Anomalous intrathoracic left vagus and recurrent laryngeal nerve course.
An anomalous course of the vagus nerve is an extremely rare congenital anomaly. We report the case of a 59-year-old man who had left lung cancer. We performed a left upper lobectomy and mediastinal lymphadenectomy, paying attention to the pathway of the left vagus nerve, which had an anomalous course passing anteriorly to the left pulmonary trunk between the apical and anterior branches. The left recurrent laryngeal nerve hooked around the lower border of the apical branch of the left pulmonary artery.


Epigenetic Histone Methylation of PPARγ and CPT1A Signaling Contributes to Betahistine Preventing Olanzapine-Induced Dyslipidemia.
As a partial histamine H1 receptor agonist and H3 antagonist, betahistine has been reported to partially prevent olanzapine-induced dyslipidemia and obesity through a combination therapy, although the underlying epigenetic mechanisms are still not known. Recent studies have revealed that histone regulation of key genes for lipogenesis and adipogenesis in the liver is one of the crucial mechanisms for olanzapine-induced metabolic disorders. This study investigated the role of epigenetic histone regulation in betahistine co-treatment preventing dyslipidemia and fatty liver caused by chronic olanzapine treatment in a rat model. In addition to abnormal lipid metabolism, the upregulation of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein (C/EBPα), as well as the downregulation of carnitine palmitoyltransferase 1A (CPT1A) in the liver induced by olanzapine, were significantly attenuated by betahistine co-treatment. In addition, betahistine co-treatment significantly enhanced the global expression of H3K4me and the enrichment of H3K4me binding on the promoter of Cpt1a gene as revealed by ChIP-qPCR, but inhibited the expression of one of its site-specific demethylases, lysine (K)-specific demethylase 1A (KDM1A). Betahistine co-treatment also significantly enhanced the global expression of H3K9me and the enrichment of H3K9me binding on the promoter of the Pparg gene, but inhibited the expression of two of its site-specific demethylases, lysine demethylase 4B (KDM4B) and PHD finger protein 2 (PHF2). These results suggest that betahistine attenuates abnormal adipogenesis and lipogenesis triggered by olanzapine through modulating hepatic histone methylation, and thus inhibiting the PPARγ pathway-mediated lipid storage, while at the same time promoting CP1A-mediated fatty acid oxidation.


Respiratory Motion Mitigation and Repeatability of Two Diffusion-Weighted MRI Methods Applied to a Murine Model of Spontaneous Pancreatic Cancer.
Respiratory motion and increased susceptibility effects at high magnetic fields pose challenges for quantitative diffusion-weighted MRI (DWI) of a mouse abdomen on preclinical MRI systems. We demonstrate the first application of radial k-space-sampled (RAD) DWI of a mouse abdomen using a genetically engineered mouse model of pancreatic ductal adenocarcinoma (PDAC) on a 4.7 T preclinical scanner equipped with moderate gradient capability. RAD DWI was compared with the echo-planar imaging (EPI)-based DWI method with similar voxel volumes and acquisition times over a wide range of b-values (0.64, 535, 1071, 1478, and 2141 mm2/s). The repeatability metrics are assessed in a rigorous test-retest study (n = 10 for each DWI protocol). The four-shot EPI DWI protocol leads to higher signal-to-noise ratio (SNR) in diffusion-weighted images with persisting ghosting artifacts, whereas the RAD DWI protocol produces relatively artifact-free images over all b-values examined. Despite different degrees of motion mitigation, both RAD DWI and EPI DWI allow parametric maps of apparent diffusion coefficients (ADC) to be produced, and the ADC of the PDAC tumor estimated by the two methods are 1.3 ± 0.24 and 1.5 ± 0.28 × 10-3 mm2/s, respectively (p = 0.075, n = 10), and those of a water phantom are 3.2 ± 0.29 and 2.8 ± 0.15 × 10-3 mm2/s, respectively (p = 0.001, n = 10). Bland-Altman plots and probability density function reveal good repeatability for both protocols, whose repeatability metrics do not differ significantly. In conclusion, RAD DWI enables a more effective respiratory motion mitigation but lower SNR, while the performance of EPI DWI is expected to improve with more advanced gradient hardware.


Nonparametric Coupled Bayesian Dictionary and Classifier Learning for Hyperspectral Classification.
We present a principled approach to learn a discriminative dictionary along a linear classifier for hyperspectral classification. Our approach places Gaussian Process priors over the dictionary to account for the relative smoothness of the natural spectra, whereas the classifier parameters are sampled from multivariate Gaussians. We employ two Beta-Bernoulli processes to jointly infer the dictionary and the classifier. These processes are coupled under the same sets of Bernoulli distributions. In our approach, these distributions signify the frequency of the dictionary atom usage in representing class-specific training spectra, which also makes the dictionary discriminative. Due to the coupling between the dictionary and the classifier, the popularity of the atoms for representing different classes gets encoded into the classifier. This helps in predicting the class labels of test spectra that are first represented over the dictionary by solving a simultaneous sparse optimization problem. The labels of the spectra are predicted by feeding the resulting representations to the classifier. Our approach exploits the nonparametric Bayesian framework to automatically infer the dictionary size-the key parameter in discriminative dictionary learning. Moreover, it also has the desirable property of adaptively learning the association between the dictionary atoms and the class labels by itself. We use Gibbs sampling to infer the posterior probability distributions over the dictionary and the classifier under the proposed model, for which, we derive analytical expressions. To establish the effectiveness of our approach, we test it on benchmark hyperspectral images. The classification performance is compared with the state-of-the-art dictionary learning-based classification methods.


A recurrent fuzzy-neural model for dynamic system identification.
This paper presents a fuzzy modeling approach for identification of dynamic systems. In particular, a new fuzzy model, the Dynamic Fuzzy Neural Network (DFNN), consisting of recurrent TSK rules, is developed. The premise and defuzzification parts are static while the consequent parts of the fuzzy rules are recurrent neural networks with internal feedback and time delay synapses. The network is trained by means of a novel learning algorithm, named Dynamic-Fuzzy Neural Constrained Optimization Method (D-FUNCOM), based on the concept of constrained optimization. The proposed algorithm is general since it can be applied to locally as well as fully recurrent networks, regardless of their structures. An adaptation mechanism of the maximum parameter change is presented as well. The proposed dynamic model, equipped with the learning algorithm, is applied to several temporal problems, including modeling of a NARMA process and the noise cancellation problem. Performance comparisons are conducted with a series of static and dynamic systems and some existing recurrent fuzzy models. Simulation results show that DFNN compares favorably with its competing rivals and thus it can be considered for efficient system identification.


Elucidating the Key Determinants of Structure, Folding, and Stability for the ( 4β+ α ) Conformation of the B1 Domain of Protein G Using Bioinformatics Approaches.
The B1 domain of protein G (GB1) is a small, 56 amino acid bacterial immunoglobulin-binding protein with a 4β+ α fold. Architecturally, it is composed of a two-layer sandwich consisting of a four-stranded β -sheet that packs against an α -helix. Using several bioinformatics approaches, we investigated which residues may be key determinants of this fold. We identified nine structurally conserved amino acids using a conservation analysis and propose they are critical to forming and stabilizing the fold. The nine conserved residues form a predominantly hydrophobic nucleus within the core of GB1. A network analysis of all the long-range interactions in the structure of GB1 in concert with a betweenness centrality analysis revealed the relative significance of each conserved amino acid residue based on the number and location of the interactions. This bioinformatics analysis provides an important foundation for the design and interpretation of both computational and experimental work which may be helpful in solving the protein folding problem.


Efficient recombinant production in mammalian cells using a novel IR/MAR gene amplification method.
We previously found that plasmids bearing a mammalian replication initiation region (IR) and a nuclear matrix attachment region (MAR) efficiently initiate gene amplification and spontaneously increase their copy numbers in animal cells. In this study, this novel method was applied to the establishment of cells with high recombinant antibody production. The level of recombinant antibody expression was tightly correlated with the efficiency of plasmid amplification and the cytogenetic appearance of the amplified genes, and was strongly dependent on cell type. By using a widely used cell line for industrial protein production, CHO DG44, clones expressing very high levels of antibody were easily obtained. High-producer clones stably expressed the antibody over several months without eliciting changes in both the protein expression level and the cytogenetic appearance of the amplified genes. The integrity and reactivity of the protein produced by this method was fine. In serum-free suspension culture, the specific protein production rate in high-density cultures was 29.4 pg/cell/day. In conclusion, the IR/MAR gene amplification method is a novel and efficient platform for recombinant antibody production in mammalian cells, which rapidly and easily enables the establishment of stable high-producer cell clone.


