis-related proteins was used to screen a rice genomic library. One 16-kb genomic clone contained three genes with PR-10 similarity. These genes, RPR10a, RPR10b, and RPR10c, were arranged in tandem and separated by approximately 2.5 kb. RPR10a cDNA was obtained by reverse transcription-polymerase chain reaction, and sequence analysis revealed that RPR10a and RPR10b encode predicted proteins of 158 and 160 amino acids, respectively, and share 71% amino acid identity. RPR10c appears to be a nonfunctional pseudogene. Gene-specific probes were used to study transcript accumulations of the three RPR10 genes in rice plants following inoculation with Magnaporthe grisea. RPR10a transcripts were induced from a low basal level within 12 h after inoculation and showed a second higher level induction at 48 h, which continued throughout the 144 h it was examined. In addition, RPR10a was induced strongly by salicylic and jasmonic acid applications to rice plants. Transcripts of RPR10b also were enhanced by M. grisea, but were not strongly visible until 48 h after inoculation. Tissue prints of M. grisea-infected rice leaves when the RPR10a-specific probe was used indicate that RPR10a is expressed most strongly in a localized fashion in response to the pathogen.


Context matters: the benefits and costs of expressing positive emotion among survivors of childhood sexual abuse.
Positive emotions promote adjustment to aversive life events. However, evolutionary theory and empirical research on trauma disclosure suggest that in the context of stigmatized events, expressing positive emotions might incur social costs. To test this thesis, the authors coded genuine (Duchenne) smiling and laughter and also non-Duchenne smiling from videotapes of late-adolescent and young adult women, approximately half with documented histories of childhood sexual abuse (CSA), as they described the most distressing event of their lives. Consistent with previous studies, genuine positive emotional expression was generally associated with better social adjustment two years later. However, as anticipated, CSA survivors who expressed positive emotion in the context of describing a past CSA experience had poorer long-term social adjustment, whereas CSA survivors who expressed positive emotion while describing a nonabuse experience had improved social adjustment. These findings suggest that the benefits of positive emotional expression may often be context specific.


A quantitative analysis of the structure of human sternum.
An extensive study of the human sternum has been carried out to obtain estimates of the omnidirectional path-length distributions and structural parameters for trabeculation and marrow spaces. Data for sternum samples have been collected, using an object plane scanning microscope. These data have been used to produce the omnidirectional path-length distributions and values of structural parameters for the whole sternum. For a typical adult man the mean trabecular and marrow space path lengths are 224 and 1364 mum, respectively. The percentage bone volume is 13.8 and the surface to volume ratio is 190 cm. Data on the structural variations within the whole sternum are presented. They show a percentage difference in bone volume between the manubrium and the body of sternum of about 36%.


Variants in CUL4B are associated with cerebral malformations.
Variants in cullin 4B (CUL4B) are a known cause of syndromic X-linked intellectual disability. Here, we describe an additional 25 patients from 11 families with variants in CUL4B. We identified nine different novel variants in these families and confirmed the pathogenicity of all nontruncating variants. Neuroimaging data, available for 15 patients, showed the presence of cerebral malformations in ten patients. The cerebral anomalies comprised malformations of cortical development (MCD), ventriculomegaly, and diminished white matter volume. The phenotypic heterogeneity of the cerebral malformations might result from the involvement of CUL-4B in various cellular pathways essential for normal brain development. Accordingly, we show that CUL-4B interacts with WDR62, a protein in which variants were previously identified in patients with microcephaly and a wide range of MCD. This interaction might contribute to the development of cerebral malformations in patients with variants in CUL4B.


Ambiguity: A new way of thinking about responses to climate change.
Diversity, interdisciplinarity and transdisciplinarity are now recognized as vital to tackling wicked problems such as those presented by a changing climate (Nature editorial 2015, Ledford 2015; Dick et al., 2016). Including diverse disciplines in science projects enables a range of different views which often facilitate the creation of innovative solutions. Supporting multiple views and options requires a different way of working beyond traditional reductionist approaches to science, communication and decision-making. To embrace diversity in scientific project teams in order to tackle complex, integrated and urgent issues but to expect singular and linear pathways forward is paradoxical. Much has been written about the need for the scientific community to embrace uncertainty (e.g. Popper, Lempert & Bankes 2005; Lempert et al., 2004; Nelson, Howden & Hayman 2013; Bammer & Smithson 2008). We argue that this in itself will not suffice, and that there is also a need to embrace ambiguity in certain situations. Thus, in this article we explore: (1) what ambiguity is, including the benefits it can offer to climate adaptation in particular, using existing approaches to ambiguity in the arts and humanities as examples (2), we discuss practical meanings of ambiguity in relation to climate change, (3) we propose possible next steps for bringing ambiguity into interdisciplinary practice, and (4) we identify some challenges and necessary preconditions to successfully and appropriately embracing ambiguity.


[Efficacy of fenoverine and trimebutine in the management of irritable bowel syndrome: multicenter randomized double-blind non-inferiority clinical study].
Antispasmodic agents have been used in the management of irritable bowel syndrome. However, systematic reviews have come to different conclusions about the efficacy in irritable bowel syndrome. Fenoverine acts as a synchronizer of smooth muscle in modulating the intracellular influx of calcium. We compared fenoverine with trimebutine for the treatment of patients with IBS. A multicenter, randomized, double-blind, non-inferiority clinical study was conducted to compared fenoverine with trimebutine. Subjects were randomized to receive either fenoverine (100 mg three times a day) or trimebutine (150 mg three times a day) for 8 weeks. A total of 197 patients were analyzed by the intention-to-treat approach. The primary endpoint was the proportion of patients who had 30% reduction in abdominal pain or discomfort measured by bowel symptom scale (BSS) score at week 8 compared to the baseline. The secondary endpoints were changes of abdominal bloating, diarrhea, constipation, overall and total scores of BSS, and overall satisfaction. At week 8, fenoverine was shown to be non-inferior to trimebutine (treatment difference, 1.76%; 90% CI, -10.30-13.82; p=0.81); 69.23% (54 of 78 patients) of patients taking fenoverine and 67.47% (56 of 83 patients) of patients taking trimebutine showed 30% reduction in abdominal pain or discomfort compared to the baseline. There results of the secondary endpoints were also comparable between the fenoverine group and the trimebutine group. Fenoverine is non-inferior to trimebutine for treating IBS in terms of both efficacy and tolerability.


MHC class II epitope nesting modulates dendritic cell function and improves generation of antigen-specific CD4 helper T cells.
CD4 Th cells are critical to the development of coordinated immune responses to infections and tumors. Th cells are activated through interactions of the TCR with MHC class II complexed with peptide. T cell activation is dependent on the density of MHC peptide complexes as well as the duration of interaction of the TCR with APCs. In this study, we sought to determine whether MHC class II peptides could be modified with amino acid sequences that facilitated uptake and presentation with the goal of improving Th cell activation in vitro and in vivo. A model epitope derived from the murine folate receptor α, a self- and tumor Ag, was modified at its carboxyl terminus with the invariant chain-derived Ii-Key peptide and at its N terminus with a peptide that enhances uptake of Ag by APC. Modification of a peptide resulted in enhanced generation of high-avidity murine folate receptor α T cells that persisted in vivo and homed to sites of Ag deposition. The nesting approach was epitope and species independent and specifically excluded expansion of CD4 regulatory T cells. The resulting Th cells were therapeutic, enhanced in vivo helper activity and had an increased ability to resist tolerizing immune microenvironments. In addition to improved immunoadjuvants, this epitope modification strategy may be useful for enhancing ex vivo and in vivo generation of Th cells for preventing and treating diseases.


