Gene length corrected trimmed mean of M-values (GeTMM) processing of RNA-seq data performs similarly in intersample analyses while improving intrasample comparisons.
Current normalization methods for RNA-sequencing data allow either for intersample comparison to identify differentially expressed (DE) genes or for intrasample comparison for the discovery and validation of gene signatures. Most studies on optimization of normalization methods typically use simulated data to validate methodologies. We describe a new method, GeTMM, which allows for both inter- and intrasample analyses with the same normalized data set. We used actual (i.e. not simulated) RNA-seq data from 263 colon cancers (no biological replicates) and used the same read count data to compare GeTMM with the most commonly used normalization methods (i.e. TMM (used by edgeR), RLE (used by DESeq2) and TPM) with respect to distributions, effect of RNA quality, subtype-classification, recurrence score, recall of DE genes and correlation to RT-qPCR data. We observed a clear benefit for GeTMM and TPM with regard to intrasample comparison while GeTMM performed similar to TMM and RLE normalized data in intersample comparisons. Regarding DE genes, recall was found comparable among the normalization methods, while GeTMM showed the lowest number of false-positive DE genes. Remarkably, we observed limited detrimental effects in samples with low RNA quality. We show that GeTMM outperforms established methods with regard to intrasample comparison while performing equivalent with regard to intersample normalization using the same normalized data. These combined properties enhance the general usefulness of RNA-seq but also the comparability to the many array-based gene expression data in the public domain.


Managing Adverse Events Associated with Dinutuximab Beta Treatment in Patients with High-Risk Neuroblastoma: Practical Guidance.
Neuroblastoma is the most common extracranial solid tumour in children, accounting for 15% of all paediatric cancer deaths. High-risk neuroblastoma is a particularly challenging-to-treat form of disease that requires multimodality treatment, consisting of chemotherapy, surgery, high-dose chemotherapy with autologous haematopoietic stem cell rescue, radiotherapy and differentiation therapy. However, despite intense multimodal treatment regimens, the prognosis for this patient population remains poor. In recent years, immunotherapy with anti-disialoganglioside 2 (anti-GD2) antibodies was found to improve survival rates for patients with high-risk neuroblastoma. Based on studies led by the SIOPEN (International Society of Paediatric Oncology European Neuroblastoma) group, the anti-GD2 antibody dinutuximab beta was approved for use in high-risk neuroblastoma by the European Medicines Agency and has been implemented into the standard of care in many countries across Europe. However, immunotherapy with dinutuximab beta is associated with a number of adverse events that may be challenging for clinicians, such as pain, fever, hypersensitivity reactions and capillary leak syndrome. While these adverse events are considered manageable, there are currently no formal guidelines to support clinicians with their management. The aim of this article is to discuss the management of the most common adverse events encountered in clinical practice and to provide practical guidance to assist clinicians in minimising toxicity associated with dinutuximab beta.


Influence of membrane properties on spike synchronization in neurons: theory and experiments.
In the brain spike synchronization in neurons is involved in information transfer and certain forms of dysfunction. The theory of random point processes was used to relate the statistical properties of input point processes to synchronization between firing in neurons viewed as threshold devices. Derived analytical relations describe normalized synchronization in the case of shared input with balanced excitation and inhibition. For neuronal models with unbalanced shared input and spike generating Hodgkin-Huxley type conductances, the theory satisfactorily describes the temporal dependence of spike synchronization on the delay between spikes. Computer generated stochastic stimulus current was used to stimulate motoneurons in turtle spinal cord slices. Theory was able to approximate the temporal dependence of spike synchronization on the delay between spikes when the membrane time constant and the relative spike threshold measured were used in calculations. In agreement with the theoretical prediction, normalized spike synchrony was reduced when the threshold for spike generation was lowered by injection of steady depolarizing bias current. In spinal motoneurons the relative spike threshold can be lowered by a persistent inward current facilitated by activation of certain metabotropic transmitter receptors. After induction of this inward current spike synchronization was reduced several times. It is suggested that downregulation of the persistent inward current in motoneurons by disruption of brainstem modulatory systems, as in Parkinson disease, can facilitate tremor due to the increased synchrony between motoneurons.


Cortical plasticity induced by transcranial magnetic stimulation during wakefulness affects electroencephalogram activity during sleep.
Sleep electroencephalogram (EEG) brain oscillations in the low-frequency range show local signs of homeostatic regulation after learning. Such increases and decreases of slow wave activity are limited to the cortical regions involved in specific task performance during wakefulness. Here, we test the hypothesis that reorganization of motor cortex produced by long-term potentiation (LTP) affects EEG activity of this brain area during subsequent sleep. By pairing median nerve stimulation with transcranial magnetic stimulation over the contralateral motor cortex, one can potentiate the motor output, which is presumed to reflect plasticity of the neural circuitry. This paired associative stimulation increases M1 cortical excitability at interstimulus intervals of 25 ms. We compared the scalp distribution of sleep EEG power following paired associative stimulation at 25 ms to that following a control paradigm with 50 ms intervals. It is shown that the experimental manipulation by paired associative stimulation at 25 ms induces a 48% increase in amplitude of motor evoked potentials. This LTP-like potentiation, induced during waking, affects delta and theta EEG power in both REM and non-REM sleep, measured during the following night. Slow-wave activity increases in some frontal and prefrontal derivations and decreases at sites neighboring and contralateral to the stimulated motor cortex. The magnitude of increased amplitudes of motor evoked potentials by the paired associative stimulation at 25 ms predicts enhancements of slow-wave activity in prefrontal regions. An LTP-like paradigm, presumably inducing increased synaptic strength, leads to changes in local sleep regulation, as indexed by EEG slow-wave activity. Enhancement and depression of slow-wave activity are interpreted in terms of a simultaneous activation of both excitatory and inhibitory circuits consequent to the paired associative stimulation at 25 ms.


Specific, high affinity binding of tissue inhibitor of metalloproteinases-4 (TIMP-4) to the COOH-terminal hemopexin-like domain of human gelatinase A. TIMP-4 binds progelatinase A and the COOH-terminal domain in a similar manner to TIMP-2.
The binding properties of the newly described tissue inhibitor of metalloproteinases-4 (TIMP-4) to progelatinase A and to the COOH-terminal hemopexin-like domain (C domain) of the enzyme were examined. We present evidence for the first time of a specific, high affinity interaction between TIMP-4 and the C domain of human gelatinase A and show that TIMP-4 binds both progelatinase A and the C domain in a similar manner to that of TIMP-2. Saturable binding of recombinant C domain to TIMP-4 and to TIMP-2 but not to TIMP-1 was demonstrated using a microwell protein binding assay. The recombinant collagen binding domain of gelatinase A, comprised of the three fibronectin type II-like repeats, did not bind to TIMP-4, indicating that binding is mediated selectively by the C domain. Binding to TIMP-4 was of high affinity with an apparent Kd of 1.7 x 10(-7) M but slightly weaker than that to TIMP-2 (apparent Kd of 0.66 x 10(-7) M). Affinity chromatography confirmed the TIMP-4-C domain interaction and also showed that the complex could not be disrupted by 1 M NaCl or 10% dimethyl sulfoxide, thereby further demonstrating the tight binding. To verify the biological significance of this interaction, binding of full-length progelatinase A to TIMP-4 was investigated. TIMP-4 and TIMP-2 but not TIMP-1 bound specifically to purified TIMP-2-free human recombinant full-length progelatinase A and to full-length rat proenzyme from the conditioned culture medium of ROS 17/2.8 cells. Preincubation of the C domain with TIMP-2 was found to reduce subsequent binding to TIMP-4 in a concentration-dependent manner. Competition between TIMP-2 and TIMP-4 for a common or overlapping binding sites on the gelatinase A C domain may occur; alternatively TIMP-2 may prevent the binding of TIMP-4 by steric hindrance or induction of a conformational change in the C domain. We propose that the binding of progelatinase A to TIMP-4 represents a third TIMP-progelatinase interaction in addition to that of progelatinase A with TIMP-2 and progelatinase B with TIMP-1 described previously. This new phenomenon may be of important physiological significance in modulating the cell surface activation of progelatinase A.


Status of liver and gastrointestinal transplantation at the University of Miami.
The liver transplant program at the University of Miami, established in 1987, was rejuvenated in June 1994 with the addition of new staff and expanded to include all organs of the gastrointestinal tract. Since its inception, 630 patients have been transplanted in the program. During the past 2 years we performed 349 transplants in 318 patients (livers n = 323 in 298 patients, liver + kidneys n = 13, liver + islet n = 10, liver + kidney + islets n = 1, liver + heart n = 10, liver + lung n = 1). These included 4 split-liver, 3 living-related, multiple reduced-sized and one "Domino" liver transplant. We have an active pediatric program and 10% of our transplanted patients are pediatric. Our overall patient and graft survival rates were 81% and 78%, respectively. The intestinal transplant program was launched in August 1994. To date we have performed 22 intestinal transplants, in 9 adults and 13 children. These transplants included 4 isolated intestinal, 11 combined liver-intestinal and 7 multivisceral transplants. Overall patient and graft survival rates were 55% and 50%, respectively. During the past 2 years several studies involving immunosuppressive agents were carried out: 1)Mycophenolate Mofetil (MMF) was used as induction therapy and as rescue therapy in patients with steroid-resistant rejection. Tacrolimus toxicity, and chronic rejection; 2) Neoral was compared with Tacrolimus in patients with Hepatitis C; and 3) MMF was added as triple therapy for the intestinal transplants. We used alpha interferon-2b (alpha-IFN) in hepatitis C positive patients in the early posttransplant period and found that it appears to be a safe drug. There was no increase in rejection in patients receiving alpha-IFN, and patient and graft survival were the same as in our overall patient population. A combination a-IFN with Ribavirin will be undertaken in the near future. The use of Lamivudine in hepatitis B patients was shown to be effective in preventing and treating recurrence of hepatitis B posttransplant. Unmodified donor bone marrow cells (DBMC) were isolated from the vertebral bodies of the same cadaveric liver donors. Donor bone marrow dose, number of cells and/or number (or timing) of infusions were investigated to determine which variables affected the ability of DBMC to engraft in the liver recipient. The long-term benefit of DBMC needs further follow-up. Although, our patient and graft survival for liver transplant recipients is comparable to other large centers nationally and internationally, we still have some challenges to overcome. These include: 1) control and prevention of recurrent HCV, 2) improved treatment for hepatocellular cancer pre- and posttransplant, and 3) treatment and prevention of chronic rejection. Intestinal transplantation remains an even greater challenge. Diagnostic tests to determine intestinal function need further development and although MMF has shown some promise in this field, newer immunosuppressive medications need to be investigated to prevent rejection and avoid over immunosuppression.