Regulated Cell Death as a Therapeutic Target for Novel Antifungal Peptides and Biologics.
The rise of microbial pathogens refractory to conventional antibiotics represents one of the most urgent and global public health concerns for the 21st century. Emergence of Candida auris isolates and the persistence of invasive mold infections that resist existing treatment and cause severe illness has underscored the threat of drug-resistant fungal infections. To meet these growing challenges, mechanistically novel agents and strategies are needed that surpass the conventional fungistatic or fungicidal drug actions. Host defense peptides have long been misunderstood as indiscriminant membrane detergents. However, evidence gathered over the past decade clearly points to their sophisticated and selective mechanisms of action, including exploiting regulated cell death pathways of their target pathogens. Such peptides perturb transmembrane potential and mitochondrial energetics, inducing phosphatidylserine accessibility and metacaspase activation in fungi. These mechanisms are often multimodal, affording target pathogens fewer resistance options as compared to traditional small molecule drugs. Here, recent advances in the field are examined regarding regulated cell death subroutines as potential therapeutic targets for innovative anti-infective peptides against pathogenic fungi. Furthering knowledge of protective host defense peptide interactions with target pathogens is key to advancing and applying novel prophylactic and therapeutic countermeasures to fungal resistance and pathogenesis.


A negative trial of inpatient geriatric consultation. Lessons learned and recommendations for future research.
To determine the effectiveness of inpatient interdisciplinary geriatric consultation provided during hospitalization to frail, elderly subjects. SUBJECTS AND SITE: Admission cohort of 197 men admitted from 1985 through 1989, aged 65 years or more, meeting proxy criteria for frailty, living within follow-up area, without terminal illness, and without prolonged nursing home residence. Randomized controlled trial of inpatient geriatric consultation at a tertiary care Veterans Affairs hospital. Differences were determined between groups in the Physical Self-Maintenance Scale, Instrumental Activities of Daily Living, Mini-Mental State Examination, Morale Scale, and nursing home and health care utilization. No differences were seen between groups in any measure after the intervention or during 1 year of follow-up. Intervention implementation may have been incomplete due to compliance and resource availability. This trial is not definitive in determining whether geriatric consultation is effective or ineffective. Lessons learned from this research indicate that future studies should target frail subjects, include intervention-specific measures, and be conducted with direct control of comprehensive resources.


DYRK1A and glycogen synthase kinase 3beta, a dual-kinase mechanism directing proteasomal degradation of CRY2 for circadian timekeeping.
Circadian molecular oscillation is generated by a transcription/translation-based feedback loop in which CRY proteins play critical roles as potent inhibitors for E-box-dependent clock gene expression. Although CRY2 undergoes rhythmic phosphorylation in its C-terminal tail, structurally distinct from the CRY1 tail, little is understood about how protein kinase(s) controls the CRY2-specific phosphorylation and contributes to the molecular clockwork. Here we found that Ser557 in the C-terminal tail of CRY2 is phosphorylated by DYRK1A as a priming kinase for subsequent GSK-3beta (glycogen synthase kinase 3beta)-mediated phosphorylation of Ser553, which leads to proteasomal degradation of CRY2. In the mouse liver, DYRK1A kinase activity toward Ser557 of CRY2 showed circadian variation, with its peak in the accumulating phase of CRY2 protein. Knockdown of Dyrk1a caused abnormal accumulation of cytosolic CRY2, advancing the timing of a nuclear increase of CRY2, and shortened the period length of the cellular circadian rhythm. Expression of an S557A/S553A mutant of CRY2 phenocopied the effect of Dyrk1a knockdown in terms of the circadian period length of the cellular clock. DYRK1A is a novel clock component cooperating with GSK-3beta and governs the Ser557 phosphorylation-triggered degradation of CRY2.


A multi-centre phase IIa clinical study of predictive testing for preeclampsia: improved pregnancy outcomes via early detection (IMPROvED).
5% of first time pregnancies are complicated by pre-eclampsia, the leading cause of maternal death in Europe. No clinically useful screening test exists; consequentially clinicians are unable to offer targeted surveillance or preventative strategies. IMPROvED Consortium members have pioneered a personalised medicine approach to identifying blood-borne biomarkers through recent technological advancements, involving mapping of the blood metabolome and proteome. The key objective is to develop a sensitive, specific, high-throughput and economically viable early pregnancy screening test for pre-eclampsia. We report the design of a multicentre, phase IIa clinical study aiming to recruit 5000 low risk primiparous women to assess and refine innovative prototype tests based on emerging metabolomic and proteomic technologies. Participation involves maternal phlebotomy at 15 and 20 weeks' gestation, with optional testing and biobanking at 11 and 34 weeks. Blood samples will be analysed using two innovative, proprietary prototype platforms; one metabolomic based and one proteomic based, both of which outperform current biomarker based screening tests at comparable gestations. Analytical and clinical data will be collated and analysed via the Copenhagen Trials Unit. The IMPROvED study is expected to refine proteomic and metabolomic panels, combined with clinical parameters, and evaluate clinical applicability as an early pregnancy predictive test for pre-eclampsia. If 'at risk' patients can be identified, this will allow stratified care with personalised fetal and maternal surveillance, early diagnosis, timely intervention, and significant health economic savings. The IMPROvED biobank will be accessible to the European scientific community for high quality research into the cause and prevention of adverse pregnancy outcome. Trial registration number NCT01891240The IMPROvED project is funded by the seventh framework programme for Research and Technological development of the EU. http://www.fp7-improved.eu/


A rat model for sickle cell-mediated vaso-occlusion in retina.
Occlusions of the retinal vasculature are the initiating event in sickle cell retinopathy. In order to understand the mechanism(s) of sickle cell-mediated occlusion, a rat model was developed. Red blood cells (RBCs) from patients homozygous for hemoglobin (Hb) S (SS) or double heterozygous for Hb S and Hb C (SC) were separated on Percoll-Larex continuous density gradients, labeled with fluorescein isothiocyanate (FITC), and delivered via the left ventricle to anesthetized, ventilated rats. Blood gas levels were altered by changing inspired gas and monitored via a femoral arterial catheter. After the RBCs circulated for 5 min, animals were perfused with heparinized saline, the eyes enucleated, and the retinas removed and processed by our ADPase flatmount technique. The retinal vasculature was visualized under dark-field illumination and the FITC-RBCs visualized by fluorescence microscopy. Greater numbers of high-density SS cells (SS4, which consist of dense, dehydrated discocytes and irreversible sickled cells) were retained in the normal rat retinal vasculature than normal-density SS cells (SS2, which have the same density as normal AA cells, but consist of reticulocytes and young cells). Retention of SS4 cells was inversely dependent on the arterial oxygen tension. Most SS4s were retained in capillaries, but a few were observed within precapillary arterioles. The retained RBCs occupied the full lumenal diameter of vessels in most cases. In contrast, very few RBCs from SC donors (normal or high density) were retained in the normal retinal vasculature and retention did not increase significantly with hypoxia. This model demonstrates that high-density SS cells, which include irreversibly sickled cells, are retained in normal rat retinal vessels and that the number retained is oxygen dependent. Furthermore, it appears that trapping, not adhesion, is responsible for retention of RBCs in the normal retinal vasculature because there was preferential retention of SS4 cells, which are known to have lower adherence propensity, and the retained RBCs blocked the full diameter of the vessel. These results also demonstrate that the mechanism of vascular obstruction by SS and SC RBCs is different because low retention of SC cells was observed. The well-known propensity of SC patients to have retinal abnormalities must involve extraerythrocytic factors like increased hematocrit, induction of adhesive molecules and integrins, etc.


Transmission of feline viral rhinotracheitis.
The transmission of feline viral rhinotracheitis (FVR) virus was investigated. Virus could be successfully transmitted between shedding carrier cats and unrelated susceptible kittens, but only if there was intimate contact between them. Studies on the transmission of FVR virus from carrier queens to their kittens showed that although four of 10 queens shed virus in the post partum period, a total of only four kittens from three litters developed a contact infection. All four kittens remained asymptomatic. Two shed for one day only and did not become carriers (as evidenced by corticosteroid treatment) and two shed for 15 days and 25 days and were subsequently shown to have become carriers. None of the remaining kittens tested shed virus. There was no evidence of in utero transmission between FVR-recovered queens and kittens. Passive antibody titres in kittens born to FVR-recovered queens declined to less than 1 in 4 in individual animals by two to 10 weeks of age. Mean titres calculated from a linear regression equation reached less than 1 in 4 and less than 1 in 2 by six and nine weeks of age, respectively. Experiments using a multistage liquid impinger demonstrated that FVR virus was unlikely to be transmitted between cats by aerosol and this was confirmed by the ability of a sentinel cat to withstand virus shedding from carriers over a six month period, although housed in the same air space.