Comparative analysis of processed ribosomal protein pseudogenes in four mammalian genomes.
The availability of genome sequences of numerous organisms allows comparative study of pseudogenes in syntenic regions. Conservation of pseudogenes suggests that they might have a functional role in some instances. We report the first large-scale comparative analysis of ribosomal protein pseudogenes in four mammalian genomes (human, chimpanzee, mouse and rat). To this end, we have assigned these pseudogenes in the four organisms using an automated pipeline and make the results available online. Each organism has a large number of ribosomal protein pseudogenes (approximately 1,400 to 2,800). The majority of them are processed (generated by retrotransposition). However, we do not see a correlation between the number of pseudogenes associated with a ribosomal protein gene and its mRNA abundance. Analysis of pseudogenes in syntenic regions between species shows that most are conserved between human and chimpanzee, but very few are conserved between primates and rodents. Interestingly, syntenic pseudogenes have a lower rate of nucleotide substitution than their surrounding intergenic DNA. Moreover, evidence from expressed sequence tags indicates that two pseudogenes conserved between human and mouse are transcribed. Detailed analysis shows that one of them, the pseudogene of RPS27, is likely to be a protein-coding gene. This is significant as previous reports indicated there are exactly 80 ribosomal protein genes encoded by the human genome. Our analysis indicates that processed ribosomal protein pseudogenes abound in mammalian genomes, but few of these are conserved between primates and rodents. This highlights the large amount of recent retrotranspositional activity in mammals and a relatively larger amount of it in the rodent lineage.


Melanoma brain metastasis is independent of lactate dehydrogenase A expression.
The key metabolic enzyme lactate dehydrogenase A (LDHA) is overexpressed in many cancers, and several preclinical studies have shown encouraging results of targeted inhibition. However, the mechanistic importance of LDHA in melanoma is largely unknown and hitherto unexplored in brain metastasis. We investigated the spatial, temporal, and functional features of LDHA expression in melanoma brain metastasis across multiple in vitro assays, in a robust and predictive animal model employing MRI and PET imaging, and in a unique cohort of 80 operated patients. We further assessed the genomic and proteomic landscapes of LDHA in different cancers, particularly melanomas. LDHA expression was especially strong in early and small brain metastases in vivo and related to intratumoral hypoxia in late and large brain metastases in vivo and in patients. However, LDHA expression in human brain metastases was not associated with the number of tumors, BRAF(V600E) status, or survival. Moreover, LDHA depletion by small hairpin RNA interference did not affect cell proliferation or 3D tumorsphere growth in vitro or brain metastasis formation or survival in vivo. Integrated analyses of the genomic and proteomic landscapes of LDHA indicated that LDHA is present but not imperative for tumor progression within the CNS, or predictive of survival in melanoma patients. In a large patient cohort and in a robust animal model, we show that although LDHA expression varies biphasically during melanoma brain metastasis formation, tumor progression and survival seem to be functionally independent of LDHA.


Erratum.
In the abstract and in the text (line 35 in the middle column of page 185) of the report "Electrical resistivity and stoichiometry of K(chi)C(60) films" by G. P. Kochanski et al. (10 Jan., p. 184), the minimum resistivity was given incorrectly as 2.2 microohm-cm. The correct value is 2.2 milliohm-cm.


A subtype of childhood acute lymphoblastic leukaemia with poor treatment outcome: a genome-wide classification study.
Genetic subtypes of acute lymphoblastic leukaemia (ALL) are used to determine risk and treatment in children. 25% of precursor B-ALL cases are genetically unclassified and have intermediate prognosis. We aimed to use a genome-wide study to improve prognostic classification of ALL in children. We constructed a classifier based on gene expression in 190 children with newly diagnosed ALL (German Cooperative ALL [COALL] discovery cohort) by use of double-loop cross-validation and validated this in an independent cohort of 107 newly diagnosed patients (Dutch Childhood Oncology Group [DCOG] independent validation cohort). Hierarchical cluster analysis with classifying gene-probe sets revealed a new ALL subtype, the underlying genetic abnormalities of which were characterised by comparative genomic hybridisation-arrays and molecular cytogenetics. Our classifier predicted ALL subtype with a median accuracy of 90.0% (IQR 88.3-91.7) in the discovery cohort and correctly identified 94 of 107 patients (accuracy 87.9%) in the independent validation cohort. Without our classifier, 44 children in the COALL cohort and 33 children in the DCOG cohort would have been classified as B-other. However, hierarchical clustering showed that many of these genetically unclassified cases clustered with BCR-ABL1-positive cases: 30 (19%) of 154 children with precursor B-ALL in the COALL cohort and 14 (15%) of 92 children with precursor B-ALL in the DCOG cohort had this BCR-ABL1-like disease. In the COALL cohort, these patients had unfavourable outcome (5-year disease-free survival 59.5%, 95% CI 37.1-81.9) compared with patients with other precursor B-ALL (84.4%, 76.8-92.1%; p=0.012), a prognosis similar to that of patients with BCR-ABL1-positive ALL (51.9%, 23.1-80.6%). In the DCOG cohort, the prognosis of BCR-ABL1-like disease (57.1%, 31.2-83.1%) was worse than that of other precursor B-ALL (79.2%, 70.2-88.3%; p=0.026), and similar to that of BCR-ABL1-positive ALL (32.5%, 2.3-62.7%). 36 (82%) of the patients with BCR-ABL1-like disease had deletions in genes involved in B-cell development, including IKZF1, TCF3, EBF1, PAX5, and VPREB1; only nine (36%) of 25 patients with B-other ALL had deletions in these genes (p=0.0002). Compared with other precursor B-ALL cells, BCR-ABL1-like cells were 73 times more resistant to L-asparaginase (p=0.001) and 1.6 times more resistant to daunorubicin (p=0.017), but toxicity of prednisolone and vincristine did not differ. New treatment strategies are needed to improve outcome for this newly identified high-risk subtype of ALL. Dutch Cancer Society, Sophia Foundation for Medical Research, Paediatric Oncology Foundation Rotterdam, Centre of Medical Systems Biology of the Netherlands Genomics Initiative/Netherlands Organisation for Scientific Research, American National Institute of Health, American National Cancer Institute, and American Lebanese Syrian Associated Charities.