[Technics for the morphologic study of leiomyoma of the bladder surface in women].
We present a case of leiomyoma of the vesical trigone in women, which was studied with intravenous urography, serial micturition cystourethrography, computerized axial tomography, abdominal echography and transvaginal echography. Comparing the data obtained by each of these morphological study techniques with those found in surgery, we were able to appreciate the high precision of those obtained by transvaginal echography. We think that this technique should be taken into account in the study of masses located on the bladder surface in women.


Gamma-Glutamyl Transferase (γ-GT) - an old dog with new tricks?
Gamma-Glutamyl Transferase (γGT) is a key transferase involved in the transpeptidation of functional gamma-glutamyl groups to various receptor moieties. It performs important roles in antioxidant defence mechanisms, particularly glutathione recycling, xenobiotic metabolism, but analogously may also have a pro-oxidant role. γGT is very sensitive for the diagnosis of liver injury, although it has poor specificity for particular aetiologies. It has been used to reflect temporal changes as a form of monitoring depending on aetiology. Given its cellular role in antioxidant function, it has been investigated as a surrogate biomarker of oxidative stress. It has also been found to be a predictor of mortality across a spectra of non-hepatic disease pathologies, from metabolic and cardiovascular risk to chronic kidney disease and neoplasia. Similarly, it also remains of interest to the insurance industry given an apparent ability to predict mortality, in addition to a historical interest from law enforcement as a marker of chronic alcohol ingestion. Here, we review some of the unique characteristics of this important enzyme, previously considered as a mere specific marker of liver dysfunction, but now with clear extra-hepatic implications and novel applications and utility.


Botulinum Toxin A and Lower Urinary Tract Dysfunction: Pathophysiology and Mechanisms of Action.
The use of onabotulinumtoxinA (BoNT-A) for the treatment of lower urinary tract diseases (LUTD) has increased markedly in recent years. The indications for BoNT-A treatment of LUTD now include neurogenic or idiopathic detrusor overactivity, interstitial cystitis/bladder pain syndrome and voiding dysfunction. The mechanisms of BoNT-A action on LUTDs affect many different aspects. Traditionally, the effects of BoNT-A were believed to be attributable to inhibition of acetylcholine release from the presynaptic efferent nerves at the neuromuscular junctions in the detrusor or urethral sphincter. BoNT-A injection in the bladder also regulated sensory nerve function by blocking neurotransmitter release and reducing receptor expression in the urothelium. In addition, recent studies revealed an anti-inflammatory effect for BoNT-A. Substance P and nerve growth factor in the urine and bladder tissue decreased after BoNT-A injection. Mast cell activation in the bladder also decreased. BoNT-A-induced improvement of urothelium function plays an important mitigating role in bladder dysfunction. Vascular endothelial growth factor expression in urothelium decreased after BoNT-A injection, as did apoptosis. Studies also revealed increased apoptosis in the prostate after BoNT-A injection. Although BoNT-A injection has been widely used to treat different LUTDs refractory to conventional treatment, currently, onabotulinumtoxinA has been proven effective only on urinary incontinence due to IDO and NDO in several large-scale clinical trials. The effects of onabotulinumtoxinA on other LUTDs such as interstitial cystitis, benign prostatic hyperplasia, dysfunctional voiding or detrusor sphincter dyssynergia have not been well demonstrated.


Prevention of chemotherapy-induced alopecia using an effective scalp cooling system.
Alopecia is a distressing side-effect of cancer treatment. Taxanes (TX), anthracyclines (ANR) and etoposide (ET) have been consistently associated with significant alopecia. We studied an effective scalp cooling system, the Penguin Cold Cap system, for the prevention of chemotherapy-induced alopecia in 70 patients receiving chemotherapy, including one of the following major alopecia-causing agents: Group A, TX-based regimes (without ANR); Group B, TX+ANR; Group C, ANR-based regimes (without TX); Group D, ET-based regimes. Protection from hair loss was achieved by maintaining scalp temperatures below 15 degrees C before, during and after chemotherapy by frequent changing of the caps. Assessment was carried out using a grading system from 0 to 4. Grades 0-2 were considered as satisfactory hair protection, whilst Grades 3-4 were considered failures. 57 patients were evaluable for assessment. An overall 81% protection was achieved. In groups C and D 11 of 12 patients (92%) had no alopecia, whilst 30 of 34 patients (88%) treated with taxanes had adequate hair protection. In Group B, 4 of 11 patients (36%) had adequate hair protection. The system was well tolerated and is a very effective method for protection from hair loss caused by TX, ANR and ET. Our results are comparable with and, in most cases, better than those reported in other studies using various alopecia preventive methods.


Molecular events in the induction of a nonresponsive state in interleukin 2-producing helper T-lymphocyte clones.
Exposure of normal interleukin 2 (IL-2)-producing helper T-cell clones to antigen and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-treated antigen-presenting cells results in proliferative unresponsiveness to subsequent stimulation with antigen and normal antigen-presenting cells. In the present study, we have examined the molecular events that accompany the induction of this unresponsive state. T cells stimulated in this manner failed to produce IL-2, but interleukin 3, interferon-gamma, and IL-2 receptors were partially induced and T-cell receptor beta mRNA was fully induced. Although T-cell unresponsiveness correlated with an IL-2 production defect, addition of IL-2 during the induction phase failed to prevent development of the unresponsive state. The critical biochemical event appeared to be an increase in intracellular calcium. Removal of calcium from the medium prevented induction of the unresponsive state, whereas addition of the calcium ionophore ionomycin induced unresponsiveness as well as all of the related partial activation events. Thus, an increase in intracellular calcium under nonmitogenic conditions appears to initiate an alternative activation program that prevents the T cell from producing IL-2 in response to subsequent normal activation signals. The significance of this in vitro model for tolerance induction in vivo is discussed.


Chapter 9: Asthma classification.
Asthma is a chronic inflammatory disorder of the airways resulting physiologically in hyperreactivity and clinically in recurrent episodes of wheezing, chest tightness, or coughing. Airway inflammation, smooth muscle contraction, epithelial sloughing, mucous hypersecretion, bronchial hyperresponsiveness, and mucosal edema contribute to the underlying pathophysiology of asthma. Diagnostic tests such as methacholine or mannitol challenges or spirometry (pre- and postbronchodilator responses) help to identify such underlying pathophysiology via assessments of bronchial hyperreactivity and lung mechanics but are imperfect and ultimately must be viewed in the context of a patient's clinical presentation including response to pharmacotherapy. The National Asthma Education and Prevention Program Expert Panel Report (2007) classifies asthma into either intermittent or persistent, and the latter is either mild, moderate, or severe. Some patients change in either direction from intermittent to persistent asthma. In addition, patients with asthma may be classified as allergic (IgE mediated), nonallergic (often triggered by viral upper respiratory tract infections or no apparent cause), occupational, aspirin-exacerbated respiratory disease, potentially (near) fatal, exercise induced, and cough variant asthma. In the latter, the patients have a nonproductive cough that responds to treatment for asthma but not with antibiotics, expectorants, mucolytics, antitussives, beta(2)-adrenergic agonists, treatment for acid reflux, or rhinosinusitis. Thus, cough variant asthma is in the differential diagnosis of chronic cough.


Detergent-dependent kinetics of truncated Plasmodium falciparum dihydroorotate dehydrogenase.
The survival of the malaria parasite Plasmodium falciparum is dependent upon the de novo biosynthesis of pyrimidines. P. falciparum dihydroorotate dehydrogenase (PfDHODH) catalyzes the fourth step in this pathway in an FMN-dependent reaction. The full-length enzyme is associated with the inner mitochondrial membrane, where ubiquinone (CoQ) serves as the terminal electron acceptor. The lipophilic nature of the co-substrate suggests that electron transfer to CoQ occurs at the two-dimensional lipid-solution interface. Here we show that PfDHODH associates with liposomes even in the absence of the N-terminal transmembrane-spanning domain. The association of a series of ubiquinone substrates with detergent micelles was studied by isothermal titration calorimetry, and the data reveal that CoQ analogs with long decyl (CoQ(D)) or geranyl (CoQ(2)) tails partition into detergent micelles, whereas that with a short prenyl tail (CoQ(1)) remains in solution. PfDHODH-catalyzed reduction of CoQ(D) and CoQ(2), but not CoQ(1), is stimulated as detergent concentrations (Tween 80 or Triton X-100) are increased up to their critical micelle concentrations, beyond which activity declines. Steady-state kinetic data acquired for the reaction with CoQ(D) and CoQ(2) in substrate-detergent mixed micelles fit well to a surface dilution kinetic model. In contrast, the data for CoQ(1) as a substrate were well described by solution steady-state kinetics. Our results suggest that the partitioning of lipophilic ubiquinone analogues into detergent micelles needs to be an important consideration in the kinetic analysis of enzymes that utilize these substrates.


Evaluation of Barley Cultivars for Competitive Traits in Southern New South Wales.
The potential of multi-purpose barley (Hordeum vulgare L.) cultivars to suppress weeds while maintaining optimal yield and grain quality has been reported but not recently evaluated in replicated field trials performed under southern Australian field conditions. Therefore, to investigate this potential, aboveground competitive traits were assessed in nine genetically diverse commercial barley cultivars in 2015, 2016 and 2017, in two locations in the Riverina region of NSW in replicated field trials performed in the absence of pre-emergent herbicide treatment. Crop and weed establishment, early vigour, leaf area index, photosynthetically active radiation (PAR) and biomass were assessed at various crop growth stages, including early growth, vegetative, flowering, grain fill and harvest. Cultivar differences in crop and weed biomass accumulation at ~50, 100 and 150 days after planting were noted at both locations. Early barley biomass accumulation was inversely related to weed biomass in both locations and most years, suggesting strong (over 90%) potential for heritable competitive barley interference against weeds. The current study also observed a positive relationship between PAR light interception and crop biomass in all three years at both locations, suggesting that PAR light interception contributed positively to crop biomass accumulation by directly increasing photosynthesis (50-70%) and growth or indirectly influencing weed biomass accumulation (10-15%) and weed interference (50-75). Partial least squares modelling was performed with 2015 and 2016 datasets to assess the interactions between crop developmental traits and weed suppression. Cultivars exhibiting enhanced early vigour and PAR light interception were generally more weed suppressive under optimal higher soil moisture conditions. Our results indicate that the choice of barley cultivar has a significant impact on weed establishment, fecundity and seedbank dynamics. The use of competitive barley genotypes is, thus, a cost-effective strategy to reduce weed seedbank numbers over time and may reduce potential herbicide use.