Clinicians' satisfaction with a hospital blood transfusion service: a marketing analysis of a monopoly supplier.
One of the objectives of the NHS reforms is to improve customer focus within the health service. In a study to assess the quality of customer service provided by the Edinburgh and South East Scotland Blood Transfusion Service a 19 item questionnaire survey of the main clinical users of the service was performed to ascertain their satisfaction, measured on a 5 point anchored scale, with important aspects of the service, including medical consultation, diagnostic services, blood and blood components or products and their delivery, and general satisfaction with the service. Of 122 clinicians in medical and surgical disciplines in five hospitals in Edinburgh, 72 (59%) replied. Fourteen (22%) indicated dissatisfaction with any aspect of the medical consultation service, owing to inadequate follow up of clinical contacts and unsatisfactory routing of incoming calls. Diagnostic services were criticised for the presentation, communication, and interpretation of results. The restricted availability of whole blood, the necessity to order platelets and plasma through the duty blood transfusion service doctor, and the use of a group and screen policy, attracted criticism from a small number of clinicians. Ten of 68 respondents expressed dissatisfaction with delivery of blood and components to the wards and theatres. The findings indicate that the clinicians served by this blood transfusion service are largely satisfied with the service. Changes are being implemented to improve reporting of laboratory results and measures taken to improve liaison with clinicians.


Airway management in obese patients.
The well-known difficulties in airway management in obese patients are caused by obesity-related airways and respiratory changes. Anesthesiologists confront a number of troubles, including rapid oxygen desaturation, difficulty with laryngoscopy/intubation and mask ventilation, and increased susceptibility to the respiratory depressant effects of anesthetic drugs. Preoperative assessment of the airways in the obese should include examination of specific predictors of difficult mask ventilation other than those for difficult intubation. Difficulties in airway management are decreased after providing optimal preoxygenation and positioning ("ramped"). Other strategies may include availability of alternative airway management devices, including new video laryngoscopes that significantly improve the visualization of the larynx and thereby facilitate intubation. If awake intubation is mandatory, it may be performed with fibrobronchoscope after providing an adequate topical anesthesia and sedation with short-acting drugs, such as remifentanil. Succinylcholine for rapid sequence induction might be replaced by rocuronium where sugammadex is available for reversal. A complete reversal of neuromuscular block, measured by train-of-four monitoring, should be obtained before extubation, which requires a fully awake patient in the same position with airway equipment used for intubation.


Ketogenic diet sensitizes glucose control of hippocampal excitability.
A high-fat low-carbohydrate ketogenic diet (KD) is an effective treatment for refractory epilepsy, yet myriad metabolic effects in vivo have not been reconciled clearly with neuronal effects. A KD limits blood glucose and produces ketone bodies from β-oxidation of lipids. Studies have explored changes in ketone bodies and/or glucose in the effects of the KD, and glucose is increasingly implicated in neurological conditions. To examine the interaction between altered glucose and the neural effects of a KD, we fed rats and mice a KD and restricted glucose in vitro while examining the seizure-prone CA3 region of acute hippocampal slices. Slices from KD-fed animals were sensitive to small physiological changes in glucose, and showed reduced excitability and seizure propensity. Similar to clinical observations, reduced excitability depended on maintaining reduced glucose. Enhanced glucose sensitivity and reduced excitability were absent in slices obtained from KD-fed mice lacking adenosine A1 receptors (A1Rs); in slices from normal animals effects of the KD could be reversed with blockers of pannexin-1 channels, A1Rs, or KATP channels. Overall, these studies reveal that a KD sensitizes glucose-based regulation of excitability via purinergic mechanisms in the hippocampus and thus link key metabolic and direct neural effects of the KD.


Ultrasonographic evaluation of lung parenchyma involvement in sarcoidosis.
To use ultrasonography (USG) for the evaluation of lung parenchyma in patients with sarcoidosis, andto compare the USG findings with the results of a high-resolution computerized tomography (HRCT) and pulmonary function test-carbon monoxide diffusion test (PFT-DLCO), which are commonly used methods in the evaluation of parenchymal involvement in sarcoidosis. Patients with sarcoidosis and healthy controls were enrolled in the study between January 2015 and December 2017. The clinical findings, HRCT and PFT-DLCO results of all subjects were recorded, and USG findings and comet tail artifact (CTA) measurements were recorded by another pulmonologist. The USG, HRCT and SFT-DLCO findings were compared between the two groups. Based on the findings of theclinical-radiologic investigations and PFT-DLCO, as the current gold standard in diagnosis, the sensitivity and specificity of USG in demonstrating lung parenchyma involvement in sarcoidosis patients were estimated. The sarcoidosis group consisted of 79 patients and the control group included 34 subjects. The mean number of CTAs in the sarcoidosis and control groups was 33.4 and 25, respectively (p=0.001). In the sarcoidosis group, the number of CTAs in patients with DLCO% <80 and ≥80% was 37.4 and 29.7, respectively (p=0.011), and a negative correlation was identified between the number of CTAs and DLCO% (p=0.019 r=-0.267). The mean number of CTAs in patients with and without parenchymal involvement in HRCT was 36 and 25.5, respectively (p=0.001). The number of CTAs in the patients with sarcoidosis with a normal DLCO% value (≥80%) was higher than in the control group (p=0.014). The diagnostic sensitivity and specificity of thoracic USG were found to be 76% and 53%, respectively. The number of CTAs in patients with sarcoidosis was higher than that of the healthy controls. The number of CTAs in patients with sarcoidosis with parenchymal involvement in HRCT and/or a low DLCO (<80%) was also elevated. Thoracic USG has a high sensitivity (76%) in demonstrating parenchymal involvement in patients with sarcoidosis.


Protease-deficient Bacillus subtilis host strains for production of Staphylococcal protein A.
We constructed strains of Bacillus subtilis which produced very low levels of extracellular proteases. These strains carried insertion or deletion mutations in the subtilisin structural gene (apr) which were constructed in vitro by using the cloned gene. The methods used to construct the mutations involved the use of a plasmid vector which allowed the selection of chromosomal integrates and their subsequent excision by homologous recombination to effect replacement of the chromosomal apr gene by a derivative carrying an inactivating insert with a selectable marker (a cat gene conferring chloramphenicol resistance). The strains produced no subtilisin, no detectable extracellular metalloprotease activity, and residual extracellular serine protease levels as low as 0.5% of that of the standard strain from which they were derived. The strains proved to be superior host strains for the production of staphylococcal protein A, accumulating higher levels of intact protein than do previously available B. subtilis strains.


Deficiency of thioredoxin-interacting protein (TXNIP) results in age-related thrombocytopenia due to megakaryocyte oxidative stress.
Platelets are generated from megakaryocytes (MKs), mainly located in the bone marrow (BM). Megakaryopoiesis can be affected by genetic disorders, metabolic diseases, and aging. The molecular mechanisms underlying platelet count regulation have not been fully elucidated. In the present study, we investigated the role of thioredoxin-interacting protein (TXNIP), a protein that regulates cellular metabolism in megakaryopoiesis, using a Txnip-/- mouse model. Wild-type (WT) and Txnip-/- mice (2-27-month-old) were studied. BM-derived MKs were analyzed to investigate the role of TXNIP in megakaryopoiesis with age. The global transcriptome of BM-derived CD41+ megakaryocyte precursors (MkPs) of WT and Txnip-/- mice were compared. The CD34+ hematopoietic stem cells isolated from human cord blood were differentiated into MKs. Txnip-/- mice develop thrombocytopenia at 4-5 months that worsens with age. During ex vivo megakaryopoiesis, Txnip-/- MkPs remained small, with decreased levels of MK-specific markers. Critically, Txnip-/- MkPs exhibited reduced mitochondrial reactive oxygen species, of which was related to AKT activity. Txnip-/- MkPs also showed elevated glycolysis, alongside increased glucose uptake for ATP production. Total RNA-sequencing revealed enrichment for oxidative stress- and apoptosis-related genes in differentially expressed genes between Txnip-/- and WT MkPs. The effects of TXNIP on MKs were recapitulated during the differentiation of human cord blood-derived CD34+ hematopoietic stem cells. We provide evidence that the megakaryopoiesis pathway becomes exhausted with age in Txnip-/- mice with a decrease in terminal, mature MKs that response to thrombocytopenic challenge. Overall, this study demonstrates the role of TXNIP in megakaryopoiesis, regulating mitochondrial metabolism.