A genome-wide association meta-analysis implicates Hedgehog and Notch signaling in Dupuytren's disease.
Dupuytren's disease (DD) is a highly heritable fibrotic disorder of the hand with incompletely understood etiology. A number of genetic loci, including Wnt signaling members, have been previously identified. Our overall aim was to identify novel genetic loci, to prioritize genes within the loci for functional studies, and to assess genetic correlation with associated disorders. We performed a meta-analysis of six DD genome-wide association studies from three European countries and extensive bioinformatic follow-up analyses. Leveraging 11,320 cases and 47,023 controls, we identified 85 genome-wide significant single nucleotide polymorphisms in 56 loci, of which 11 were novel, explaining 13.3-38.1% of disease variance. Gene prioritization implicated the Hedgehog and Notch signaling pathways. We also identified a significant genetic correlation with frozen shoulder. The pathways identified highlight the potential for new therapeutic targets and provide a basis for additional mechanistic studies for a common disorder that can severely impact hand function.


Mry, a trans-acting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems.
In the Streptococcus pyogenes M6 strain D471, an insertion of the conjugative transposon Tn916 into a region 2 kb upstream of the promoter of emm6 (the structural gene for the M protein) rendered the strain M negative (M. G. Caparon and J. R. Scott, Proc. Natl. Acad. Sci. USA 84:8677-8681, 1987). In the present work, we show that this insertion mutation, mry-1, is 244 bp upstream of an open reading frame encoding a protein we call Mry. This protein is visible on a gel after transcription and translation in vitro. We have developed a technique for complementation analysis in S. pyogenes and have used it to show that the wild-type mry gene is dominant to two mutant alleles. This dominance indicates that Mry acts in trans as a positive regulator of the emm6 gene. The translated DNA sequence of mry has two regions of similarity to the motif common to the receptor protein of two-component regulatory systems. In addition, the N terminus of Mry has two regions resembling a helix-turn-helix motif. Mry does not appear to be a global regulator of virulence determinants in the group A streptococcus because there is no effect of the mry-1 mutation on production of the hyaluronic acid capsule or streptokinase.


Effects of dexamethasone-21-isonicotinate on peripheral insulin action in dairy cows 5 days after surgical correction of abomasal displacement.
Dexamethasone frequently is used for treatment of ketosis in dairy cows, but its effects are not fully understood. Dexamethasone treatment affects whole body insulin sensitivity. Twelve German Holstein cows, 2-4 weeks postpartum, 5 days after omentopexy to correct left abomasal displacement. Randomized, blinded, case-control study. Treatment with dexamethasone-21-isonicotinate (DG; 40 μg/kg IM; n = 6) or saline (control group [CG], 15 mL IM, n = 6) on day 0 (d0). Blood samples were obtained before (d0) and after treatment (d1 and d2), and analyzed for glucose, insulin, and nonesterified fatty acid (NEFA) concentrations. Hepatic triglycerides (TAG) were measured in liver samples taken on d0 and d2. Five consecutive hyperinsulinemic-euglycemic clamps (HEC-I-V; insulin dosages: 0.1, 0.5, 2, 5, 10 mU/kg/min, respectively) were performed on d1 and steady state glucose infusion rate (SSGIR), insulin concentration (SSIC), insulin sensitivity index (ISI = SSGIR/SSIC), and plasma NEFA concentration (SSNEFA) were assessed. Compared with CG-cows, DG-cows on d1 had higher plasma glucose (P = .004) and insulin (P < .001) concentrations, decreased SSGIR (HEC-II, P = .002; HEC-IV, P = .033), ISI (HEC-I, P < .015; HEC-II, P = .004), and insulin-stimulated decrease in SSNEFA (HEC-II, P = .006; HEC-III, P = .01; HEC-IV, P = .003; HEC-V, P = .011). Decrease in hepatic TAG content in DG-cows was higher compared with CG-cows (P < .001). Dexamethasone decreases whole body insulin sensitivity and affects glucose and lipid metabolism in early lactating dairy cows.


Molecular mapping of the RNA Cap 2'-O-methyltransferase activation interface between severe acute respiratory syndrome coronavirus nsp10 and nsp16.
Several protein-protein interactions within the SARS-CoV proteome have been identified, one of them being between non-structural proteins nsp10 and nsp16. In this work, we have mapped key residues on the nsp10 surface involved in this interaction. Alanine-scanning mutagenesis, bioinformatics, and molecular modeling were used to identify several "hot spots," such as Val(42), Met(44), Ala(71), Lys(93), Gly(94), and Tyr(96), forming a continuous protein-protein surface of about 830 Å(2), bearing very conserved amino acids among coronaviruses. Because nsp16 carries RNA cap 2'-O-methyltransferase (2'O-MTase) activity only in the presence of its interacting partner nsp10 (Bouvet, M., Debarnot, C., Imbert, I., Selisko, B., Snijder, E. J., Canard, B., and Decroly, E. (2010) PLoS Pathog. 6, e1000863), functional consequences of mutations on this surface were evaluated biochemically. Most changes that disrupted the nsp10-nsp16 interaction without structural perturbations were shown to abrogate stimulation of nsp16 RNA cap 2'O-MTase activity. More strikingly, the Y96A mutation abrogates stimulation of nsp16 2'O-MTase activity, whereas Y96F overstimulates it. Thus, the nsp10-nsp16 interface may represent an attractive target for antivirals against human and animal pathogenic coronaviruses.


Effects of curcumin on bladder cancer cells and development of urothelial tumors in a rat bladder carcinogenesis model.
Curcumin, a well-known dietary pigment derived from Curcuma longa, inhibited growth of several types of malignant cells both in vivo and in vitro. Its effects on cell proliferation and the induction of apoptosis in human bladder cancer cell lines and intravesical activity in a rat bladder tumor model were studied. Exposure of human bladder cancer cells to curcumin resulted in the induction of apoptotic cell death and caused cells to arrest in the G2/M phase. The anti-apoptotic Bcl-2 and Survivin protein was downregulated by the curcumin treatment together with enhancement of the Bax and p53 expression. The inhibitory activities of curcumin were stronger than those of cisplatin and could not be prevented by catalase pretreatment in T24 cells. Clonal assay indicated large-dose and short-term curcumin was lethal to bladder cancer cells. Moreover, the in vivo study revealed curcumin did induce apoptosis in situ, inhibit and slow the development of bladder cancer. These observations suggest that curcumin could prove an effective chemopreventive and chemotherapy agent for bladder cancer.


Three-dimensionally plotted MBG/PHBHHx composite scaffold for antitubercular drug delivery and tissue regeneration.
A suitable drug-loaded scaffold that can postoperatively release an antituberculosis drug efficiently in a lesion area and help repair a bone defect is very important in the clinical treatment of bone tuberculosis (TB). In this study, a composite drug-loaded cylindrical scaffold was prepared by using three-dimensional printing technology in combination with the mesoporous confinement range, surface chemical groups, and gradual degradation of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate). This achieves the slow release of a drug for as long as possible. We implanted the drug-loaded compound scaffold into New Zealand rabbits' femur defect model to study the in vivo drug release performance and osteogenic ability. The in vivo release of isoniazid and rifampicin from the prepared composites could be effectively sustained for 12 weeks in local tissues, whereas these drugs were sustained for just 2 weeks in a control group. The blood drug concentrations were very low and most concentrations were below 5 μg/ml. Therefore, the systemic toxic adverse effect is very low. In addition, the composite exhibits good osteogenic potential in a rabbit bone defect model. The results of this study indicate that this composite has great potential for treating osteoarticular TB.