A selective 5-HT1a receptor agonist improves respiration in a mouse model of Rett syndrome.
Rett syndrome is a neurological disorder caused by loss of function mutations in the gene that encodes the DNA binding protein methyl-CpG-binding protein 2 (Mecp2). A prominent feature of the syndrome is disturbances in respiration characterized by frequent apnea and an irregular interbreath cycle. 8-Hydroxy-2-dipropylaminotetralin has been shown to positively modulate these disturbances (Abdala AP, Dutschmann M, Bissonnette JM, Paton JF, Proc Natl Acad Sci U S A 107: 18208-18213, 2010), but the mode of action is not understood. Here we show that the selective 5-HT1a biased agonist 3-chloro-4-fluorophenyl-(4-fluoro-4-{[(5-methylpyrimidin-2-ylmethyl)-amino]-methyl}-piperidin-1-yl)-methanone (F15599) decreases apnea and corrects irregularity in both heterozygous Mecp2-deficient female and in Mecp2 null male mice. In whole cell voltage-clamp recordings from dorsal raphe neurons, F15599 potently induced an outward current, which was blocked by barium, reversed at the potassium equilibrium potential, and was antagonized by the 5-HT1a antagonist WAY100135. This is consistent with somatodendritic 5-HT1a receptor-mediated activation of G protein-coupled inwardly rectifying potassium channels (GIRK). In contrast, F15599 did not activate 5-HT1b/d receptors that mediate inhibition of glutamate release from terminals in the nucleus accumbens by a presynaptic mechanism. Thus F15599 activated somatodendritic 5-HT1a autoreceptors, but not axonal 5-HT1b/d receptors. In unanesthetized Mecp2-deficient heterozygous female mice, F15599 reduced apnea in a dose-dependent manner with maximal effect of 74.5 ± 6.9% at 0.1 mg/kg and improved breath irrregularity. Similarly, in Mecp2 null male mice, apnea was reduced by 62 ± 6.6% at 0.25 mg/kg, and breathing became regular. The results indicate respiration is improved with a 5-HT1a agonist that activates GIRK channels without affecting neurotransmitter release.


Dietary quality among men and women in 187 countries in 1990 and 2010: a systematic assessment.
Healthy dietary patterns are a global priority to reduce non-communicable diseases. Yet neither worldwide patterns of diets nor their trends with time are well established. We aimed to characterise global changes (or trends) in dietary patterns nationally and regionally and to assess heterogeneity by age, sex, national income, and type of dietary pattern. In this systematic assessment, we evaluated global consumption of key dietary items (foods and nutrients) by region, nation, age, and sex in 1990 and 2010. Consumption data were evaluated from 325 surveys (71·7% nationally representative) covering 88·7% of the global adult population. Two types of dietary pattern were assessed: one reflecting greater consumption of ten healthy dietary items and the other based on lesser consumption of seven unhealthy dietary items. The mean intakes of each dietary factor were divided into quintiles, and each quintile was assigned an ordinal score, with higher scores being equivalent to healthier diets (range 0-100). The dietary patterns were assessed by hierarchical linear regression including country, age, sex, national income, and time as exploratory variables. From 1990 to 2010, diets based on healthy items improved globally (by 2·2 points, 95% uncertainty interval (UI) 0·9 to 3·5), whereas diets based on unhealthy items worsened (-2·5, -3·3 to -1·7). In 2010, the global mean scores were 44·0 (SD 10·5) for the healthy pattern and 52·1 (18·6) for the unhealthy pattern, with weak intercorrelation (r=-0·08) between countries. On average, better diets were seen in older adults compared with younger adults, and in women compared with men (p<0·0001 each). Compared with low-income nations, high-income nations had better diets based on healthy items (+2·5 points, 95% UI 0·3 to 4·1), but substantially poorer diets based on unhealthy items (-33·0, -37·8 to -28·3). Diets and their trends were very heterogeneous across the world regions. For example, both types of dietary patterns improved in high-income countries, but worsened in some low-income countries in Africa and Asia. Middle-income countries showed the largest improvement in dietary patterns based on healthy items, but the largest deterioration in dietary patterns based on unhealthy items. Consumption of healthy items improved, while consumption of unhealthy items worsened across the world, with heterogeneity across regions and countries. These global data provide the best estimates to date of nutrition transitions across the world and inform policies and priorities for reducing the health and economic burdens of poor diet quality. The Bill & Melinda Gates Foundation and Medical Research Council.


Lymphoepithelial carcinoma in the sublingual gland.
Lymphoepithelial carcinoma is rare in the salivary glands, with an incidence of 0.4%. The most commonly affected site is the parotid gland, followed by the submandibular gland. Lymphoepithelial carcinoma in the sublingual gland has been reported only four times in the existing English-language literature. Such tumours are characterized by the presence of a poorly differentiated carcinoma that is surrounded and infiltrated by lymphocytes, and they are strongly associated with Epstein-Barr virus infection, patient ethnicity, and prominent radiosensitivity. Wide surgical excision combined with adjuvant therapy has been suggested as the first-choice therapeutic regimen. This report describes the case of a 34-year-old Indonesian woman who was evaluated and treated in Taipei Medical University Hospital. She had a tumour that presented as a painless swelling on the floor of the mouth. The diagnosis was confirmed by conducting an incisional biopsy, and a wide surgical excision with bilateral supraomohyoid neck dissection and free flap reconstruction was performed. The patient also underwent adjuvant chemoradiotherapy. No evidence of local recurrence or distant metastasis was detected during the 6 months of follow-up. Subsequently, the patient returned to her home country, and further follow-ups were not conducted.


Effect of weaning in the pig on ileal ion transport measured in vitro.
Early weaning in pigs results in small intestinal malabsorption and an increased susceptibility to E. coli infections. This is closely associated with villus shortening and crypt hyperplasia of the small intestine. The present study compared piglets either weaned at 3 weeks of age onto a high soya diet (n = 12) or an egg-based diet (n = 12) with piglets that remained on the sow (n = 12). Prior to weaning, care was taken to ensure that piglets only had access to sows milk. Serum anti-soya IgG was measured 7 days after weaning and sections of the mid-ileum excised and fixed for determination of crypt depth and villus height. Four pieces of 'stripped' mucosa were mounted in Ussing chambers in Krebs-phosphate Ringer (with indomethacin) for determination of short circuit current (SCC) and unidirectional fluxes of Na22 and Cl36, half in mucosa-serosa (MS), and half in serosa-mucosa (SM) direction. After basal measurements of absorptive capacity of Na, supramaximal doses of prostaglandin E2 (PGE2) and theophylline were added to measure the intestinal secretory capacity for Cl. Anti-soya IgG was elevated in the group weaned onto soya when compared either with the group weaned onto egg or the unweaned group. All intestinal transport and histological parameters were similar in both the weaned groups, although some were different from the unweaned. The SCC (equal to Na absorption) and the villus height were reduced two-thirds by weaning, whereas crypt depth and Cl secretion were similar in all groups. It is suggested that weaning per se is causal for the observed changes in intestinal transport and morphology, but they are not influenced by whether the weaning diet is soya or egg based.


The Key Glycolytic Enzyme Phosphofructokinase Is Involved in Resistance to Antiplasmodial Glycosides.
Plasmodium parasites rely heavily on glycolysis for ATP production and for precursors for essential anabolic pathways, such as the methylerythritol phosphate (MEP) pathway. Here, we show that mutations in the Plasmodium falciparum glycolytic enzyme, phosphofructokinase (PfPFK9), are associated with in vitro resistance to a primary sulfonamide glycoside (PS-3). Flux through the upper glycolysis pathway was significantly reduced in PS-3-resistant parasites, which was associated with reduced ATP levels but increased flux into the pentose phosphate pathway. PS-3 may directly or indirectly target enzymes in these pathways, as PS-3-treated parasites had elevated levels of glycolytic and tricarboxylic acid (TCA) cycle intermediates. PS-3 resistance also led to reduced MEP pathway intermediates, and PS-3-resistant parasites were hypersensitive to the MEP pathway inhibitor, fosmidomycin. Overall, this study suggests that PS-3 disrupts core pathways in central carbon metabolism, which is compensated for by mutations in PfPFK9, highlighting a novel metabolic drug resistance mechanism in P. falciparumIMPORTANCE Malaria, caused by Plasmodium parasites, continues to be a devastating global health issue, causing 405,000 deaths and 228 million cases in 2018. Understanding key metabolic processes in malaria parasites is critical to the development of new drugs to combat this major infectious disease. The Plasmodium glycolytic pathway is essential to the malaria parasite, providing energy for growth and replication and supplying important biomolecules for other essential Plasmodium anabolic pathways. Despite this overreliance on glycolysis, no current drugs target glycolysis, and there is a paucity of information on critical glycolysis targets. Our work addresses this unmet need, providing new mechanistic insights into this key pathway.


Assessing the genetic diversity in Argopecten nucleus (Bivalvia: Pectinidae), a functional hermaphrodite species with extremely low population density and self-fertilization: Effect of null alleles.
Argopecten nucleus is a functional hermaphroditic pectinid species that exhibits self-fertilization, whose natural populations have usually very low densities. In the present study, the genetic diversity of a wild population from Neguanje Bay, Santa Marta (Colombia), was estimated using microsatellite markers, and the effect of the presence of null alleles on this estimation was assessed. A total of 8 microsatellite markers were developed, the first described for this species, and their amplification conditions were standardized. They were used to determine the genotype of 48 wild individuals from Naguanje Bay, and 1,010 individuals derived from the offspring of 38 directed crosses. For each locus, the frequencies of the identified alleles, including null alleles, were estimated using the statistical package Micro-Checker, and the parental genotypes were confirmed using segregation analysis. Three to 8 alleles per locus with frequencies from 0.001 to 0.632 were detected. The frequencies of null alleles ranged from 0.10 to 0.45, with Ho from 0.0 to 0.79, and He from 0.53 to 0.80. All loci were in H-W disequilibrium. The null allele frequencies values were high, with lower estimations using segregation analysis than estimated using Micro-Checker. The present results show high levels of population genetic diversity and indicate that null alleles were not the only cause of deviation from H-W equilibrium in all loci, suggesting that the wild population under study presents signs of inbreeding and Wahlund effect.