Disappearance of beta2-adrenergic receptors on astrocytes in canine distemper encephalitis: possible implications for the pathogenesis of multiple sclerosis.
It has been reported that astrocytes in the white matter of patients with multiple sclerosis (MS) lack beta2-adrenergic receptors. This abnormality might explain why astrocytes in active MS plaques aberrantly express major histocompatibility (MHC) class II molecules, which play an important role in the immunological cascade leading to myelin destruction. Canine distemper (CD) virus primarily infects astrocytes and causes a demyelinating disease in dogs that closely resembles MS. In control dogs, including three dogs with another inflammatory disease, beta2-adrenergic receptor immunoreactivity was observed on both neurons and astrocytes. In dogs with CD encephalitis, beta2-adrenergic receptors were present on neurons, but were absent on astrocytes in acute lesions, demyelinated lesions, and normal-appearing white matter. Similar to MS, several astrocytes in demyelinated lesions expressed MHC class II. These findings suggest that MS and the demyelinating stages of CD encephalitis have a common pathogenetic factor, and that the loss of astrocytic beta2-adrenergic receptors in MS might be induced by a viral infection of astrocytes.


Performance Evaluation of Matrix Factorization for fMRI Data.
A hypothesis in the study of the brain is that sparse coding is realized in information representation of external stimuli, which has been experimentally confirmed for visual stimulus recently. However, unlike the specific functional region in the brain, sparse coding in information processing in the whole brain has not been clarified sufficiently. In this study, we investigate the validity of sparse coding in the whole human brain by applying various matrix factorization methods to functional magnetic resonance imaging data of neural activities in the brain. The result suggests the sparse coding hypothesis in information representation in the whole human brain, because extracted features from the sparse matrix factorization (MF) method, sparse principal component analysis (SparsePCA), or method of optimal directions (MOD) under a high sparsity setting or an approximate sparse MF method, fast independent component analysis (FastICA), can classify external visual stimuli more accurately than the nonsparse MF method or sparse MF method under a low sparsity setting.


Global analysis of ribosome-associated noncoding RNAs unveils new modes of translational regulation.
Eukaryotic transcriptomes contain a major non-protein-coding component that includes precursors of small RNAs as well as long noncoding RNA (lncRNAs). Here, we utilized the mapping of ribosome footprints on RNAs to explore translational regulation of coding and noncoding RNAs in roots of Arabidopsis thaliana shifted from replete to deficient phosphorous (Pi) nutrition. Homodirectional changes in steady-state mRNA abundance and translation were observed for all but 265 annotated protein-coding genes. Of the translationally regulated mRNAs, 30% had one or more upstream ORF (uORF) that influenced the number of ribosomes on the principal protein-coding region. Nearly one-half of the 2,382 lncRNAs detected had ribosome footprints, including 56 with significantly altered translation under Pi-limited nutrition. The prediction of translated small ORFs (sORFs) by quantitation of translation termination and peptidic analysis identified lncRNAs that produce peptides, including several deeply evolutionarily conserved and significantly Pi-regulated lncRNAs. Furthermore, we discovered that natural antisense transcripts (NATs) frequently have actively translated sORFs, including five with low-Pi up-regulation that correlated with enhanced translation of the sense protein-coding mRNA. The data also confirmed translation of miRNA target mimics and lncRNAs that produce trans-acting or phased small-interfering RNA (tasiRNA/phasiRNAs). Mutational analyses of the positionally conserved sORF of TAS3a linked its translation with tasiRNA biogenesis. Altogether, this systematic analysis of ribosome-associated mRNAs and lncRNAs demonstrates that nutrient availability and translational regulation controls protein and small peptide-encoding mRNAs as well as a diverse cadre of regulatory RNAs.


Utilization of low-volume hospitals for total hip replacement.
To identify factors associated with utilization of low-volume hospitals for total hip replacement (THR) and to estimate differences in the distances that Medicare beneficiaries had to travel to reach low- or high-volume hospitals. We studied a population-based sample of 1,146 Medicare beneficiaries who underwent elective THR in 1995. Using multiple data sources including medical record review, Medicare claims data, 1990 Census data, and a patient survey, we examined factors independently associated with utilization of low-volume hospitals for elective THR. We estimated the magnitude of difference in distances for patients undergoing THR in low- and high-volume hospitals. We determined the distance between each patient's residence and the treating hospital using MapQuest. Rural residency, low income, and low educational attainment, as well as belief in the importance of convenient location in the choice of hospital, were associated with higher utilization of low-volume hospitals. Rural and suburban patients who went to low-volume hospitals traveled much less than patients operated upon in high-volume centers. Policies aimed at restricting THR to high-volume centers would differentially affect poor, less educated, and rural patients. Voluntary efforts to shift THR to high-volume centers should involve educating these patients and their referring physicians about differences in both short-term and longer-term outcomes between high- and low-volume centers.


Long-term outcome of 25 children and adolescents with severe aplastic anemia treated with antithymocyte globulin.
Severe aplastic anemia (SAA) is probably an immune-mediated disorder, and immunosuppressive therapy is recommended for patients with no available donor for bone marrow transplant. Between October 1984 and November 1987, 25 consecutive children and adolescents with SAA with no HLA-compatible marrow donor received equine antithymocyte globulin (ATG) (15 mg kg-1 day-1) for 10 days. The patients were evaluated 6 weeks, 6 months, and 12 months after starting ATG treatment. Thereafter, patients were evaluated yearly until July 1998. Median age was 10 years (range, 1.5-20 years), granulocyte counts on referral ranged from 0.032 to 1.4 x 10(9)/l (median 0.256 x 10(9)/l), and 12 patients had granulocyte counts <0.2 x 10(9)/l. At a median follow-up of 9.6 years (range, 8.6-11.8 years), 10 patients (40%) remained alive with good marrow function. No morphologic evidence of hematological clonal disorders has been observed, although two patients probably have acquired clonal chromosomal abnormalities (trisomy 8 and del(6)q21, respectively). Responses to ATG were observed between 6 weeks and 6 months from the start of treatment in 60% of evaluable patients. The response rate was not different in patients whose granulocyte count at diagnosis was <0.2 x 10(9)/l, or in those who were <10 years of age. This study supports the view that, when compared with supportive measures, ATG is an effective treatment for children or adolescents with SAA. Although these results are inferior to those reported for marrow transplantation or more intensive immunosuppressive regimens, these patients who responded to ATG are long-term survivors with stable peripheral blood counts and a low rate of relapse.


IgA pemphigus reacting exclusively to desmoglein 3.
IgA pemphigus is a rare disease which is classified into two major types: subcorneal pustular dermatosis (SPD) and intraepidermal neutrophilic IgA dermatosis (IEN). The autoantigen of the SPD type was identified as desmocollin1 (Dsc1), while the antigen of the IEN type is still unknown. We report a case of IgA pemphigus possessing antibodies exclusively against desmoglein3 (Dsg3). The patient also had ulcerative colitis, clinically severe oral mucosal lesions with pustule formation, and the "sunflower-like" pustular skin lesions. Histopathology showed intraepidermal bullae with numerous neutrophils. Direct immunofluorescence showed IgA deposits in the keratinocyte cell surfaces only in the lower epidermis. Indirect immunofluorescence using normal human skin as a substrate showed circulating IgA anti-cell surface autoantibodies at a titer of 1:160. cDNA transfection test for Dsc1-3 was negative. IgA antibodies to Dsg3, but not Dsg1, were detected with IgA-ELISA for Dsgs. Addition of diaphenylsulfone (DDS) to prednisolone and cyclosporine A significantly improved the oral and skin lesions. There have been rare cases showing IgA antibodies to either Dsg1 or Dsg3. These cases can be designated as IgA pemphigus foliaceus or IgA pemphigus vulgaris, respectively, according to the autoantigens. We report here a case of IgA pemphigus vulgaris complicated by ulcerative colitis.