Does the extracorporeal blood flow affect survival of the arteriovenous vascular access?
Hemodiafiltration with high-convective volumes is associated with improved patient survival, whereby practical realization is contingent on high extracorporeal blood flow (Qb) and dialysis treatment time. However, Qb is restricted by vascular access (VA) quality and/or concerns that high Qb could damage the VA. Taking VA quality into consideration, one can investigate the relationship between Qb and VA survival. We analyzed data from 1039 patients treated by hemodiafiltration over a 21-month period where access blood flow (Qa) measurements were also available at baseline. VA failure was defined as a surgical intervention resulting in the generation of a new VA. Qa was included as a stratification variable within a Cox regression model. A second Cox proportional hazard model with a penalized spline was used to describe the association between Qb and VA survival. Compared with Qb in the 350-357 mL/min range, a significantly higher hazard ratio (HR) for VA failure was detected for fistula only, and then only for Qb < 312 mL/min (HR: 2.361, 95% confidence interval [CI]: 1.251-4.453), Qb = 387-397 mL/min (HR: 1.920, 95% CI: 1.007-3.660) and Qb >414 mL/min (HR: 2.207, 95% CI: 1.101-4.424). Age, gender, diabetes, VA vintage, position of the VA, and arterial pressure were not significantly associated with outcome. The form of the penalized spline confirmed higher risk for VA failure for the lowest and the highest values of Qb. Taking Qa into consideration, no association was found between VA failure and Qb up to flows as high as approximately 390 mL/min.


HLA-B27 predicts a more chronic disease course in an 8-year followup cohort of patients with juvenile idiopathic arthritis.
We investigated associations of HLA-B27 with clinical manifestations and longterm outcome in a near population-based setting among patients with juvenile idiopathic arthritis (JIA). We studied clinical and serological data from 410 patients with HLA-B27 results among 440 prospectively collected patients with JIA with 8-year followup data in a Nordic database. The study was structured to be as close to a population-based study as possible. HLA-B27 was analyzed in 93% of patients, and was positive in 21% of the cohort, in 18.4% of the girls and in 25.9% of the boys. Boys who were HLA-B27-positive had significantly higher age at onset compared to HLA-B27-negative boys and compared to both HLA-B27-negative and positive girls. This difference in onset age in relation to HLA-B27 was not found in girls. HLA-B27 was associated with clinical signs of sacroiliitis, enthesitis, and tenosynovitis in boys, but not in girls. After 8 years of disease, 46 children (11.2%) were classified as having enthesitis-related arthritis (ERA). Boys with ERA had clinical signs of sacroiliitis more often than girls with ERA. HLA-B27-positive children, as well as children with clinical signs of sacroiliitis, enthesitis, and hip arthritis, had higher odds of not being in remission off medication after 8 years of disease. In this near population-based Nordic JIA cohort we found significant differences between HLA-B27-positive boys and girls in age at disease onset, clinical signs of sacroiliitis, and ERA classification. HLA-B27 was negatively associated with longterm remission status, possibly because of its association with clinical disease characteristics, such as sacroiliitis, rather than being a general marker of persistent disease.


Intramedullary screw fixation of Jones fractures.
Jones fractures of the fifth metatarsal can be stabilized using intramedullary screw fixation techniques. A range of screw diameters from 4.5 mm to 6.5 mm can be used, but the optimal screw for this procedure has yet to be defined. In clinical practice, we have observed that failure is more likely when smaller diameter screws are used. Experimental Jones fractures were created in 23 pairs of human cadaver fifth metatarsals, which were fixed using either 5.0 mm or 6.5 mm screws. Fracture stiffness and pull-out strengths were measured for either screw type and their relationships with bone mineral density and medullary canal diameter were determined. There was no significant difference in the bending stiffness of fractures stabilized with 5.0 mm and 6.5 mm screws; however, different mechanisms of failure were noted for either screw type. Poor thread purchase within the medullary canal was noted with the 5.0 mm screws, while excellent purchase was noted with 6.5 mm screws. Pull-out strength testing revealed significantly higher pullout strengths for the larger 6.5 mm screws. There was no significant difference in bone mineral density or medullary canal diameter between right and left metatarsals. Fifth metatarsals can often accommodate a 6.5 mm screw for the stabilization of Jones fractures. Larger diameter screws did not result in greater fracture stiffness in our model, but did result in significantly greater pull-out strengths. Larger diameter screws may be more appropriate for intramedullary screw fixation of Jones fractures.


Simple high-throughput analytical method using ultra-performance liquid chromatography coupled with tandem mass spectrometry to quantify total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol in urine.
Since the urinary concentration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a reliable biomarker of exposure to tobacco smoke, we developed a relatively simple high-throughput chromatographic method to quantify total urinary NNAL concentrations in the general population. The high-throughput analytical method was developed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) to identify and quantify total urinary NNAL concentrations in 10 non-smokers and 15 otherwise healthy smokers. Loss of nitric oxide at m/z 30 was found to be the predominant mass transitioned, and therefore was used as the SIM transition to quantify both NNAL and NNAL-methyl-d3 in urine. The analytical method did not require sample derivatization. Standard curves for total NNAL concentrations were linear between 20 and 1500 pg/mL, with coefficients of determination >0.95. Precision and accuracy ranged from 2.2% to 8.6% (CV) and from -5.6% to 10.9% (percent error), respectively. The lowest limit of quantification was 6.7 pg/mL, and 2.0 pg/mL the lowest limit of detection (LLOD). Total urinary NNAL concentrations in non-smoker subjects were <LLOD, whereas in smokers varied between <LLOD to 112.1 pg/mL. An UPLC-MS/MS analytical method to quantify total urinary NNAL concentrations in smokers that does not require sample derivatization is presented herein. The method could be useful in clarifying the toxicities associated with human exposure to cigarette smoking. However, quantification might be adversely affected by co-eluting interfering compounds or selective ion suppression or enhancement as a result of having only one ion transition to monitor NNAL and NNAL-methyl-d3 in urine.


Perfusion cell seeding on large porous PLA/calcium phosphate composite scaffolds in a perfusion bioreactor system under varying perfusion parameters.
A promising approach to bone tissue engineering lies in the use of perfusion bioreactors where cells are seeded and cultured on scaffolds under conditions of enhanced nutrient supply and removal of metabolic products. Fluid flow alterations can stimulate cell activity, making the engineering of tissue more efficient. Most bioreactor systems are used to culture cells on thin scaffold discs. In clinical use, however, bone substitutes of large dimensions are needed. In this study, MG63 osteoblast-like cells were seeded on large porous PLA/glass scaffolds with a custom developed perfusion bioreactor system. Cells were seeded by oscillating perfusion of cell suspension through the scaffolds. Applicable perfusion parameters for successful cell seeding were determined by varying fluid flow velocity and perfusion cycle number. After perfusion, cell seeding, the cell distribution, and cell seeding efficiency were determined. A fluid flow velocity of 5 mm/s had to be exceeded to achieve a uniform cell distribution throughout the scaffold interior. Cell seeding efficiencies of up to 50% were achieved. Results suggested that perfusion cycle number influenced cell seeding efficiency rather than fluid flow velocities. The cell seeding conducted is a promising basis for further long term cell culture studies in large porous scaffolds.