Tomato transcriptional changes in response to Clavibacter michiganensis subsp. michiganensis reveal a role for ethylene in disease development.
Clavibacter michiganensis subsp. michiganensis (Cmm) is a gram-positive actinomycete, causing bacterial wilt and canker disease in tomato (Solanum lycopersicum). Host responses to gram-positive bacteria and molecular mechanisms associated with the development of disease symptoms caused by Cmm in tomato are largely unexplored. To investigate plant responses activated during this compatible interaction, we used microarray analysis to monitor changes in host gene expression during disease development. This analysis was performed at 4 d postinoculation, when bacteria were actively multiplying and no wilt symptoms were yet visible; and at 8 d postinoculation, when bacterial growth approached saturation and typical wilt symptoms were observed. Of the 9,254 tomato genes represented on the array, 122 were differentially expressed in Cmm-infected plants, compared with mock-inoculated plants. Functional classification of Cmm-responsive genes revealed that Cmm activated typical basal defense responses in the host, including induction of defense-related genes, production and scavenging of free oxygen radicals, enhanced protein turnover, and hormone synthesis. Cmm infection also induced a subset of host genes involved in ethylene biosynthesis and response. After inoculation with Cmm, Never ripe (Nr) mutant plants, impaired in ethylene perception, and transgenic plants with reduced ethylene synthesis showed significant delay in the appearance of wilt symptoms, compared with wild-type plants. The retarded wilting in Nr plants was a specific effect of ethylene insensitivity, and was not due to altered expression of defense-related genes, reduced bacterial populations, or decreased ethylene synthesis. Taken together, our results indicate that host-derived ethylene plays an important role in regulation of the tomato susceptible response to Cmm.


Expression Profiling of MicroRNAs in the Inner Ear of Elderly People by Real-Time PCR Quantification.
The molecular mechanisms underlying age-related hearing loss are unknown, and currently, there is no treatment for this condition. Recent studies have shown that microRNAs (miRNAs) and age-related diseases are intimately linked, suggesting that some miRNAs may present attractive therapeutic targets. In this study, we obtained 8 human temporal bones from 8 elderly subjects at brain autopsy in order to investigate the expression profile of miRNAs in the inner ear with miRNA arrays. A mean of 478 different miRNAs were expressed in the samples, of which 348 were commonly expressed in all 8 samples. Of these, levels of 16 miRNAs significantly differed between young elderly and old elderly subjects. miRNAs, which play important roles in inner ear development, were detected in all samples, i.e., in both young and old elderly subjects, whether with or without hearing loss. Our results suggest that these miRNAs play important roles not only in development, but also in the maintenance of inner ear homeostasis.


Suppressed Size Effect in Nanopillars with Hierarchical Microstructures Enabled by Nanoscale Additive Manufacturing.
Studies on mechanical size effects in nanosized metals unanimously highlight both intrinsic microstructures and extrinsic dimensions for understanding size-dependent properties, commonly focusing on strengths of uniform microstructures, e.g., single-crystalline/nanocrystalline and nanoporous, as a function of pillar diameters, D. We developed a hydrogel infusion-based additive manufacturing (AM) technique using two-photon lithography to produce metals in prescribed 3D-shapes with ∼100 nm feature resolution. We demonstrate hierarchical microstructures of as-AM-fabricated Ni nanopillars (D ∼ 130-330 nm) to be nanoporous and nanocrystalline, with d ∼ 30-50 nm nanograins subtending each ligament in bamboo-like arrangements and pores with critical dimensions comparable to d. In situ nanocompression experiments unveil their yield strengths, σ, to be ∼1-3 GPa, above single-crystalline/nanocrystalline counterparts in the D range, a weak size dependence, σ ∝ D-0.2, and localized-to-homogenized transition in deformation modes mediated by nanoporosity, uncovered by molecular dynamics simulations. This work highlights hierarchical microstructures on mechanical response in nanosized metals and suggests small-scale engineering opportunities through AM-enabled microstructures.


The development and goals of the AAFP center for policy studies in family practice and primary care. American Academy of Family Physicians.
In this article we describe the creation and role of the Center for Policy Studies in Family Practice and Primary Care established by the American Academy of Family Physicians in Washington, DC, this year. We recount the events leading to the decision to implement the Center, list its guiding assumptions, and explain its initial structure and function. We also identify the 3 themes that will guide the early work of the Center: sustaining the functional domain of family practice and primary care; investing in key infrastructures; and securing universal health coverage.


Effect of garlic supplementation to atherogenic diet on collagen biosynthesis in various tissues of rabbits.
Feeding of cholesterol-rich diet to male rabbits increases collagen biosynthesis and accumulation in aorta, liver, kidney, heart and lungs. However, when garlic supplemented atherogenic diet was fed to the animals, the above effects were partially suppressed particularly in the aorta. Cholesterol content of plasma, aorta and liver, which was tremendously increased on atherogenic diet, was suppressed when garlic was included in this diet. It is suggested that garlic possibly reduced the accumulation of collagen through more mobilisation of lipids and/or decreased biosynthesis and maturation of collagen. Histopathological studies also supported this view.


Contrast lipocryolysis: Pre- and post-session tempering improves clinical results.
Alternative crystal structures are possible for all lipids and each different crystal structure is called a polymorphic form. Inter-conversion between polymorphisms would imply the possibility of leaning crystal formation toward the most effective polymorphism for adipocyte destruction. Food industry has been tempering lipids for decades. Tempering technology applied to lipocryolysis gave birth to "contrast lipocryolysis", which involves pre- and post-lipocryolysis fat layer heating as part of a specific tempering protocol. In this study, we evaluated the skinfold thickness of 10 subjects after a single contrast lipocryolysis session and witnessed important and fast reductions.


Emotional, motivational and interpersonal responsiveness of children with autism in improvisational music therapy.
Through behavioural analysis, this study investigated the social-motivational aspects of musical interaction between the child and the therapist in improvisational music therapy by measuring emotional, motivational and interpersonal responsiveness in children with autism during joint engagement episodes. The randomized controlled study (n = 10) employed a single subject comparison design in two different conditions, improvisational music therapy and toy play sessions, and DVD analysis of sessions. Improvisational music therapy produced markedly more and longer events of 'joy', 'emotional synchronicity' and 'initiation of engagement' behaviours in the children than toy play sessions. In response to the therapist's interpersonal demands, 'compliant (positive) responses' were observed more in music therapy than in toy play sessions, and 'no responses' were twice as frequent in toy play sessions as in music therapy. The results of this exploratory study found significant evidence supporting the value of music therapy in promoting social, emotional and motivational development in children with autism.


Ultrastructural studies on the neuromuscular control of human tracheal and bronchial muscle.
This study presents an ultrastructural analysis of neural and myogenic control of smooth muscle in human trachea, in small (approximately fourth to seventh order) bronchi. A moderate frequency of gap junctions between smooth muscle cells and a sparse innervation was observed in trachea. In contrast, small junctions without identifiable gaps were identified in bronchi along with a dense, sometimes close innervation of muscle. Most profiles of nerve varicosities in both types of airways were of two types: (1) those containing mostly small agranular and few or no large granular vesicles; and (2) nerves (probably adrenergic) with some small granular vesicles along with other vesicles present in bronchi. In the bronchi some mast cells were found near nerve profiles and in close proximity to smooth muscle. These morphological findings suggest that tracheal muscle may be organized for more neural and less myogenic control of activity. Interactions between mast cells, nerves and muscles in bronchi could influence airway function. These findings imply that a single type of airway muscle may not adequately represent the structural and functional properties of human airway.


Urothelial carcinoma of the renal pelvis: a clinicopathologic study of 130 cases.
Urothelial carcinomas of the renal pelvis are relatively rare tumors, and large series that include clinicopathologic and outcome data are few. We reviewed 130 consecutive nephroureterectomies performed for urothelial carcinoma of the renal pelvis at our institution. Tumors were graded using the World Health Organization/International Society of Urologic Pathologists (WHO/ISUP) grading system and were staged according to the 2002 TNM classification; 83 (63.6%) of the patients were men and 47 (36.4%) were women. The mean age at diagnosis was 67 years (range, 41-93 years). The average tumor size was 3.7 cm; 36 of the cases were multifocal and 5 were bilateral. Lower tract disease occurred in 50.7% (66 cases); 38 of the cases (29.3%) were low grade and the remaining 92 (70.7%) were high grade. A total of 50% of the cases were pTis, pTa, or pT1, while 45% invaded deeply (pT2 or more). Depth of invasion could not be assessed with certainty in 7 cases (5%). Regional lymph nodes were identified/submitted in only 50 cases. Of those, 12 cases (24%) had lymph node metastasis. Follow-up information was available in 125 (96%) patients. The period of follow-up ranged from 1 week to 176 months (mean, 48.9 months). At last follow-up, 47 patients (36%) had died of other causes, 18 (13.8%) were dead of disease, 8 patients (6%) were alive with disease, and 52 patients (40%) were alive with no evidence of disease. In univariate analysis, histologic grade (P = 0.001), TNM stage (P = 0.0001), vascular invasion (P = 0.001), margin status (P = 0.021), and size (P = 0.0003) were significantly associated with survival. On multivariate analysis, TNM stage (P = 0.03) was the only variable associated with survival. In conclusion, our study shows that a high percentage of the urothelial carcinoma of the renal pelvis present with locally advanced (pT2 or more) disease at the time of nephroureterectomy. Pathologic stage is the most potent predictor of survival, similar to lower tract disease. A subset of the cases could not be staged due to processing issues; we thus recommend fixation prior to prosecting.


Microsurgery in urology.
Microsurgery is becoming increasingly widely accepted in the field of urology, and microsurgical training well become an integral part of the programs for Residents in University Schools of Urology. In this paper we outline the present clinical application of microsurgery in urology and perspectives, for example: workbench surgery, testis transplantation, vasovasostomy, revascularization for impotence, penis replantation, microsurgery of the urinary tract.