Concentration-response relationships for salicylate-induced ototoxicity in normal volunteers.
1. Ototoxicity is a common and troublesome side-effect of high-dose aspirin treatment but there has been little previous study of the relationships between the degree of ototoxicity and the plasma concentrations of salicylate. 2. In order to investigate the relationships between aspirin dose, total and unbound plasma salicylate concentrations and ototoxicity, eight normal volunteers were dosed with aspirin 1.95, 3.25, 4.55 and 5.85 g day-1 for 1 week at each dose level, the doses being administered in random order and double-blind, 2 weeks apart. 3. Ototoxic effects measured were hearing loss in decibels (dB) over six frequencies and tinnitus intensity, estimated both by electronic matching and a fixed interval scale (FIS). Measurements were taken after steady-state concentrations of salicylate had been achieved. 4. Total and unbound plasma salicylate concentrations increased disproportionately with increasing daily doses of aspirin. The increase in the unbound salicylate was relatively greater since the percentage of salicylate unbound in plasma increased over the dose range investigated from a mean of 3.9% to 10.4%. 5. Hearing loss and tinnitus intensity increased progressively with the aspirin dosage and increasing concentrations of total and unbound plasma salicylate concentrations. These ototoxic symptoms were observed at lower concentrations of total salicylate than previously reported. 6. There was a linear relationship between hearing loss and unbound salicylate concentrations. 7. Further work is required to test the hypothesis that unbound plasma salicylate concentration is a better predictor of salicylate-induced ototoxicity than total plasma salicylate concentration.


Fluctuation of abundance and Lassa virus prevalence in Mastomys natalensis in Guinea, West Africa.
Based on empiric surveillance data, the incidence of human Lassa fever (LF) cases in Guinea and other West African countries has been reported to increase during the dry season compared to the rainy season. To investigate possible links with the ecology of the rodent reservoir of the virus, we conducted a 2-year longitudinal survey of Mastomys natalensis in a region of high human Lassa virus (LASV) seropositivity in Guinea. Standardized rodent trapping with similar trapping efforts between seasons was performed in three villages and 53.5% (601/1123) of the animals were identified as M. natalensis using morphometric and molecular criteria. Mean trapping success (TS) of M. natalensis was always higher inside houses than in proximal cultivations. In the dry season, mean TS increased 2-fold inside houses and decreased up to 10-fold outside (p < 0.0001), suggesting aggregation of rodents inside houses due to restricted food supply. 14.5% (80/553) of M. natalensis were tested positive for Lassa virus by reverse transcription-polymerase chain reaction (RT-PCR; range, 5%-30%) and prevalence of the virus was two to three times higher in rodents captured in the rainy season than in the dry season (p < 0.05). Inside houses, however, the LASV prevalence fluctuated nonsignificantly with season. These data suggest that in Guinea the risk of LASV transmission from rodents to humans is present both in the rainy and the dry season, reflected by the occurrence of LF cases throughout the year. In the dry season, however, the increased risk of humans encountering Mastomys and their excreta inside of houses may result in an increase of human Lassa fever cases.


The effect of sodium azide concentration on the recovery of enterococci from water.
The ability of Slanetz and Bartley medium to recover chlorine-stressed enterococci has been studied. Results showed that chlorine injury significantly affected the ability of Slanetz and Bartley medium to recover enterococci while lower concentrations of sodium azide in the same basal medium allowed their recovery. However, reducing the concentration of sodium azide considerably reduced the specificity making it unsuitable for use in the routine examination of water. A non-azide-containing medium, Enterolert(®)-DW appeared to be able to recover injured and non-injured enterococci with similar efficiency. The data presented here suggest that further work is required to improve the recovery of chlorine-injured enterococci by Slanetz and Bartley medium.


Impact of ferroelectric and superparaelectric nanoparticles on phase transitions and dynamics in nematic liquid crystals.
Results of broadband dielectric spectroscopy (BDS) studies of pure liquid crystalline (4-pentyloxy-4-biphenylcarbonitryle) 5OCB and its nanocolloids with BaTiO_{3} nanoparticles (NPs) under varying pressure and temperature are presented. The notable impact of NPs on phase transitions and dynamics was found. Particularly strong impact on pretransitional behavior was observed for relatively low concentrations of NPs, which can be related to the NPs-induced disorder. There are also notable differences between pressure and temperature paths of studies for nanocomposites, absent for the pure LC compound. For instance, tests focused on the translational orientational decoupling via the fractional Debye-Stokes-Einstein relation yielded S=0.71 and S=0.3 for the temperature and pressure paths, respectively: S=1 is for the complete coupling. The possible theoretical frame of observed phenomena is also proposed.


Fall History and Associated Factors Among Adults Living With HIV-1 in the Cape Winelands, South Africa: An Exploratory Investigation.
People with HIV-1 (PWH) exhibit a high fall incidence and increased fracture risk. As little is known about fall frequency and associated factors in PWH residing in lower-middle-income countries (LMIC), we investigated fall frequency, bone quality, and factors associated with fall history in a South African cohort. Fifty PWH without obvious predisposing factors for mobility impairments attending 2 public primary care clinics in the Western Cape region participated. Demographic, clinical, and physical performance data were collected. Falls were assessed retrospectively over 12 months. Mobility and balance were evaluated using a physical performance battery. Bone mineral density was screened using quantitative ultrasound (QUS). Associations between variables and falls grouping were analyzed using chi-square tests, t tests, and Mann-Whitney U tests, and effect sizes (ES) were calculated. Thirty-four percent of PWH (median age, 36.6 years) reported falling during the past year, and 41.2% of fallers reported multiple falls. Fallers had more mobility problems (P = .013), higher fear of falling (P = .007), higher fracture history (P = .003), worse balance performance (P < .001), higher proportions of detectable viral loads (P = .021), and poorer bone quality (P = .040). Differences were of medium to large ES. This exploratory study is the first to show that relatively young South African PWH without obvious predisposing factors for gait and balance impairments experience falls. The observed fall-associated factors warrant further research using larger samples and longitudinal designs to ascertain fall predictors within this population.


Polysome Profiling Links Translational Control to the Radioresponse of Glioblastoma Stem-like Cells.
Changes in polysome-bound mRNA (translatome) are correlated closely with changes in the proteome in cells. Therefore, to better understand the processes mediating the response of glioblastoma to ionizing radiation (IR), we used polysome profiling to define the IR-induced translatomes of a set of human glioblastoma stem-like cell (GSC) lines. Although cell line specificity accounted for the largest proportion of genes within each translatome, there were also genes that were common to the GSC lines. In particular, analyses of the IR-induced common translatome identified components of the DNA damage response, consistent with a role for the translational control of gene expression in cellular radioresponse. Moreover, translatome analyses suggested that IR enhanced cap-dependent translation processes, an effect corroborated by the finding of increased eIF4F-cap complex formation detected after irradiation in all GSC lines. Translatome analyses also predicted that Golgi function was affected by IR. Accordingly, Golgi dispersal was detected after irradiation of each of the GSC lines. In addition to the common responses seen, translatome analyses predicted cell line-specific changes in mitochondria, as substantiated by changes in mitochondrial mass and DNA content. Together, these results suggest that analysis of radiation-induced translatomes can provide new molecular insights concerning the radiation response of cancer cells. More specifically, they suggest that the translational control of gene expression may provide a source of molecular targets for glioblastoma radiosensitization. Cancer Res; 76(10); 3078-87. ©2016 AACR.


Pediatric gastrointestinal stromal tumors-a review of diagnostic modalities.
Gastrointestinal stromal tumors are exceedingly rare tumors in the pediatric population, as a result many clinicians either may never see this diagnosis or will encounter it only a few times throughout their careers. It is imperative in the pediatric population to follow appropriate steps to ensure a swift diagnosis and referral to specialized centers that are equipped with the multidisciplinary teams accustomed to treating rare diseases. This review aims to discuss the most recent data available on the diagnostic modalities utilized in cases of suspected Pediatric GIST.


[Reference Equations for the 6-Minute Walk Test in Healthy Individuals.]
The six-minute walk test (6MWT) has been broadly used in clinical settings. Several reference equations for prediction of the total distance walked during the test (6MWD) are available in literature. The present review aimed to critically discuss studies, published in Portuguese and English (LILACS, SCIELO, MEDLINE, PUBMED), which evaluated normal values and created reference equations for predicting 6MWD in healthy subjects, comparing them with the results that were recently obtained in Brazilian individuals. Age, sex, weight, height and body mass index were the main demographic and anthropometric features more often correlated with 6MWD. The equations derived from these characteristics were able to explain between 25 and 66% of the total variability in the 6MWD. Unfortunately, the foreign equations were not applicable to the Brazilian population. Even when the 6MWT was performed following strict standardization, the difference in 6MWT performance between foreign and Brazilian individuals remains, indicating the necessity of providing specific reference equations for each population and/or ethnic group. Accordingly, these equations developed in Brazil are probably the most appropriate for interpreting 6MWT performance in Brazilian patients with chronic diseases affecting the exercise capacity. Future studies are necessary with larger sample sizes (e.g. multicentric ones) and randomized design for the reference values of the 6MWD to be considered reliable.