Atom tunnelling in the reaction NH3+ + H2 → NH4+ + H and its astrochemical relevance.
The title reaction is involved in the formation of ammonia in the interstellar medium. We have calculated thermal rates including atom tunnelling using different rate theories. Canonical variational theory with microcanonically optimised multidimensional tunnelling was used for bimolecular rates, modelling the gas-phase reaction and also a surface-catalysed reaction of the Eley-Rideal type. Instanton theory provided unimolecular rates, which model the Langmuir-Hinshelwood type surface reaction. The potential energy was calculated on the CCSD(T)-F12 level of theory on the fly. We report thermal rates and H/D kinetic isotope effects. The latter have implications for observed H/D fractionation in molecular clouds. Tunnelling causes rate constants to be sufficient for the reaction to play a role in interstellar chemistry even at cryogenic temperature. We also discuss intricacies and limitations of the different tunnelling approximations to treat this reaction, including its pre-reactive minimum.


The efficacy of treadmill training on balance dysfunction in individuals with chronic stroke: a systematic review.
Physical activity and exercise interventions are useful in facilitating the functional recovery of those with chronic stroke and, routinely, are gait-specific. While treadmill training has proven useful in gait performance recovery post-stroke, its efficacy on balance dysfunction has not been systematically reviewed. The purpose of this systematic review was to determine the effect of treadmill training (TT) interventions on balance dysfunction in individuals with chronic stroke. A systematic literature search of PubMed, EMBASE, and CINAHL was performed. Eligible randomized controlled trials were published between 2007 and 2016. Selected trials investigated TT interventions in persons with chronic stroke and implemented at least one objective balance measure. Methodological quality was assessed using PEDro criteria. Eight studies met eligibility criteria and were included in the qualitative analysis. Studies differed in TT implementation and use of adjunctive treatments; however, all trials demonstrated improvements in balance measures that were as effective, if not more so, than conventional physical therapy treatments, including targeted balance training. This review recognized moderate evidence in favor of TT interventions in balance and stroke rehabilitation programs. With TT, intensity may be a more critical factor than specificity and may offer additional carryover to recovery parameters of postural control and balance, beyond gait performance. It is recommended that clinicians utilizing TT incorporate objective measures of balance to assess the potential for skill transference and improvements in balance. Higher quality studies and additional research are needed to denote critical parameters by which improvements in balance may be optimized.


Stress molecules in sepsis and systemic inflammatory response syndrome.
During sepsis, microbial derived products ("pathogen-associated molecular patterns", PAMPs) are recognized as exogenous danger signals by specific sensors of the host ("pattern recognitions receptors", PRRs). This interaction leads to the release of numerous stress proteins that are a prerequisite to fight infection, though their overzealous production can contribute to tissue damage, organ dysfunction and eventually death. In critically ill patients, translocation of PAMPs can occur from the gut, and injured tissues and cells release endogenous danger signals called "alarmins" (e.g. High mobility group box-1); that share some properties with PAMPs. Thus, numerous similarities occur during infectious and non-infectious systemic inflammation.


Vestibular and visual climbing fiber signals evoked in the uvula-nodulus of the rabbit cerebellum by natural stimulation.
1. The cerebellar uvula-nodulus receives vestibular projections from primary and secondary vestibular afferents as well as vestibularly related climbing fibers. It also receives visually related information from climbing fiber pathways. In this experiment we investigated how this information is mapped onto the uvula-nodulus. We studied the specificity, dynamics, and topographic distribution of climbing fiber responses (CFRs), simple spike responses, and mossy fiber terminal responses evoked by vestibular and optokinetic stimulation in rabbits anesthetized with alpha-chloralose. 2. Vestibularly evoked CFRs were found in the ventral uvula and nodulus. These responses were evoked during static roll tilt of the rabbit about a longitudinal axis and by sinusoidal oscillation about the longitudinal axis. Purely static responses were attributed to stimulation of the utricular otolith by the linear acceleration of gravity. CFRs that lacked a static component were attributed to activation of the semicircular canals. 3. Using a "null technique" we showed that the canal-sensitive CFRs were caused by stimulation of the anterior or posterior semicircular canals. Of the CFRs classified as canal related, 96% could be attributed to stimulation of the vertical semicircular canals. 4. Increases in CFRs were correlated with decreases in simple spike responses in half the Purkinje cells from which we recorded. These climbing-fiber-induced pauses in simple spikes occurred during spontaneous climbing fiber discharge as well as during climbing fiber discharge evoked by vestibular stimulation. The duration of this pause was inversely proportional to the spontaneous level of simple spikes before the occurrence of a CFR. In the other half of the recorded population of Purkinje cells, vestibularly driven CFRs did not alter the simple spike responses. 5. Vestibularly and visually mediated CFRs were topographically represented on the surface of the uvula-nodulus. CFRs driven by ipsilateral otolithic inputs were distributed over the entire mediolateral surface of the uvula-nodulus. CFRs driven by the ipsilateral posterior semicircular canal were distributed in a sagittal strip approximately 1.5 mm wide, extending laterally from the midline of the nodulus. CFRs driven exclusively by horizontal, posterior-->anterior optokinetic stimulation of the ipsilateral eye were distributed in a sagittal strip approximately 0.5 mm wide located 0.5-1.0 mm from the midline and restricted to the ventral nodulus. CFRs driven by the ipsilateral anterior semicircular canal were found in a sagittal strip approximately 1.0 mm wide extending 1.0-2.0 mm from the midline. 6. The sagittal, topographically arrayed climbing fiber strips effectively map a mediolateral gradient of possible postural responses based on vestibular and optokinetic information.


Salivette, a relevant saliva sampling device for SARS-CoV-2 detection.
Background: The gold standard for COVID-19 diagnosis relies on quantitative reverse-transcriptase polymerase-chain reaction (RT-qPCR) from nasopharyngeal swab (NPS) specimens, but NPSs present several limitations. The simplicity, low invasive and possibility of self-collection of saliva imposed these specimens as a relevant alternative for SARS-CoV-2 detection. However, the discrepancy of saliva test results compared to NPSs made of its use controversial. Here, we assessed Salivettes®, as a standardized saliva collection device, and compared SARS-CoV-2 positivity on paired NPS and saliva specimens. Methods: A total of 303 individuals randomly selected among those investigated for SARS-CoV-2 were enrolled, including 30 (9.9%) patients previously positively tested using NPS (follow-up group), 90 (29.7%) mildly symptomatic and 183 (60.4%) asymptomatic. Results: The RT-qPCR revealed a positive rate of 11.6% (n = 35) and 17.2% (n = 52) for NPSs and saliva samples, respectively. The sensitivity and specificity of saliva samples were 82.9% and 91.4%, respectively, using NPS as reference. The highest proportion of discordant results concerned the follow-up group (33.3%). Although the agreement exceeded 90.0% in the symptomatic and asymptomatic groups, 17 individuals were detected positive only in saliva samples, with consistent medical arguments. Conclusion Saliva collected with Salivette® was more sensitive for detecting symptomatic and pre-symptomatic infections.