Protein misfolding and the pathogenesis of ABCA4-associated retinal degenerations.
Mutations in the ABCA4 gene are a common cause of autosomal recessive retinal degeneration. All mouse models to date are based on knockouts of Abca4, even though the disease is often caused by missense mutations such as the complex allele L541P;A1038V (PV). We now show that the PV mutation causes severe human disease whereas the V mutation alone causes mild disease. Mutant ABCA4 proteins expressed heterologously in mammalian cells retained normal cellular localization. However, basal and all-trans-retinal-stimulated ATPase activities were reduced substantially for P and PV but only mildly for V. Electron microscopy revealed marked structural changes and misfolding for the P and PV mutants but few changes for the V mutant, consistent with the disease severity difference in patients. We generated Abca4(PV/PV) knock-in mice homozygous for the complex PV allele to investigate the effects of this misfolding mutation in vivo. Mutant ABCA4 RNA levels approximated WT ABCA4 RNA levels but, surprisingly, only trace amounts of mutant ABCA4 protein were noted in the retina. RNA sequencing of WT, Abca4(-/-) and Abca4(PV/PV) mice revealed mild gene expression alterations in the retina and RPE. Similar to Abca4(-/-) mice, Abca4(PV/PV) mice showed substantial A2E and lipofuscin accumulation in their RPE cells but no retinal degeneration up to 12 months of age. Thus, rapid degradation of this large misfolded mutant protein in mouse retina caused little detectable photoreceptor degeneration. These findings suggest likely differences in the unfolded protein response between murine and human photoreceptors and support development of therapies directed at increasing this capability in patients.


Diversity and phylogenetic relationships among Bartonella strains from Thai bats.
Bartonellae are phylogenetically diverse, intracellular bacteria commonly found in mammals. Previous studies have demonstrated that bats have a high prevalence and diversity of Bartonella infections globally. Isolates (n = 42) were obtained from five bat species in four provinces of Thailand and analyzed using sequences of the citrate synthase gene (gltA). Sequences clustered into seven distinct genogroups; four of these genogroups displayed similarity with Bartonella spp. sequences from other bats in Southeast Asia, Africa, and Eastern Europe. Thirty of the isolates representing these seven genogroups were further characterized by sequencing four additional loci (ftsZ, nuoG, rpoB, and ITS) to clarify their evolutionary relationships with other Bartonella species and to assess patterns of diversity among strains. Among the seven genogroups, there were differences in the number of sequence variants, ranging from 1-5, and the amount of nucleotide divergence, ranging from 0.035-3.9%. Overall, these seven genogroups meet the criteria for distinction as novel Bartonella species, with sequence divergence among genogroups ranging from 6.4-15.8%. Evidence of intra- and intercontinental phylogenetic relationships and instances of homologous recombination among Bartonella genogroups in related bat species were found in Thai bats.


Development of cerebral gray and white matter injury and cerebral inflammation over time after inflammatory perinatal asphyxia.
Antenatal inflammation is associated with increased severity of hypoxic-ischemic (HI) encephalopathy and adverse outcome in human neonates and experimental rodents. We investigated the effect of lipopolysaccharide (LPS) on the timing of HI-induced cerebral tissue loss and gray matter injury, white matter injury and integrity, and the cerebral inflammatory response. On postnatal day 9, mice underwent HI by unilateral carotid artery occlusion followed by systemic hypoxia which resulted in early neuronal damage (MAP2 loss) at 3 h that did not increase up to day 15. LPS injection 14 h before HI (LPS+HI) significantly and gradually aggravated MAP2 loss from 3 h up to day 15, resulting in an acellular cystic lesion. LPS+HI increased white matter damage, reduced myelination in the corpus callosum and increased white matter fiber coherency in the cingulum. The number of oligodendrocytes throughout the lineage (Olig2-positive) was increased whereas more mature myelinating (CNPase-positive) oligodendrocytes were strongly decreased after LPS+HI. LPS+HI induced an increased and prolonged expression of cerebral cytokines/chemokines compared to HI. Additionally, LPS+HI increased macrophage/microglia activation and influx of neutrophils in the brain compared to HI. This study demonstrates the sensitizing effect of LPS on neonatal HI brain injury for an extended time-frame up to 15 days postinsult. LPS before HI induced a gradual increase in gray and white matter deficits, including reduced numbers of more mature myelinating oligodendrocytes and a decrease in white matter integrity. Moreover, LPS+HI prolonged and intensified the cerebral inflammatory response, including cellular infiltration. In conclusion, as the timing of damage and/or involved pathways are changed when HI is preceded by inflammation, experimental therapies might require modifications in the time window, dosage or combinations of therapies for efficacious neuroprotection.


Mus81-Eme1 and Rqh1 involvement in processing stalled and collapsed replication forks.
The processing of stalled replication forks and the repair of collapsed replication forks are essential functions in all organisms. In fission yeast DNA junctions at stalled replication forks appear to be processed by either the Rqh1 DNA helicase or Mus81-Eme1 endonuclease. Accordingly, we show that the hypersensitivity to agents that cause replication fork stalling of mus81, eme1, and rqh1 mutants is suppressed by a Holliday junction resolvase (RusA), as is the synthetic lethality of a mus81(-) rqh1(-) double mutant. Recombinant Mus81-Eme1, purified from Escherichia coli, readily cleaves replication fork structures but cleaves synthetic Holliday junctions relatively poorly in vitro. From these data we propose that Mus81-Eme1 can process stalled replication forks before they have regressed to form a Holliday junction. We also implicate Mus81-Eme1 and Rqh1 in the repair of collapsed replication forks. Here Mus81-Eme1 and Rqh1 seem to function on different substrates because RusA can substitute for Mus81-Eme1 but not Rqh1.


Benign prostatic hyperplasia (BPH) and prostatitis: prevalence of painful ejaculation in men with clinical BPH.
To determine the prevalence and importance of pain/discomfort on ejaculation (prostatitis-like symptom) in men with lower urinary tract symptoms (LUTS) diagnosed with clinical benign prostatic hyperplasia (BPH). Baseline data from 5096 men reporting LUTS suggestive of BPH, and enrolled in the ALF-ONE study by general practitioners and urologists in Europe, Asia, Latin America, the Middle East and Canada, were analysed to determine the prevalence and significance of pain/discomfort on ejaculation. All the men were asked to complete the International Prostate Symptom Score (IPSS) questionnaire, the bother score (IPSS question 8), and the Danish Prostate Symptom Score sexual-function questionnaire (DAN-PSSsex) which assesses three symptoms (rigidity of erection, amount of ejaculate and pain/discomfort on ejaculation) and their bothersomeness. There were 3700 sexually active men who had an evaluable answer to the DAN-PSSsex question related to pain/discomfort on ejaculation. Of these, 688 (18.6%) reported pain/discomfort on ejaculation and 609 (88%) considered it was a problem. Patients with painful ejaculation had more severe LUTS and reported greater bother (P < 0.001). Of men with painful ejaculation, 72% reported erectile dysfunction, of whom 91% considered it a problem, and 75% reported reduced ejaculation, of whom 81% considered it a problem. By contrast, of men with no ejaculatory discomfort, 57% reported erectile dysfunction, of whom 79% considered it a problem, and 56% reported reduced ejaculation, of whom 57% considered it a problem. A history of urinary tract infection was reported by 12% of men in the ejaculatory pain group, compared with 7% in the LUTS-only group, while 5% of men in the ejaculatory pain group reported macroscopic haematuria, compared to 3% in the LUTS-only group. Men with ejaculatory pain were slightly younger, but there were no significant differences in duration of LUTS, history of acute urinary retention, prostate-specific antigen concentrations or maximum urinary flow rate compared to the LUTS-only group. Of sexually active men with LUTS suggestive of BPH, approximately 20% complain of specific prostatitis-like symptoms of pain/discomfort on ejaculation, and these men clearly differ from those who present with LUTS only. For most the symptom is a significant bother. Men with BPH and painful ejaculation have more severe LUTS and reported greater bother, and had a higher prevalence of erectile dysfunction and reduced ejaculation, than men with LUTS only. Evaluation and treatment strategies should address this population of men with symptoms suggestive of both prostatitis and BPH.


[Burnout among Family Healthcare physicians: the challenge of transformation in the workplace].
The scope of this paper is to understand how conflicts with the institution and disagreements regarding team members'attributions are interpreted by Family Healthcare physicians from the burnout perspective. Qualitative research was conducted with 24 doctors in Recife between August 2005 and November 2006 based on management evaluation (geographic access, conflicts within the team, between teams and the District, between the team and the community and public violence in the area), and four teams were selected in each of the six Health Districts. Semi-structured interviews were then conducted. An enormous discrepancy between the expectations of physicians and the reality of the workplace was detected. They reject the institutional consultation priority and demands for productivity. They suffer from what they consider loss of professional identity, with nurses attributions questioning the historic essence of their practice. There is discredit concerning change and a desire to abandon the profession, especially amongst those with the highest expectations concerning conversion of the model. A broad set of elements was detected that favor simultaneous exhaustion and professional inefficiency and provoke negative attitudes, stressing the need for promoting health in the workplace.


Parental monitoring and youth behavior problems: moderation by callous-unemotional traits over time.
Prior research suggests that parents' monitoring behaviors are related to the conduct problems of children but not to the conduct problems of children with callous-unemotional traits. However, these studies have been cross-sectional. The present short-term longitudinal study investigates the bidirectional influences of parental monitoring and youth problem behaviors, while also examining the potential moderating influence of callous-unemotional traits. We assessed adolescents (Mean age = 14.5 years; SD = 1.8) and parents at two time points. Youths reported their callous-unemotional traits and delinquency, while parents reported their child's conduct problems, and their parenting behaviors to track and control their child's activities. We found support for a child-driven change in parents' monitoring behaviors over time. Specifically, children with high callous-unemotional traits had parents who reduced their monitoring behaviors over time, and their different types of monitoring behaviors were less synchronous over time. In addition, parents of youths with high callous-unemotional traits showed a trend toward not being stable in their surveillance efforts over time. Moreover, greater behavioral control for youths high on callous-unemotional traits did not lead to parents' greater knowledge about their youths. In fact, having less knowledge was related to decreases in parental control, when youths were high on callous-unemotional traits. The present study supports the importance of personality in shaping how parents actively monitor their children.