[Correlations between placental morphological and functional parameters and neonatal status after normal full-term, prolonged pregnancy, and truly protracted pregnancy].
To solve the problem of differentiating protracted pregnancy, a special morphometric study was undertaken to examine three placental groups: 1) after normal full-term pregnancy (n = 35); 2) after prolonged pregnancy (n = 40); 3) after truly protracted pregnancy with partial or complete Clifford's syndrome (n = 30). A semiquantitative score (14 most important placental indicators), placenta and birth weights, Apgar scores, the infant's weight and height at the end of the first year of life were used so as to estimate the mild, moderate, and severe degree at 0.5, 1.5, and 3 points, respectively. One hundred and twenty-six possible pairs of structural and functional parameters were made up in all the groups. Pearson's correlation coefficient (r > 0.4) was applied to graphically display the pairs. Normal full-term pregnancy was characterized by few positive correlations mainly between the terminal villi and weight-height indices without associations with the Apgar scale. Prolonged pregnancy with the multiple placental tissue structural parameters along with organometric indices being involved was intermediate. The total scores permitted grades 1 and 2 chronic placental insufficiency (CPI) to be diagnosed in 27.5 and 2.5%, respectively. The severest form was truly protracted pregnancy that was distinguished by the maximum positive and negative correlations between all the parameters and the highest rate of diagnosis of CPI of grades 1 (45%) and 2 (15%). The findings strongly suggest that it is essential to identify prolonged and truly protracted pregnancy as important risk factors, by following up the infants for a year.


Mechanisms of auxin signaling.
The plant hormone auxin triggers complex growth and developmental processes. Its underlying molecular mechanism of action facilitates rapid switching between transcriptional repression and gene activation through the auxin-dependent degradation of transcriptional repressors. The nuclear auxin signaling pathway consists of a small number of core components. However, in most plants each component is represented by a large gene family. The modular construction of the pathway can thus produce diverse transcriptional outputs depending on the cellular and environmental context. Here, and in the accompanying poster, we outline the current model for TIR1/AFB-dependent auxin signaling with an emphasis on recent studies.


Nanoscale mapping of ion diffusion in a lithium-ion battery cathode.
The movement of lithium ions into and out of electrodes is central to the operation of lithium-ion batteries. Although this process has been extensively studied at the device level, it remains insufficiently characterized at the nanoscale level of grain clusters, single grains and defects. Here, we probe the spatial variation of lithium-ion diffusion times in the battery-cathode material LiCoO(2) at a resolution of ∼100 nm by using an atomic force microscope to both redistribute lithium ions and measure the resulting cathode deformation. The relationship between diffusion and single grains and grain boundaries is observed, revealing that the diffusion coefficient increases for certain grain orientations and single-grain boundaries. This knowledge provides feedback to improve understanding of the nanoscale mechanisms underpinning lithium-ion battery operation.


Are 20-hydroxyecdysone and related genes potential biomarkers of sublethal exposure to lipid-altering contaminants?
In Daphnia magna, 20-hydroecdysone (20E) is the main molting hormone and its metabolism is of interest to identify new biomarkers of exposure to contaminants. The present study aimed to (i) assess baseline levels of 20E and transcription levels of four related-genes (shade, neverland, ultraspiracle, and ecdysteroid receptor); and (ii) evaluate effects in D. magna after 21 days of exposure to fenarimol (anti-ecdysteroid) and a mixture of gemfibrozil and clofibric acid (lipid-lowering drugs) at sublethal concentrations. Endpoints included transcription of the target genes and quantification of 20E, mortality, and reproduction of daphnids. Baseline results showed that average responses were relatively similar and did not vary more than 2-fold. However, intra-day variation was generally high and could be explained by sampling individuals with slightly different stages of their development. Exposure tests indicated a significant decrease in daphnid reproduction following chronic exposure to a concentration of 565 μg/L of fenarimol. However, no difference was observed between the control and exposed groups for any of the investigated genes, nor for the levels of 20E after 21 days of exposure. Following exposition to gemfibrozil and clofibric acid at 1 μg/L, no changes were observed for the measured parameters. These results suggest that changes in transcription levels of the target genes and concentrations of 20E may not be sensitive endpoints that can be used as biomarkers of sublethal exposure to the target compounds in D. magna. Measuring multiple time points instead of a single measure as well as additional molecular endpoints obtained from transcriptomic and metabolomic studies could afford more insights on the changes occurring in exposed daphnids to lipid-altering compounds and identify efficient biomarkers of sublethal exposure.


Prioritization approaches in the development of health practice guidelines: a systematic review.
Given the considerable efforts and resources required to develop practice guidelines, developers need to prioritize what topics and questions to address. This study aims to identify and describe prioritization approaches in the development of clinical, public health, or health systems guidelines. We searched Medline and CINAHL electronic databases in addition to Google Scholar. We included papers describing prioritization approaches in sufficient detail allowing for reproducibility. We synthesized findings in a semi-quantitative way. We followed an iterative process to develop a common framework of prioritization criteria that captures all of the criteria reported by each included study. Our search captured 33,339 unique citations out of which we identified 10 papers reporting prioritization approaches for guideline development. All of the identified approaches focused on prioritizing guideline topics but none on prioritizing recommendation questions or outcomes. The two most frequently reported steps of the development process for these approaches were reviewing the grey literature (9 out of 10, 90%) and engaging various stakeholders (9 out of 10, 90%). We derived a common framework of 20 prioritization criteria that can be used when prioritizing guideline topics. The most frequently reported criteria were the health burden of disease which was included in all of the approaches, practice variation (8 out of 10, 80%), and impact on health outcomes (7 out of 10, 70%). Two of the identified approaches stood out as being comprehensive and detailed. We described 10 prioritization approaches in the development of health practice guidelines. There is a need to assess the effectiveness, efficiency and transparency of the identified approaches and to develop standardized and validated priority setting tools.


Return to Level I Sports After Anterior Cruciate Ligament Reconstruction: Evaluation of Age, Sex, and Readiness to Return Criteria.
Return to sport (RTS) after anterior cruciate ligament (ACL) reconstruction is a topic of current interest. The high reinjury rates reported in younger athletes may be due in part to an early RTS. To determine the proportion of athletes who return to level I (jumping, hard pivoting, and cutting) sport at less than 1 year after ACL reconstruction and compare return rates by age and sex. A secondary purpose was to examine whether RTS is associated with some commonly used outcome criteria. A cohort of 1440 athletes (992 males, 448 females) satisfied criteria for inclusion in this study (primary ACL reconstruction, normal contralateral knee, no additional surgery within the first year after the reconstruction, and participation in level I sport on a weekly basis prior to injury). The proportion of athletes in this sample who had resumed level I sport by 12 months after surgery was recorded, along with measurements of knee function (single-legged hop symmetry), laxity (KT-1000 arthrometer), and self-reported outcomes (International Knee Documentation Committee [IKDC] subjective score). Limb symmetry index scores of 90 or higher, side-to-side difference in anterior knee laxity of 2 mm or less, and IKDC scores of 95 or higher were considered indicators of satisfactory recovery from surgery. Rates of return to level I sport were significantly higher for athletes aged 25 years and younger (48% return rate) compared with older athletes (26-35 years, 32% return rate [P < .0001]; ≥36 years, 19% return rate [P < .001]). Male athletes who were in the ≤25-year and 26- to 35-year age brackets had significantly higher return rates than female athletes (52% vs 39% and 37% vs 18%, respectively), whereas no sex-based differences in RTS were seen after 36 years of age. A significantly higher proportion of younger patients met the selected indicators of a satisfactory recovery. Regardless of age, athletes had twice the odds of RTS if they had limb symmetry index scores of 90 or higher (P ≤ .0001) and 3 times the odds of RTS if they had IKDC scores of 95 or higher (P < .0001). Almost half of younger athletes (aged ≤25 years) in the current cohort returned to level I sports within 12 months after ACL reconstruction. Younger athletes were also more likely to meet criteria that indicate a satisfactory functional recovery and that have been used to indicate readiness to RTS. There may be a role for adjusting thresholds for these criteria based on age.


[Neurometabolic effects and antihypoxic activity of a vitamin-coenzyme complex, including thiamine pyrophosphate, lipoate, 4-phosphopantothenate, nicotinate and flavinadenine mononucleotide].
Administration of the vitamin-coenzyme complex containing thiamin pyrophosphate:lipoate:4-phosphopantothenate:nicotinate:flavinad enine mononucleotide (0.2:3:5:8:1) led to increased utilization of alpha-ketoglutarate in nervous tissue via alpha-ketoglutarate dehydrogenase and GABA-transaminase pathways. In these systems content of glutamate and GABA was decreased, suggesting their elevated consumption as energy-providing substrates. If 4-phosphopantothenate was substituted by CoASH and nicotinate--by NAD+, increased decarboxylation of 5(-14)C-alpha-ketoglutarate, 1(-14)C-GABA and 4(-14)C-Asp was detected in brain homogenates in vitro. The neurometabolic effects were accompanied by pronounced antihypoxic effects as shown by the test "hypoxy of closed space".