Ecosystem services and urban heat riskscape moderation: water, green spaces, and social inequality in Phoenix, USA.
Urban ecosystems are subjected to high temperatures--extreme heat events, chronically hot weather, or both-through interactions between local and global climate processes. Urban vegetation may provide a cooling ecosystem service, although many knowledge gaps exist in the biophysical and social dynamics of using this service to reduce climate extremes. To better understand patterns of urban vegetated cooling, the potential water requirements to supply these services, and differential access to these services between residential neighborhoods, we evaluated three decades (1970-2000) of land surface characteristics and residential segregation by income in the Phoenix, Arizona, USA metropolitan region. We developed an ecosystem service trade-offs approach to assess the urban heat riskscape, defined as the spatial variation in risk exposure and potential human vulnerability to extreme heat. In this region, vegetation provided nearly a 25 degrees C surface cooling compared to bare soil on low-humidity summer days; the magnitude of this service was strongly coupled to air temperature and vapor pressure deficits. To estimate the water loss associated with land-surface cooling, we applied a surface energy balance model. Our initial estimates suggest 2.7 mm/d of water may be used in supplying cooling ecosystem services in the Phoenix region on a summer day. The availability and corresponding resource use requirements of these ecosystem services had a strongly positive relationship with neighborhood income in the year 2000. However, economic stratification in access to services is a recent development: no vegetation-income relationship was observed in 1970, and a clear trend of increasing correlation was evident through 2000. To alleviate neighborhood inequality in risks from extreme heat through increased vegetation and evaporative cooling, large increases in regional water use would be required. Together, these results suggest the need for a systems evaluation of the benefits, costs, spatial structure, and temporal trajectory for the use of ecosystem services to moderate climate extremes. Increasing vegetation is one strategy for moderating regional climate changes in urban areas and simultaneously providing multiple ecosystem services. However, vegetation has economic, water, and social equity implications that vary dramatically across neighborhoods and need to be managed through informed environmental policies.


Idiotype vaccination using dendritic cells after autologous peripheral blood progenitor cell transplantation for multiple myeloma.
The idiotype (Id) determinants on the multiple myeloma immunoglobulin can serve as tumor-specific antigens. An anti-Id immune response may stem the growth of the malignant clone. We report on 26 patients treated at our institution with high-dose chemotherapy and peripheral blood progenitor cell transplantation (PBPCT) and vaccinated with the Id protein. The patients received chemotherapy and PBPCT to establish a minimal residual disease state. After high-dose therapy, the patients received a series of monthly immunizations consisting of 2 intravenous infusions of dendritic cells (DCs) pulsed with either Id protein or Id coupled with keyhole limpet hemocyanin (KLH) as an immunogenic carrier protein, followed by subcutaneous boosts of Id-KLH conjugates. DCs were obtained in all patients from a leukapheresis product 3 to 9 months after PBPCT. Patients were observed for toxicity, immune responses, and tumor status. The DC infusions and the administration of Id-KLH boosts were well tolerated, with patients experiencing only minor and transient side effects. Of the patients, 24 of 26 generated a KLH-specific cellular proliferative immune response. Only 4 patients developed an Id-specific proliferative immune response. Three of these immune responders were in complete remission at the time of vaccination. A total of 17 patients are alive at a median follow-up of 30 months after transplantation. Id vaccination with autologous DCs is feasible for myeloma patients after transplantation. Id-specific cellular responses can be induced in patients who are in complete remission. Further studies are needed to increase the rate of anti-Id immune responses in patients who do not achieve complete remission.


Chondrosarcoma of bone in children and adolescents.
Chondrosarcomas typically present in adults during the fifth to seventh decades and are rare in young patients. The biological behaviour and oncological outcomes may be different in children and adolescents. We retrospectively evaluated the outcomes of all patients with chondrosarcoma of bone who were younger than 18 years of age at the time of diagnosis and were treated at our centre between 1995 and 2018. The 15 consecutive patients studied included nine male and six female cases, with a mean age at diagnosis of 13 years (7 to 17). The median follow-up was 117 months (30 to 277). The tumours were primary and secondary in ten and five patients, respectively. The tumours were central in 13 and surface in two patients. The tumour locations were the humerus in five, digits in five, femur in three, radius in one and pelvis in one patient. The histological grades were grade I in seven, grade II in seven and grade III in one patient. The surgical treatments were limb salvage in ten patients and ray amputation in five patients. The surgical margins were wide in eight, marginal in two and intralesional in five patients. All the patients were alive and continuously free of disease at the time of the last follow-up. No patient developed metastases or local recurrence. Chondrosarcoma of bone in children and adolescent patients has a very good prognosis and is less aggressive compared with published outcomes in older patients. IV.


Presynaptic Active Zone Plasticity Encodes Sleep Need in Drosophila.
Sleep is universal across species and essential for quality of life and health, as evidenced by the consequences of sleep loss. Sleep might homeostatically normalize synaptic gains made over wake states in order to reset information processing and storage and support learning, and sleep-associated synaptic (ultra)structural changes have been demonstrated recently. However, causal relationships between the molecular and (ultra)structural status of synapses, sleep homeostatic regulation, and learning processes have yet to be established. We show here that the status of the presynaptic active zone can directly control sleep in Drosophila. Short sleep mutants showed a brain-wide upregulation of core presynaptic scaffold proteins and release factors. Increasing the gene copy number of ELKS-family scaffold master organizer Bruchpilot (BRP) not only mimicked changes in the active zone scaffold and release proteins but importantly provoked sleep in a dosage-dependent manner, qualitatively and quantitatively reminiscent of sleep deprivation effects. Conversely, reducing the brp copy number decreased sleep in short sleep mutant backgrounds, suggesting a specific role of the active zone plasticity in homeostatic sleep regulation. Finally, elimination of BRP specifically in the sleep-promoting R2 neurons of 4xBRP animals partially restored sleep patterns and rescued learning deficits. Our results suggest that the presynaptic active zone plasticity driven by BRP operates as a sleep homeostatic actuator that also restricts periods of effective learning.


Growth hormone secretion during sleep.
Plasma growth hormone (GH), insulin, cortisol, and glucose were measured during sleep on 38 nights in eight young adults. Blood was drawn from an indwelling catheter at 30-min intervals; EEG and electrooculogram were recorded throughout the night. In seven subjects, a plasma GH peak (13-72 mmug/ml) lasting 1.5-3.5 hr appeared with the onset of deep sleep. Smaller GH peaks (6-14 mmug/ml) occasionally appeared during subsequent deep sleep phases. Peak GH secretion was delayed if the onset of sleep was delayed. Subjects who were awakened for 2-3 hr and allowed to return to sleep exhibited another peak of GH secretion (14-46 mmug/ml). Peak GH secretion was not correlated with changes in plasma glucose, insulin, and cortisol. The effects of 6-CNS-active drugs on sleep-related GH secretion were investigated. Imipramine (50 mg) completely abolished GH peaks in two of four subjects, whereas chlorpromazine (30 mg), phenobarbital (97 mg), diphenylhydantoin (90 mg), chlordiazepoxide (20 mg), and isocarboxazid (30 mg) did not inhibit GH peaks. Altered hypothalamic activity associated with initiation of sleep results in a major peak of growth hormone secretion unrelated to hypoglycemia or changes in cortisol and insulin secretion.