Hydroxysafflor yellow A increases BDNF and NMDARs in the hippocampus in a vascular dementia rat model.
Hydroxysafflor yellow A (HSYA) is a drug that exerts angiogenesis regulatory and neuroprotective effects and has become an effective therapy for brain and heart ischemic disorders. There is no definite evidence supporting a therapeutic effect of HSYA in vascular dementia (VaD). We used HSYA in a rat model of chronic cerebral ischemia to determine its potential therapeutic effects in VaD. The Morris water maze (MWM) was used to evaluate spatial cognitive function, and long-term potentiation (LTP) was tested as a marker of synaptic plasticity. The expression levels of brain-derived neurotrophic factor (BDNF) and two subunits of N-methyl-d-aspartate receptor (NMDAR; GluN2A and GluN2B) in the hippocampus were measured via western blotting. The MWM results showed that the experimental VaD group had longer escape latencies than the sham group, whereas the HSYA group had a decreased escape latency compared with the VaD group (P<0.05). The LTP at CA3-CA1 synapses in the hippocampus was also enhanced in the HSYA compared with the VaD group (P<0.05). The western blotting results revealed lower hippocampal BDNF and GluN2B expression in the VaD group compared with the sham group and significantly higher hippocampal expression in the HSYA group compared with the VaD group. No significant change in GluN2A expression was detected. The results indicate that HSYA may enhance the endogenous expression of BDNF and GluN2B, which are associated with the synaptic plasticity of the hippocampus, and may improve spatial learning and memory abilities in a rat model of VaD.


Efficacy and toxicity of drugs targeting KRASG12C mutation in non-small cell lung cancer: a meta-analysis.
To systematically analyze the efficacy and toxicity of drugs targeting KRASG12C mutation in non-small cell lung cancer (NSCLC). The candidate studies were identified in PubMed, Embase, Cochrane Library, CNKI, and Wanfang databases up to 1 June 2023. Data on efficacy, prognosis, and adverse events (AEs) were extracted and calculated by meta-analysis. Six eligible prospective studies were included in this meta-analysis, including 563 patients with advanced or metastatic NSCLC. For patients with NSCLC, the objective response rate (ORR) of drugs targeting KRASG12C mutation was 37% (95%CI 31-43), median duration of response (DOR) was 8.89 months (95%CI 7.96-9.83), and median progression-free survival (PFS) was 6.40 months (95%CI 5.86-6.93). The overall incidence of AEs was 88% (95%CI 79-96) and the incidence of grade ≥3 AEs was 44% (95%CI 24-64). The most common AEs were diarrhea, nausea, fatigue, and vomiting. The most common grade ≥3 AEs were Alaninetransaminase (ALT) or Aspartatetransaminase (AST) increased and diarrhea. Sotorasib, Adagrasib, and Garsorasib as the drugs of choice for patients with KRASG12C mutation NSCLC, have definite efficacy and acceptable safety, especially for patients with advanced or metastatic disease and within posterior line therapy.


Electrolyte transport in a central core model of the renal medulla.
Transport of Na+, K+, Cl-, urea, and water is described in a central core model of the renal medulla. Equations for mass balance, Poiseuille flow, and the Nernst-Planck equation describe the continuous behavior of the system along the medullary axis and along the distal nephron; the Kedem and Katchalsky phenomenology describes passive transmural transport; active transmural transport obeys Michaelis-Menten kinetics. Numerical solution of the differential equations shows that to a close approximation any combination of active Na+ and active Cl- transport can generate the same concentration profiles but will generate very different potential profiles, and consequently, very different K+ absorption in thick ascending limb of Henle's loop. If a net transport stoichiometry of 2 Cl- ions to 1 Na+ ion is selected for the pumps, an active Cl- transport rate of approximately 10,000 peq.s-1.cm-2 gives K+ and Na+ concentrations in early distal nephron and a medullary osmolality profile in reasonable agreement with experimental data.


[Establishing Evidence for Use of Appropriate Medicines in the Operating Room].
Various issues related to clinical use of medicines remain unclear, and pharmacists are expected to establish evidence for appropriate use of medicines. The present review summarizes our findings from three areas of research regarding the use of medicines in the operating room: 1) We evaluated the extent of extravasation injury due to thiopental (2.5 mg/100 μL) and propofol (1.0 mg/100 μL) at the macroscopic and histopathologic levels in a rat model. Thiopental, which causes tissue necrosis, can be classified as a "vesicant", and propofol can be classified as an "irritant". Moreover, warming strongly exacerbated the degeneration or necrosis induced by extravasation of thiopental. 2) The cytotoxicity of povidone-iodine solution (PVP-I) for ophthalmic use and that of polyvinyl alcohol-iodine solution (PAI) was compared using a human corneal epithelial cell line. Despite exhibiting equivalent antiseptic effects, the cytotoxicity of PVP-I diluted 16-fold was greater than that of PAI diluted 6-fold. After inactivation of iodine, the cytotoxicity of PVP-I persisted; therefore, to avoid corneal damage, antisepsis should be achieved with PAI. 3) The stability of 1 μg/mL adrenaline when used as an intraocular irrigating solution to maintain pupil dilation was evaluated. After mixing for 6 h, the adrenaline concentration was 65.2% (pH 8.0) of the initial concentration. Moreover, the low concentration of sodium bisulfite in the irrigating solution could have caused adrenaline reduction. Our results strongly suggest that intraocular irrigation solution containing adrenaline should be prepared just prior to use in surgery.


Respiratory pattern changes in sleep in children on vagal nerve stimulation for refractory epilepsy.
An altered breathing pattern in sleep, over two to three weeks, reported by the parents of a child on Vagal Nerve Stimulation (VNS) therapy for refractory epilepsy, prompted a sleep study in him. His polysomnography (PSG) revealed respiratory irregularity concordant with VNS activation. Dyspnoea is a well recognised and reported side effect of the VNS. However there are only a few studies looking at respiration in sleep with VNS. We therefore undertook PSGs in seven other children on VNS. Sleep studies were undertaken, in accordance with standard clinical practice. Sleep and apnoeas and hypopneas were scored in accordance with conventional criteria. Respiratory pattern changes in sleep (RPCS) with VNS were looked for. Respiratory pattern changes in sleep were seen during PSG in seven of eight children on VNS for refractory epilepsy. Decreased effort and tidal volume occurred in seven children, concordant with VNS activation. In one child, this was associated with a fall in respiratory rate, i the other six children with an increase. No study showed an apnoea/hypopnoea index in the abnormal range. The RPCS were not associated with significant hypoxia or hypercapnoea. Our results suggest that RPCS occur in most children with VNS. This is not surprising in view of the significant influence vagal afferents have on respiratory control centres. The RPCS did not appear to have a clinical impact in our group. However further investigations are suggested to explore this phenomenon, especially in patients with sleep apnoea syndromes or compromised respiratory function.


In vivo Mechanisms of Antibody-Mediated Neurological Disorders: Animal Models and Potential Implications.
Over the last two decades, the discovery of antibodies directed against neuronal surface antigens (NSA-Abs) in patients with different forms of encephalitis has provided a basis for immunotherapies in previously undefined disorders. Nevertheless, despite the circumstantial clinical evidence of the pathogenic role of these antibodies in classical autoimmune encephalitis, specific criteria need to be applied in order to establish the autoimmune nature of a disease. A growing number of studies have begun to provide proof of the pathogenicity of NSA-Abs and insights into their pathogenic mechanisms through passive transfer or, more rarely, through active immunization animal models. Moreover, the increasing evidence that NSA-Abs in the maternal circulation can reach the fetal brain parenchyma during gestation, causing long-term effects, has led to models of antibody-induced neurodevelopmental disorders. This review summarizes different methodological approaches and the results of the animal models of N-methyl-d-aspartate receptor (NMDAR), leucine-rich glioma-inactivated 1 (LGI1), contactin-associated protein 2 (CASPR2), and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) antibody-mediated disorders and discuss the results and the limitations. We also summarize recent experiments that demonstrate that maternal antibodies to NMDAR and CASPR2 can alter development in the offspring with potential lifelong susceptibility to neurological or psychiatric disorders.


Use, tolerability and compliance of spironolactone in the treatment of heart failure.
Risk of morbidity and mortality in patients with severe heart failure (HF) is reduced by blockade of aldosterone receptors with spironolactone. However, benefits of spironolactone are potentially limited by treatment compliance and adverse events profile. The aim of this study was to estimate use of spironolactone by patients with HF, incidence of key adverse events, and patient compliance. This study was performed using data from the Quebec provincial medical and drug plans (Régie de l'Assurance Maladie du Québec, RAMQ) for patients who had a diagnosis of HF. Relative incidence of gynecomastia and hyperkalemia was estimated for users and non-users of spironolactone. Treatment adherence was estimated for users of spironolactone and compared to adherence with angiotensin converting enzyme (ACE) inhibitors, beta-blockers (β-blockers), and angiotensin receptor blockers (ARBs). RAMQ data were obtained for a total of 82,018 patients with a diagnosis of HF. Of these patients, 59.9% used an ACE inhibitor, 59.5% used a beta-blocker, 28.4% used an ARB, and 15.1% (n = 12,344) used spironolactone. Despite underestimation due to limitation of the database, the documented incidence of hyperkalemia (3.3% versus 1.4%) and gynecomastia (1.8% versus 0.7%) was significantly higher in spironolactone users than non-users (p < 0.001). Treatment compliance was significantly lower with spironolactone compared to ACE inhibitors, β-blockers, and ARBs (45.6% versus 56.1%, 59.7%, and 57.0%, respectively; p < 0.001). Persistence to treatment over a one-year period was also lower with spironolactone compared to ACE inhibitors, β-blockers, and ARBs (50.7% versus 64.5%, 70.4%, and 66.3%, respectively; p < 0.001). Use of spironolactone is associated with an incidence of adverse events, which may have an impact on treatment compliance.


Habituation of auditory responses in young autistic and neurotypical children.
Prior studies suggest that habituation of sensory responses is reduced in autism and that diminished habituation could be related to atypical autistic sensory experiences, for example, by causing brain responses to aversive stimuli to remain strong over time instead of being suppressed. While many prior studies exploring habituation in autism have repeatedly presented identical stimuli, other studies suggest group differences can still be observed in habituation to intermittent stimuli. The present study explored habituation of electrophysiological responses to auditory complex tones of varying intensities (50-80 dB SPL), presented passively in an interleaved manner, in a well-characterized sample of 127 autistic (MDQ = 65.41, SD = 20.54) and 79 typically developing (MDQ = 106.02, SD = 11.50) children between 2 and 5 years old. Habituation was quantified as changes in the amplitudes of single-trial responses to tones of each intensity over the course of the experiment. Habituation of the auditory N2 response was substantially reduced in autistic participants as compared to typically developing controls, although diagnostic groups did not clearly differ in habituation of the P1 response. Interestingly, the P1 habituated less to loud 80 dB sounds than softer sounds, whereas the N2 habituated less to soft 50 dB sounds than louder sounds. No associations were found between electrophysiological habituation and cognitive ability or participants' caregiver-reported sound tolerance (Sensory Profile Hyperacusis Index). The results present study results extend prior research suggesting habituation of certain sensory responses is reduced in autism; however, they also suggest that habituation differences observed using this study's paradigm might not be a primary driver of autistic participants' real-world sound intolerance.