Photonic generation of ultra-wideband signals by direct current modulation on SOA section of an SOA-integrated SGDBR laser.
Photonic ultra-wideband (UWB) pulses are generated by direct current modulation of a semiconductor optical amplifier (SOA) section of an SOA-integrated sampled grating distributed Bragg reflector (SGDBR) laser. Modulation responses of the SOA section of the laser are first simulated with a microwave equivalent circuit model. Simulated results show a resonance behavior indicating the possibility to generate UWB signals with complex shapes in the time domain. The UWB pulse generation is then experimentally demonstrated for different selected wavelength channels with an SOA-integrated SGDBR laser.


Targeting Ca2+-dependent pathways to promote corneal epithelial wound healing induced by CISD2 deficiency.
Chronic epithelial defects of the cornea, which are usually associated with severe dry eye disease, diabetes mellitus, chemical injuries or neurotrophic keratitis, as well as aging, are an unmet clinical need. CDGSH Iron Sulfur Domain 2 (CISD2) is the causative gene for Wolfram syndrome 2 (WFS2; MIM 604928). CISD2 protein is significantly decreased in the corneal epithelium of patients with various corneal epithelial diseases. Here we summarize the most updated publications and discuss the central role of CISD2 in corneal repair, as well as providing new results describing how targeting Ca2+-dependent pathways can improve corneal epithelial regeneration. This review mainly focuses on the following topics. Firstly, an overview of the cornea and of corneal epithelial wound healing. The key players involved in this process, such as Ca2+, various growth factors/cytokines, extracellular matrix remodeling, focal adhesions and proteinases, are briefly discussed. Secondly, it is well known that CISD2 plays an essential role in corneal epithelial regeneration via the maintenance of intracellular Ca2+ homeostasis. CISD2 deficiency dysregulates cytosolic Ca2+, impairs cell proliferation and migration, decreases mitochondrial function and increases oxidative stress. As a consequence, these abnormalities bring about poor epithelial wound healing and this, in turn, will lead to persistent corneal regeneration and limbal progenitor cell exhaustion. Thirdly, CISD2 deficiency induces three distinct Ca2+-dependent pathways, namely the calcineurin, CaMKII and PKCα signaling pathways. Intriguingly, inhibition of each of the Ca2+-dependent pathways seems to reverse cytosolic Ca2+ dysregulation and restore cell migration during corneal wound healing. Notably, cyclosporin, an inhibitor of calcineurin, appears to have a dual effect on both inflammatory and corneal epithelial cells. Finally, corneal transcriptomic analyses have revealed that there are six major functional groupings of differential expression genes when CISD2 deficiency is present: (1) inflammation and cell death; (2) cell proliferation, migration and differentiation; (3) cell adhesion, junction and interaction; (4) Ca2+ homeostasis; (5) wound healing and extracellular matrix; and (6) oxidative stress and aging. This review highlights the importance of CISD2 in corneal epithelial regeneration and identifies the potential of repurposing venerable FDA-approved drugs that target Ca2+-dependent pathways for new uses, namely treating chronic epithelial defects of the cornea.


Modeling the effect of random roughness on synthetic aperture sonar image statistics.
A model has been developed to predict the effect of random seafloor roughness on synthetic aperture sonar (SAS) image statistics, based on the composite roughness approximation-a physical scattering model. The continuous variation in scattering strength produced by a random slope field is treated as an intensity scaling on the image speckle produced by the coherent SAS imaging process. Changes in image statistics caused by roughness are quantified in terms of the scintillation index (SI). Factors influencing the SI include the seafloor slope variance, geo-acoustic properties of the seafloor, the probability density function describing the speckle, and the signal-to-noise ratio. Example model-data comparisons are shown for SAS images taken at three different sites using three different high-frequency SAS systems. Agreement between the modeled and measured SI show that it is possible to link range-dependent image statistics to measurable geo-acoustic properties, providing the foundation necessary for solving problems related to the detection of targets using high-frequency imaging sonars, including performance prediction or adaptation of automated detection algorithms. Additionally, this work illustrates the possible use of SAS systems for remote sensing of roughness parameters such as root mean square slope or height.


Hawthorn (Crataegus pinnatifida Bunge) leave flavonoids attenuate atherosclerosis development in apoE knock-out mice.
Hawthorn (Crataegus pinnatifida Bunge) leave have been used to treat cardiovascular diseases in China and Europe. Hawthorn leave flavonoids (HLF) are the main part of extraction. Whether hawthorn leave flavonoids could attenuate the development of atherosclerosis and the possible mechanism remain unknown. High-fat diet (HFD) mixed with HLF at concentrations of 5mg/kg and 20mg/kg were administered to apolipoprotein E (apoE) knock out mice. 16 weeks later, mouse serum was collected to determine the lipid profile while the mouse aorta dissected was prepared to measure the lesion area. Hepatic mRNA of genes involved in lipid metabolism were determined. Peritoneal macrophages were collected to study the impact of HLF on cholesterol efflux, formation of foam cell and the expression of ATP binding cassette transporter A1 (ABCA1). Besides, in vivo reverse cholesterol transport (RCT) was conducted. HLF attenuated the development of atherosclerosis that the mean atherosclerotic lesion area in en face aortas was reduced by 23.1% (P<0.05). In mice fed with 20mg/kg HLF, Total cholesterol (TC) level was decreased by 18.6% and very low density lipoprotein cholesterol plus low density lipoprotein cholesterol (VLDLc+LDLc) level were decreased by 23.1% whereas high density lipoprotein cholesterol (HDLc) and triglyceride (TG) levels were similar compared to that of the control group. Peroxisome proliferator activated receptor alpha (PPARα) mRNA was increased by 31.2% (P<0.05) and 60.9% (P<0.05) in mice fed with 5mg/kg and 20mg/kg HLF respectively. Sterol regulatory element binding protein-1c (SREBP-1c) was decreased by 59.3% in the group of 20mg/kg. Carnitine palmitoyl transferase 1 (CPT-1) mRNA level of 20mg/kg group was induced 66.7% (P<0.05). Superoxide dismutase 1 and 2 (SOD1 and SOD2) mRNA were induced 25.4% (P<0.05) and 71.4% (P<0.05) while induced by 36.3% (P<0.05) and 73.2% (P<0.05) in group of 20mg/kg. Glutathione peroxidase 3 (Gpx3) mRNA in the group of 20mg/kg was induced by 96.7% (P<0.05). Hepatic hydroxymethylglutaryl CoA reductase (HMG-CoAR) expression was as same level as the control group while LDL receptor (LDLR) mRNA and protein were induced by 84.2% (P<0.05) and 98.8% (P<0.05) in group of 20mg/kg. HLF inhibit the formation of foam cell by 27.9% (P<0.05) in the dosage of 25μg/ml, and 33.3% (P<0.05) in the dosage of 50μg/ml. HLF increased the reverse cholesterol transport (RCT) in vivo. Hawthorn leave flavonoids can slow down the development of atherosclerosis in apoE knockout mice via induced expression of genes involved in antioxidant activities, inhibition of the foam cell formation and promotion of RCT in vivo, which implies the potential use in the prevention of atherosclerosis.


Oral retinoids and plasma lipids.
Retinoids and rexinoids are prescribed for conditions ranging from acne vulgaris to hyperkeratosis to cutaneous T cell lymphoma. Dyslipidemia is a frequent consequence of the use of these drugs, with more than one-third of patients manifesting aberrations in triglyceride (TG) levels. The efficacy of retinoic acid derivatives is linked to their influence on lipid metabolism in the skin, which can impair systemic lipid trafficking and metabolism in some patients. Thus, baseline screening for preexisting dyslipidemia and regular follow-up lipid panels are mandated, especially when powerful agents such as bexarotene are used. Dietary modification, increased physical activity, and weight management are the cornerstones of initial management for mild hypertriglyceridemia, which is a contributor to cardiovascular risk. More severe impairments (fasting TG > 500 mg/dL) warrant pharmacologic interventions early on to reduce the risk of pancreatitis. Retinoic acid derivative action, lipid metabolism, and treatment of incident dyslipidemias are reviewed to empower prescribers in management of adverse lipid effects.