Adaptation of the Health Literacy Survey Questionnaire (HLS19-Q) for Russian-Speaking Populations-International Collaboration across Germany, Israel, Kazakhstan, Russia, and the USA.
The Russian language is the eighth most spoken language in the world. Russian speakers reside in Russia, across the former Soviet Union republics, and comprise one of the largest populations of international migrants. However, little is known about their health literacy (HL) and there is limited research on HL instruments in the Russian language. The purpose of this study was to adapt the Health Literacy Questionnaire (HLS19-Q) developed within the Health Literacy Survey 2019-2021 (HLS19) to the Russian language to study HL in Russian-speaking populations in Germany, Israel, Kazakhstan, Russia, and the USA. The HLS19-Q was translated either from English or from a national language to Russian in four countries first and then critically reviewed by three Russian-speaking experts for consensus. The HLS19 protocol and "team approach" method were used for linguistic and cultural adaptation. The most challenging was the adaptation of HLS19-Q questions to each country's healthcare system while general HL questions were flexible and adaptable to specific contexts across all countries. This study provides recommendations for the linguistic and cultural adaptation of HLS19-Q into different languages and can serve as an example of international collaboration towards this end.


Point prevalence of bulimia nervosa in 1982, 1992, and 2002.
Background. Recent epidemiological data suggest a decline in bulimia nervosa (BN) incidence in primary care. We sought to examine BN point prevalence from 1982 to 2002 in a college population.Method. In 1982, 1992, and 2002, 800 women and 400 men were randomly sampled from a university for a study of health and eating patterns. Participation rates were 72% in women and 63% in men, resulting in n=2491 participants.Results. BN point prevalence decreased significantly in women over the period of observation. Eating Disorder Inventory Bulimia scores decreased across cohorts, and these decreases remained significant when male and female and Caucasian and non-Caucasian students were analyzed separately.Conclusion. These data support a decline in BN rates that cannot be attributed to changes in service utilization. Changing socio-cultural factors may explain a true decrease in BN incidence and prevalence.


Metrology of replicated diffractive optics with Mueller polarimetry in conical diffraction.
The feasibility of metrological characterization of the one-dimensional (1D) holographic gratings, used in the nanoimprint molding tool fabrication step, by spectroscopic Mueller polarimetry in conical diffraction is investigated. The studied samples correspond to two different steps of the replicated diffraction grating fabrication process. We characterized master gratings that consist of patterned resist layer on chromium-covered glass substrate and complementary (replica) gratings made of nickel. The profiles of the gratings obtained by fitting the experimental spectra of Mueller matrix coefficients taken at different azimuthal angles were confirmed by atomic force microscopy (AFM) measurements. The calculated profiles of corresponding master and replica gratings are found to be complementary. We conclude that the Mueller polarimetry, as a fast and non-contact optical characterization technique, can provide the basis for the metrology of the molding tool fabrication step in the nanoimprint technique.


A2AR Antagonism with CPI-444 Induces Antitumor Responses and Augments Efficacy to Anti-PD-(L)1 and Anti-CTLA-4 in Preclinical Models.
Adenosine signaling through A2A receptors (A2AR) expressed on immune cells suppresses antitumor immunity. CPI-444 is a potent, selective, oral A2AR antagonist. Blockade of A2AR with CPI-444 restored T-cell signaling, IL2, and IFNγ production that were suppressed by adenosine analogues in vitro CPI-444 treatment led to dose-dependent inhibition of tumor growth in multiple syngeneic mouse tumor models. Concentrations of extracellular adenosine in the tumor microenvironment, measured using microdialysis, were approximately 100-150 nmol/L and were higher than corresponding subcutaneous tissue. Combining CPI-444 with anti-PD-L1 or anti-CTLA-4 treatment eliminated tumors in up to 90% of treated mice, including restoration of immune responses in models that incompletely responded to anti-PD-L1 or anti-CTLA-4 monotherapy. Tumor growth was fully inhibited when mice with cleared tumors were later rechallenged, indicating that CPI-444 induced systemic antitumor immune memory. CD8+ T-cell depletion abrogated the efficacy of CPI-444 with and without anti-PD-L1 treatment, demonstrating a role for CD8+ T cells in mediating primary and secondary immune responses. The antitumor efficacy of CPI-444 with and without anti-PD-L1 was associated with increased T-cell activation, a compensatory increase in CD73 expression, and induction of a Th1 gene expression signature consistent with immune activation. These results suggest a broad role for adenosine-mediated immunosuppression in tumors and justify the further evaluation of CPI-444 as a therapeutic agent in patients with solid tumors. Cancer Immunol Res; 6(10); 1136-49. ©2018 AACR.


Comparing effects between music intervention and aromatherapy on anxiety of patients undergoing mechanical ventilation in the intensive care unit: a randomized controlled trial.
Using patient-reported outcomes and physiological indicators to test the effects of music intervention and aromatherapy on reducing anxiety for intensive care unit (ICU) patients undergoing mechanical ventilation. Patients with ICU admission duration >24 h were randomly assigned to a Music intervention group (n = 41), Aromatherapy group (n = 47), or Control group (rest only; n = 44). Each patient in the Music group listened to music; each patient in the Aromatherapy group received lavender essential oil massage on his/her back for 5 min; each patient in the Control group wore noise-canceling headphones. Anxiety was measured using the Chinese version of the Stage-Trait Anxiety Inventory (C-STAI) and the Visual Analogue Scale for Anxiety (VAS-A) at baseline, post-test, and 30-min follow-up. Heart rate, breathing rate, and blood pressure were measured every 10 min from baseline to the 30-min follow-up. The Music group had significantly better post-test VAS-A and C-STAI scores, and had lower heart rate and blood pressure than the Control group. The Aromatherapy group had significantly better VAS-A score and lower heart rate than the Control group. The 30-min follow-up showed that both Music and Aromatherapy groups had lower heart rate and blood pressure than the Control group. Music and aromatherapy interventions were both effective for ICU patients. The effects of music intervention were greater than that of aromatherapy; both interventions maintained the effects for at least 30 min.


Simple model of errors in chirped fiber gratings.
A simple etalon based model is presented to show the origin of the wavelength-dependent ripples in the group delay and phase, and in the intensity of optical signals reflected from chirped fiber gratings. The simplicity of the model allows intuitive understanding of the effects, and quantitative predictions. We derive accurate scaling laws that allow the experimenter to make quantitative connections between the grating writing process parameters and grating performance.