Normative values for corneal nerve morphology assessed using corneal confocal microscopy: a multinational normative data set.
Corneal confocal microscopy is a novel diagnostic technique for the detection of nerve damage and repair in a range of peripheral neuropathies, in particular diabetic neuropathy. Normative reference values are required to enable clinical translation and wider use of this technique. We have therefore undertaken a multicenter collaboration to provide worldwide age-adjusted normative values of corneal nerve fiber parameters. A total of 1,965 corneal nerve images from 343 healthy volunteers were pooled from six clinical academic centers. All subjects underwent examination with the Heidelberg Retina Tomograph corneal confocal microscope. Images of the central corneal subbasal nerve plexus were acquired by each center using a standard protocol and analyzed by three trained examiners using manual tracing and semiautomated software (CCMetrics). Age trends were established using simple linear regression, and normative corneal nerve fiber density (CNFD), corneal nerve fiber branch density (CNBD), corneal nerve fiber length (CNFL), and corneal nerve fiber tortuosity (CNFT) reference values were calculated using quantile regression analysis. There was a significant linear age-dependent decrease in CNFD (-0.164 no./mm(2) per year for men, P < 0.01, and -0.161 no./mm(2) per year for women, P < 0.01). There was no change with age in CNBD (0.192 no./mm(2) per year for men, P = 0.26, and -0.050 no./mm(2) per year for women, P = 0.78). CNFL decreased in men (-0.045 mm/mm(2) per year, P = 0.07) and women (-0.060 mm/mm(2) per year, P = 0.02). CNFT increased with age in men (0.044 per year, P < 0.01) and women (0.046 per year, P < 0.01). Height, weight, and BMI did not influence the 5th percentile normative values for any corneal nerve parameter. This study provides robust worldwide normative reference values for corneal nerve parameters to be used in research and clinical practice in the study of diabetic and other peripheral neuropathies.


Dietary Animal-derived L-Arginine Intakes and Risk of Chronic Kidney Disease: a 6-year Follow-up of Tehran Lipid and Glucose Study.
There is inconsistent evidence regarding the potential role of L-arginine intake on kidney function. This study investigated the association of dietary L-arginine intake and the risk of chronic kidney disease (CKD) in adults. We evaluated 1780 men and women participated in the Tehran Lipid and Glucose Study, followed for a median of 6.3 years. Dietary intakes of total L-arginine as well as animal- and plant-derived L-arginine were assessed using the validated semi-quantitative food frequency questionnaire, at baseline. Demographics, anthropometrics, and biochemical variables were evaluated at baseline and again after a 3-year and a 6-year follow-up. The incidence of CKD was assessed across tertiles of L-arginine and its categories using multivariable logistic regression models. The mean dietary intakes of total, plant-derived, and animal-derived L-arginine were 4.1 ± 1.5 g/d, 1.8 ± 0.9 g/d, and 2.1 ± 0.8 g/d, respectively. In the fully-adjusted logistic regression model, the highest compared to the lowest intakes of animal-derived L-arginine (2.57 g/d versus 1.05 g/d) increased the risk of CKD (relative risk, 1.54; 95% confidence interval, 1.06 to 2.14, P = .02 for trend). Animal-derived L-arginine was negatively associated with changes of estimated glomerular filtration rate and creatinine clearance rate during the follow-up. There was no significant association between total or plant-derived L-arginine intakes and the risk of CKD after 6.3 years of follow-up. Our findings suggested an adverse effect of higher intakes of L-arginine from animal sources that could be a dietary risk factor for development of kidney disease.


Uterine activity during the third stage of labor.
The purpose of this study was to evaluate the use of intrauterine pressure catheters (IUPCs) to measure uterine activity during the third stage of labor. The study was conducted in a tertiary-care center. All births were attended by certified nurse-midwives. Low-risk pregnant women at term gestation had intrauterine monitoring attempted during the third stage of labor. Data were analyzed using descriptive statistics and analysis of variance. Of the 36 women in the study, 19 had clear contractions recorded, 4 had unclear contractions, and 13 had flat tracings. The IUPC was useful for measuring uterine activity in a small number of women during the third stage of labor. When measured, uterine contractions were variable in strength and frequency. Intrauterine pressure catheters can be used as a measurement of uterine activity during the third stage of labor and should be considered for future research about third-stage physiology. Further work is needed to determine the best procedure for placement, and IUPC use before expulsion of the placenta will not be useful in all women. Effective measurement of uterine activity may guide future research on interventions to reduce the risk of postpartum hemorrhage.


A Perspective on the Integration of eHealth in Treatment of Offenders: Combining Technology and the Risk-Need-Responsivity Model.
While there are multiple ways in which eHealth interventions such as online modules, apps and virtual reality can improve forensic psychiatry, uptake in practice is low. To overcome this problem, better integration of eHealth in treatment is necessary. In this perspective paper, we describe how the possibilities of eHealth can be connected to the risk-need-responsivity (RNR) model. To account for the risk-principle, stand-alone eHealth interventions might be used to offer more intensive treatment to high-risk offenders. The need-principle can be addressed by connecting novel experience-based interventions such as VR and apps to stable and acute dynamic risk factors. Finally, using and combining personalized interventions is in line with the responsivity-principle. Based on research inside and outside of forensic psychiatry, we conclude that there are many possibilities for eHealth to improve treatment-not just based on RNR, but also on other models. However, there is a pressing need for more development, implementation and evaluation research.


Respiratory Protective Equipment for Healthcare Providers During Coronavirus Pandemic: "Nec Temere, Nec Timide".
In otorhinolaryngology-head and neck surgery, there are several routine and surgical procedures applied to the upper airway that generate droplets and/or aerosols. Therefore, otorhinolaryngology-head and neck surgeons are at higher risk of being exposed to viral content. The COVID-19 pandemic has shaken the world with approximately 30 million affected cases and more than 900.000 deaths officially reported in more than 200 countries/regions from March 11th, 2020 to date (September 12th, 2020). All healthcare providers working at the frontlines of the fight against the COVID-19 are at risk of contracting the virus. In this review, we discuss the efficacy of the different types of respiratory protective equipment and remind about the surgery-based respiratory protection strategies in otorhinolaryngology and head and neck surgeries in the light of the latest literature.


Contextual effect of income inequality on birth outcomes.
Though associations between income inequality and birth outcome have been suggested, mechanisms underlying this relationship are not known. In this analysis, we examined the relationship between income inequality and preterm birth (PTB) and post-neonatal mortality (PNM) to explore two potential mechanisms-the proposed psychosocial stress and neo-material pathways. Data on singleton births from 1998 to 2000 were obtained from the CDC's National Center for Health Statistics' Linked Birth and Infant Death files. The Gini Index was utilized to measure income inequality and was divided into tertiles representing high, medium, and low county-level inequality. To determine the association between the birth outcomes and county income inequality and to account for clustering within counties, we employed generalized estimating equation (GEE) modelling. PTB increased from 8.3% in counties with low income inequality to 10.0% in counties with high inequality. The Gini Index remained modestly associated with PTB after adjusting for individual level variables and mean county-level per capita income within the total population (AOR: 1.06; 95% CI 1.03-1.09) as well as within most of the racial/ethnic groups. PNM increased from 1.15 deaths per 1000 live births in low inequality counties to 1.32 in high-inequality counties. However, after adjustment, income inequality was only associated with PNM within the non-Hispanic black population (AOR: 1.20; 95% CI 1.03-1.39). These findings may provide some support for the association between income inequality and PTB. Further research is required to elucidate the biological mechanisms of income inequality.


Sex hormones and tumor promotion in liver.
Epidemiological and experimental data strongly support a causal relationship between exposure to excessive levels of estrogens and the development of cancer in various tissues. In this paper, we have presented background information that shows a correlation between the prolonged use of oral contraceptives and the development of liver cancer. The clinical data supported the hypothesis that the estrogenic components of oral contraceptives were promoters of hepatocarcinogenesis, and the experimental evidence in support of this hypothesis and bearing on the mechanisms involved are also reviewed. The effects of estrogens on liver neoplasia and growth are: (i) synthetic steroidal estrogens are potent promoters of hepatocarcinogenesis in female rats; (ii) these estrogens stimulate liver growth at doses that are not hepatotoxic; (iii) the mechanisms by which the estrogens stimulate liver growth are indirect and include the enhancement of a serum/plasma growth factor, co-mitogenic effects which result in enhanced responsiveness of cultured hepatocytes to epidermal growth factor and decreased sensitivity of hepatocytes to growth inhibition by transforming growth factor-beta; (iv) the co-mitogenic effects of synthetic estrogens extend to endogenous estrogens and natural product estrogens; and (v) the co-mitogenic effects of estrogens for epidermal growth factor are associated with increased epidermal growth factor receptor protein levels caused by an increase in the half-life of the receptor protein. The synthetic estrogens also have weak "complete" carcinogenic activity in rat liver and strong complete carcinogenic activity in Syrian hamster kidney and Armenian hamster liver. Evidence from the literature is presented in support of a hypothesis that this process may involve indirect genotoxicity mediated through redox cycling and the formation of hydroxylated DNA bases. This process, together with the potent promoting activity of these estrogenic chemicals, may account for their complete carcinogenicity.