Detection of complete and partial chromosome gains and losses by comparative genomic in situ hybridization.
Comparative genomic in situ hybridization (CGH) provides a new possibility for searching genomes for imbalanced genetic material. Labeled genomic test DNA, prepared from clinical or tumor specimens, is mixed with differently labeled control DNA prepared from cells with normal chromosome complements. The mixed probe is used for chromosomal in situ suppression (CISS) hybridization to normal metaphase spreads (CGH-metaphase spreads). Hybridized test and control DNA sequences are detected via different fluorochromes, e.g., fluorescein isothiocyanate (FITC) and tetraethylrhodamine isothiocyanate (TRITC). The ratios of FITC/TRITC fluorescence intensities for each chromosome or chromosome segment should then reflect its relative copy number in the test genome compared with the control genome, e.g., 0.5 for monosomies, 1 for disomies, 1.5 for trisomies, etc. Initially, model experiments were designed to test the accuracy of fluorescence ratio measurements on single chromosomes. DNAs from up to five human chromosome-specific plasmid libraries were labeled with biotin and digoxigenin in different hapten proportions. Probe mixtures were used for CISS hybridization to normal human metaphase spreads and detected with FITC and TRITC. An epifluorescence microscope equipped with a cooled charge coupled device (CCD) camera was used for image acquisition. Procedures for fluorescence ratio measurements were developed on the basis of commercial image analysis software. For hapten ratios 4/1, 1/1 and 1/4, fluorescence ratio values measured for individual chromosomes could be used as a single reliable parameter for chromosome identification. Our findings indicate (1) a tight correlation of fluorescence ratio values with hapten ratios, and (2) the potential of fluorescence ratio measurements for multiple color chromosome painting. Subsequently, genomic test DNAs, prepared from a patient with Down syndrome, from blood of a patient with T-cell prolymphocytic leukemia, and from cultured cells of a renal papillary carcinoma cell line, were applied in CGH experiments. As expected, significant differences in the fluorescence ratios could be measured for chromosome types present in different copy numbers in these test genomes, including a trisomy of chromosome 21, the smallest autosome of the human complement. In addition, chromosome material involved in partial gains and losses of the different tumors could be mapped to their normal chromosome counterparts in CGH-metaphase spreads. An alternative and simpler evaluation procedure based on visual inspection of CCD images of CGH-metaphase spreads also yielded consistent results from several independent observers. Pitfalls, methodological improvements, and potential applications of CGH analyses are discussed.


Development and validation of a brief pediatric screen for asthma and allergies among children.
Asthma is the most common disease of childhood, but the recognition and detection remain poor, especially among schoolchildren. There has been an increase in the number of instruments available to detect the risk of asthma earlier in children. We have previously validated a simple, self-reported screen, the Brief Pediatric Asthma Screen (BPAS). To develop a new screen for asthma and allergies based on the BPAS (BPAS+) with the intent of keeping the screen brief and simple, while including allergy detection. Questions from the BPAS were extensively revised, and questions regarding allergic rhinitis were added. A panel of parents of asthmatic children reviewed and critiqued the questions. The final BPAS + was distributed in elementary schools, and a cohort of 129 participated in a validation against the gold standard of evaluation by an expert in asthma. For asthma the best items were wheeze, persistent cough, night cough, and response to change in air temperature. The simplest scoring, any 1 of the 4 items, yielded the best balance of specificity (73.6%) and sensitivity (73.3%). For allergy, using all six items, having any one or any two of the items had sensitivity of 71.4% and specificity of 77.3%. The BPAS+ provides a rapid and valid method for the detection of potential allergy and asthma in schoolchildren. Sensitivity and specificity are acceptable for both asthma and allergies.


Analysis and advanced characterization of municipal solid waste vermicompost maturity for a green environment.
Rapid demographic expansion along with increasing urbanization has aggravated the problem of solid waste management. Therefore, scientists are seeking waste management methods that are eco-friendly, cost effective and produce immediate results. In the developing world, municipal solid waste (MSW) contains mostly organic substances, therefore vermicomposting could be a better and cost-effective option for waste management. In this study, vermicomposting of organic portion of MSW with cow dung (additive) was performed using Eisenia fetida. The results showed significant (p < 0.001) decline in pH (13.17%), TOC (21.70%), C: N (62.53%) and C: P (57.66%) ratios, whilst total N (108.9%), P (84.89%) and K (21.85%) content increased (p < 0.001) in matured vermicompost. Different enzymatic activities declined during termination phase of vermicomposting experiment with maximum decrease of 41.72 (p = 0.002) and 39.56% (p = 0.001) in protease and β-glucosidase, respectively. FT-IR, TGA, DSC and SEM studies suggested that final vermicompost was more stabilized as compared to initial waste mixture, characterized by reduced levels of aliphatic materials, carbohydrates and increase in aromatic groups possibly due to biosynthesis of humic substances. Both, the conventional (physicochemical and enzyme activity) and advanced techniques depict maturity and stability of the ready vermicompost. However, FT-IR, TGA, DSC and SEM were proved to be more promising, fast and reliable techniques over conventional analyses.


Heterogeneity in mouse spasmolytic polypeptide-expressing metaplasia lineages identifies markers of metaplastic progression.
Spasmolytic polypeptide-expressing metaplasia (SPEM) develops as a preneoplastic lesion in the stomachs of mice and humans after parietal cell loss. To identify the commonalities and differences between phenotypic SPEM lineages, SPEM were studied from three different mouse models of parietal cell loss: with chronic inflammation with Helicobacter felis infection; with acute inflammation with L635 treatment; and without inflammation following DMP-777 treatment. RNA transcripts from laser capture microdissected normal chief cells and SPEM lineages were compared using gene microarray. Alterations in transcripts were validated by quantitative real-time PCR. Clusterin and cystic fibrosis transmembrane conductance regulator (CFTR) were selected for immunohistochemical analysis in all mouse models as well as in human SPEM, intestinal metaplasia and gastric cancer. Transcript expression patterns demonstrated differences among the phenotypic SPEM models. Clusterin expression was significantly upregulated in all three mouse SPEM models as well as in human SPEM. The highest clusterin expression in human gastric cancers correlated with poor survival. Conversely, CFTR expression was upregulated only in SPEM with inflammation in mice. In humans, intestinal metaplasia, but not SPEM, expressed CFTR. While markers such as clusterin are expressed in all phenotypic SPEM lineages, distinct patterns of upregulated genes including CFTR are present in murine metaplasia associated with inflammation, indicative of progression of metaplasia towards a more intestinalised metaplastic phenotype.


Induction of cytotoxic T lymphocytes against I-region-coded determinants: in vitro evidence for a third histocompatibility locus in the mouse.
Determinants controlled by the I region of the murine H-2 complex provoked the generation of cytotoxic T lymphocytes (CTL) in both a secondary and primary mixed lymphocyte culture. The stimulating determinants appeared to be controlled by loci within the I-A subregion. The target antigens of the CTL generated were present on both lipopolysaccharide- and concanavalin-induced blast lymphocytes, but were barely detectable on phytohemagglutinin-induced blast cells. The stimulating capacity for CTL induction of a complete H-2 complex incompatibility by far exceeded the sum of H-2D/K-region and I-region incompatibility, respectively.