DispHred: A Server to Predict pH-Dependent Order-Disorder Transitions in Intrinsically Disordered Proteins.
The natively unfolded nature of intrinsically disordered proteins (IDPs) relies on several physicochemical principles, of which the balance between a low sequence hydrophobicity and a high net charge appears to be critical. Under this premise, it is well-known that disordered proteins populate a defined region of the charge-hydropathy (C-H) space and that a linear boundary condition is sufficient to distinguish between folded and disordered proteins, an approach widely applied for the prediction of protein disorder. Nevertheless, it is evident that the C-H relation of a protein is not unalterable but can be modulated by factors extrinsic to its sequence. Here, we applied a C-H-based analysis to develop a computational approach that evaluates sequence disorder as a function of pH, assuming that both protein net charge and hydrophobicity are dependent on pH solution. On that basis, we developed DispHred, the first pH-dependent predictor of protein disorder. Despite its simplicity, DispHred displays very high accuracy in identifying pH-induced order/disorder protein transitions. DispHred might be useful for diverse applications, from the analysis of conditionally disordered segments to the synthetic design of disorder tags for biotechnological applications. Importantly, since many disorder predictors use hydrophobicity as an input, the here developed framework can be implemented in other state-of-the-art algorithms.


Reconstruction of the Heel, Middle Foot Sole, and Plantar Forefoot with the Medial Plantar Artery Perforator Flap: Clinical Experience with 28 Cases.
Defects in the weight-bearing region of the foot sole can represent a substantial restriction in quality of life and pose a challenge for reconstructive plastic surgery. The purpose of this article is to report the authors' experience with the use of the medial plantar artery perforator flap for reconstruction of defects of the foot sole in three different regions: heel, middle foot sole, and plantar forefoot. From January of 2003 to May of 2016, 28 patients (13 male and 15 female patients) with an average age of 54 years (range, 12 to 84 years) underwent reconstruction with 28 medial plantar artery perforator flaps. Twenty-six flaps were harvested as pedicle perforator flaps and two as free perforator flaps. All flaps were raised from the ipsilateral instep area. The defect locations included the heel (20 cases), middle foot sole (four cases), and forefoot (four cases). The causes of reconstruction were tumors in 18 patients, decubitus in eight patients, and trauma in two patients. The flap sizes varied from 2.5 × 2.5 cm to 5.5 × 9.5 cm. All of the flaps survived completely after surgery, apart from one. The donor sites were all covered with a split-thickness skin graft. Follow-up observations were conducted for 4 to 12 months, and all patients had good functional recovery with satisfactory cosmetic results. The medial plantar artery perforator flap can be considered an optimal method of foot sole reconstruction not only for covering the weight-bearing area of the heel but also for the middle and forefoot plantar region. Therapeutic, IV.


TLR7- and TLR9-responsive human B cells share phenotypic and genetic characteristics.
B cells activated by nucleic acid-sensing TLR7 and TLR9 proliferate and secrete immune globulins. Memory B cells are presumably more responsive due to higher TLR expression levels, but selectivity and differential outcomes remain largely unknown. In this study, peripheral blood human B cells were stimulated by TLR7 or TLR9 ligands, with or without IFN-α, and compared with activators CD40L plus IL-21, to identify differentially responsive cell populations, defined phenotypically and by BCR characteristics. Whereas all activators induced differentiation and Ab secretion, TLR stimulation expanded IgM(+) memory and plasma cell lineage committed populations, and favored secretion of IgM, unlike CD40L/IL-21, which drove IgM and IgG more evenly. Patterns of proliferation similarly differed, with CD40L/IL-21 inducing proliferation of most memory and naive B cells, in contrast with TLRs that induced robust proliferation in a subset of these cells. On deep sequencing of the IgH locus, TLR-responsive B cells shared patterns of IgHV and IgHJ usage, clustering apart from CD40L/IL-21 and control conditions. TLR activators, but not CD40L/IL-21, similarly promoted increased sharing of CDR3 sequences. TLR-responsive B cells were characterized by more somatic hypermutation, shorter CDR3 segments, and less negative charges. TLR activation also induced long positively charged CDR3 segments, suggestive of autoreactive Abs. Testing this, we found culture supernatants from TLR-stimulated B cells to bind HEp-2 cells, whereas those from CD40L/IL-21-stimulated cells did not. Human B cells possess selective sensitivity to TLR stimulation, with distinctive phenotypic and genetic signatures.


Superionic and metallic states of water and ammonia at giant planet conditions.
The phase diagrams of water and ammonia were determined by constant pressure ab initio molecular dynamic simulations at pressures (30 to 300 gigapascal) and temperatures (300 to 7000 kelvin) of relevance for the middle ice layers of the giant planets Neptune and Uranus. Along the planetary isentrope water and ammonia behave as fully dissociated ionic, electronically insulating fluid phases, which turn metallic at temperatures exceeding 7000 kelvin for water and 5500 kelvin for ammonia. At lower temperatures, the phase diagrams of water and ammonia exhibit a superionic solid phase between the solid and the ionic liquid. These simulations improve our understanding of the properties of the middle ice layers of Neptune and Uranus.


[Cumulative incidence, comorbility and disability for anxiety disorders in patients of a mutual worker accident.]
Neurotic disorders are one of the main health problems of Western societies. Its impact on labor productivity grows exponentially with an increase in its incidence and the duration of temporary disability (TD) processes. To know the characteristics of the patients belonging to a mutual of work accidents who had transient disability due to neurotic disorders, their temporal and geographic distribution, their comorbidities and those variables that affected their duration. Retrospective descriptive study of the workers protected from a mutual of work accidents, national scope, who studied a disability period between 2006-2016 with diagnoses included between codes 300 and 300.9 of the CIE-9-CM, corresponding to Neurotic Disorders. We analyze clinical-demographic variables of patients, comorbidities during studied period and distributions by year and province, by Bayesian inference. We performed a logistic regression for the dependent variable duration of the TD adjusted for age, sex, toxic consumption, consultations, year, regulatory base and payment type. We registered 56,619 processes in Spain, which corresponded to 1.5% of the total TD processes served in the period. The highest percentage of cases was grouped between 2007 and 2009, when 11% of the population were registered annually. The variables associated with an increase in this duration were: older age, male sex, psychiatric and psychological consultations, diagnostic year, regulatory base (whose average was 50.5 euros) and type of payment (delegate in 78.2% of cases and direct in 21.8%). The prevalence by provinces was more marked in the north and the islands (Barcelona, Lleida, Las Palmas, Islas Baleares, Coruña, Cantabria, Girona, Álava, Tarragona, Pontevedra y Asturias they have more than 6%).The durations of the disability were greater in the east of the country. The most prevalent comorbidities during the study period were low back pain (with an average of 65.1 days of IT anxiety), cervical pain (with an average of 67 days) and digestive disorders (with an average of 59.4 days). There are comorbidities and variables associated with the temporary disability derived from neurotic disorders, with geographical differences. Deepening their knowledge and impact could promote better preventive and therapeutic approaches that allow an earlier functional recovery.


Building Resilient Cities: Climate Change and Health Interlinkages in the Planning of Public Spaces.
Greenhouse gases emissions resulting from the combustion of fossil fuels are worsening air quality and affecting the climate system. While climate change impacts on meteorological variables affects air quality by altering the concentration and distribution of pollutants, air pollution significantly influences the climate, leading to negative impacts on human health. Due to the combination of high temperatures, air pollution, and high population density, cities are particularly vulnerable to climate change impacts. The planning and design of public spaces aimed at climate change mitigation and adaptation can result in multiple co-benefits for human health, while reducing social inequalities. To address the major research gaps in the communication between health and planning experts, and the lack of capacity among public sectors and policy makers, it is necessary to promote capacity building and knowledge sharing between the planning and health sectors. The purpose of this article is to develop preliminary recommendations for a process that allows a comprehensive assessment of the interlinkages between climate and health, social, environmental, and economic vulnerabilities, and the quality of the urban spaces, to support local governments, policymakers, and education institutions in making informed decisions for public spaces. The methods applied were a literature review and interviews with experts.


Microparticle-Induced Coagulation Relates to Coronary Artery Atherosclerosis in Severe Aortic Valve Stenosis.
Circulating microparticles (MPs) derived from endothelial cells and blood cells bear procoagulant activity and promote thrombin generation. Thrombin exerts proinflammatory effects mediating the progression of atherosclerosis. Aortic valve stenosis may represent an atherosclerosis-like process involving both the aortic valve and the vascular system. The aim of this study was to investigate whether MP-induced thrombin generation is related to coronary atherosclerosis and aortic valve calcification. In a cross-sectional study of 55 patients with severe aortic valve stenosis, we assessed the coronary calcification score (CAC) as indicator of total coronary atherosclerosis burden, and aortic valve calcification (AVC) by computed tomography. Thrombin-antithrombin complex (TATc) levels were measured as a marker for thrombin formation. Circulating MPs were characterized by flow cytometry according to the expression of established surface antigens and by measuring MP-induced thrombin generation. Patients with CAC score below the median were classified as patients with low CAC, patients with CAC Score above the median as high CAC. In patients with high CAC compared to patients with low CAC we detected higher levels of TATc, platelet-derived MPs (PMPs), endothelial-derived MPs (EMPs) and MP-induced thrombin generation. Increased level of PMPs and MP-induced thrombin generation were independent predictors for the severity of CAC. In contrast, AVC Score did not differ between patients with high and low CAC and did neither correlate with MPs levels nor with MP-induced thrombin generation. In patients with severe aortic valve stenosis MP-induced thrombin generation was independently associated with the severity of CAC but not AVC indicating different pathomechanisms involved in coronary artery and aortic valve calcification.


Effect of site selection on pain of intravenous cannula insertion: A prospective randomised study.
Pain on intravenous (IV) cannulation continues to cause considerable anxiety among the patients visiting the hospital for elective surgery. Often, it is the only unpleasant experience, especially in ambulatory surgical settings. Although, anecdotal evidence suggests that antecubital fossa (ACF) might be less painful site for venous cannulation, no scientific study exists to validate the same. In this prospective randomised study, effect of site selection on pain of venous cannulation was studied. Fifty-five consecutive adults, scheduled to undergo elective surgery, were randomly allocated to get IV cannulation first on ACF (28 patients) or on dorsum of hand (DOH) (27 patients) followed by cannulation on the contralateral arm on the alternative site (DOH or ACF). Five patients were excluded due to multiple cannulation attempts. Pain scores on cannulation related to both sites were recorded and compared. Non-parametric data and frequency data analysis, using the Wilcoxon signed rank test or the Chi-square test as appropriate, showed that ACF approach was significantly less painful in comparison to the DOH when using a 20-gauge cannula for venous cannulation (P < 0.05). We recommend that in the absence of any contraindications, ACF should be the cannulation site of choice. However, considerations like increased chance of kinking and obstruction might preclude such practice.