Liver transplantation-associated de novo hepatitis B virus infection: application of molecular evolutionary analysis.
De novo hepatitis B virus (HBV) infection after liver transplantation has recently been reported to be associated with donors without serum hepatitis B surface antigen (HbsAg) but with hepatitis B core antigen (anti-HBc). We elucidate the source of de novo HBV infection after liver transplantation by molecular evolutionary analysis. The serum sample was obtained from a recipient who underwent living related liver transplantation. He was negative for all HBV-related serum markers before the transplantation. The recipient became seropositive for HBsAg at 6 months after transplantation. The liver tissue was obtained from a donor who was seronegative for HBsAg, but positive for anti-HBs and anti-HBc. HBV DNA was detected from the serum and liver tissue in a recipient and donor, respectively. A total of 5 clones each of small-S gene of HBV from the donor and recipient were sequenced. A phylogenetic tree analysis based on small-S gene revealed that all isolates derived from the recipient and donor were clustered together within a close range of evolutionary distances. These results indicated that HBV was transmitted by the liver graft from the donor. Molecular evolutionary analysis can be adopted for the study of the transmission route of viral infection via organ transplantation.


Reversible solid state redox of an octacyanometallate-bridged coordination polymer by electrochemical ion insertion/extraction.
Coordination polymers have significant potential for new functionality paradigms due to the intrinsic tunability of both their electronic and structural properties. In particular, octacyanometallate-bridged coordination polymers have the extended structural and magnetic diversity to achieve novel functionalities. We demonstrate that [Mn(H2O)][Mn(HCOO)(2/3)(H2O)(2/3)](3/4)[Mo(CN)8]·H2O can exhibit electrochemical alkali-ion insertion/extraction with high durability. The high durability is explained by the small lattice change of less than 1% during the reaction, as evidenced by ex situ X-ray diffraction analysis. The ex situ X-ray absorption spectroscopy revealed reversible redox of the octacyanometallate. Furthermore, the solid state redox of the paramagnetic [Mo(V)(CN)8](3-)/diamagnetic[Mo(IV)(CN)8](4-) couple realizes magnetic switching.


Sp7 and Runx2 molecular complex synergistically regulate expression of target genes.
Runx2 and Sp7 transcription factors are essential for skeletogenesis. Targeted deletion of either gene results in failure of osteoblast differentiation and bone formation. Loss of bone-matrix gene expression is surprisingly similar in Sp7 and Runx2 null mice. The molecular mechanisms responsible for similar transcriptional regulation of target genes remain largely unknown. Here, we demonstrate that Runx2 and Sp7 interact physically and functionally. Both proteins are co-expressed in osteoblastic cells. We first characterized a panel of Sp7 antibodies and demonstrate that majority of the published antibodies do not recognize Sp7 protein. Co-immunoprecipitation studies revealed that endogenous Runx2 protein physically interacts with Sp7 protein. We identified that runt homology domain (RHD) of Runx2 protein is involved in physical association with Sp7. Functional consequences of Runx2-Sp7 physical interaction was then assessed by promoter-reporter assays. We selected promoters of osteocalcin (OC), a marker of mature osteoblast and fibroblast growth factor 3 (FGF3), a signaling molecule that determine the fate of embryonic ecto-mesenchyme. Runx2 and Sp7 stimulate OC-promoter activity by 3-folds in epithelial cells. However, when both proteins were co-expressed, a dose-dependent synergistic activation of 22-folds was noted. Similar pattern of synergistic activation of OC-promoter was noted in mesenchymal cell. FGF3 promoter was activated by 25 - and 30-folds with Runx2 and Sp7 respectively. Again a dose-dependent synergistic activation of 130-folds was evident when Runx2 and Sp7 were co-expressed in epithelial cells. Synergistic activation of FGF3 promoter was also noted in mesenchymal cells. Together, our data demonstrated that Runx2-Sp7 molecular complex functionally cooperate for maximal induction of cell-phenotype-restricted genes.


Chromosome Engineering of Human Colon-Derived Organoids to Develop a Model of Traditional Serrated Adenoma.
Traditional serrated adenomas (TSAs) are rare colorectal polyps with unique histologic features. Fusions in R-spondin genes have been found in TSAs, but it is not clear whether these are sufficient for TSA development, due to the lack of a chromosome engineering platform for human tissues. We studied the effects of fusions in R-spondin genes and other genetic alterations found in TSA using CRISPR-Cas9-mediated chromosome and genetic modification of human colonic organoids. We introduced chromosome rearrangements that involve R-spondin genes into human colonic organoids, with or without disruption of TP53, using CRISPR-Cas9 (chromosome-engineered organoids). We then knocked a mutation into BRAF encoding the V600E substitution and overexpressed the GREM1 transgene; the organoids were transplanted into colons of NOG mice and growth of xenograft tumors was measured. Colon tissues were collected and analyzed by immunohistochemistry or in situ hybridization. We also established 2 patient-derived TSA organoid lines and characterized their genetic features and phenotypes. We inserted a bicistronic cassette expressing a dimerizer-inducible suicide gene and fluorescent marker downstream of the LGR5 gene in the chromosome-engineered organoids; addition of the dimerizer eradicates LGR5+ cells. Some tumor-bearing mice were given intraperitoneal injections of the dimerizer to remove LGR5-expressing cells. Chromosome engineering of organoids required disruption of TP53 or culture in medium containing IGF1 and FGF2. In colons of mice, organoids that expressed BRAFV600E and fusions in R-spondin genes formed flat serrated lesions. Patient-derived TSA organoids grew independent of exogenous R-spondin, and 1 line grew independent of Noggin. Organoids that overexpressed GREM1, in addition to BRAFV600E and fusions in R-spondin genes, formed polypoid tumors in mice that had histologic features similar to TSAs. Xenograft tumors persisted after loss of LGR5-expressing cells. We demonstrated efficient chromosomal engineering of human normal colon organoids. We introduced genetic and chromosome alterations into human colon organoids found in human TSAs; tumors grown from these organoids in mice had histopathology features of TSAs. This model might be used to study progression of human colorectal tumors with RSPO fusion gene and GREM1 overexpression.


Relation of coronary calcium scoring with cardiovascular events in patients with diabetes: The CLARIFY Registry.
Coronary artery calcium (CAC) scoring is not routinely performed in patients with diabetes based on an existing class I indication for statin therapy in these patients. However, CAC scoring may improve risk classification and prediction of atherosclerotic cardiovascular disease (ASCVD) events beyond risk scores in asymptomatic individuals with prediabetes and diabetes, warranting CAC assessment in this population. The routine availability through provision of no-charge CAC as an alternative to routine probabilistic risk scores may improve utilization of preventive therapies especially in traditionally underserved populations. Prospective observational study in a large health system offering no-charge CAC scoring for primary prevention risk prediction with available glycosylated hemoglobin (HbA1c) measurements between June 2015 and March 2019 were divided according to no diabetes (HbA1c <5.7 %), prediabetes (HbA1c 5.7 %-6.4 %), or diabetes (HbA1c ≥ 6.5 % or charted history) and followed for major adverse cardiovascular events [myocardial infarction, stroke, death (MACE) or coronary revascularization]. Patient characteristics, health history, laboratory data, and statin prescription rates were measured at baseline and at one year after CAC scoring. A total of 12,194 subjects with available HbA1c underwent CAC scoring during the study period (6462 diabetes, 2062 prediabetes, and 3670 without diabetes). At a median follow-up of 1.2 years, there were 458 MACE events (71 patients without diabetes, 66 patients with prediabetes, and 321 patients with diabetes). Among patients with diabetes or prediabetes, increased CAC was associated with MACE (HR 1.38 [1.26-1.51], p < 0.001) and MACE or revascularization (HR 1.70 [1.57-1.85], p < 0.001). In patients with diabetes, CAC category was associated with greater statin initiation (89.6 % for CAC≥400 vs 60.1 % for CAC = 0, p < 0.001) and high intensity statin initiation (42.2 % for CAC≥400 vs 16.8 % for CAC = 0, p < 0.001) at one year post CAC scoring. Patients with diabetes had greater reductions in systolic blood pressure, LDL-C, total cholesterol, and triglycerides from baseline with a CAC ≥400 compared to a lower CAC category (p = 0.007). CAC burden is associated with ASCVD risk in patients with diabetes. CAC scoring increases statin prescriptions and reduces ASCVD risk in patients with diabetes, potentially warranting routine CAC assessment in this population.


The European Forest Condition Monitor: Using Remotely Sensed Forest Greenness to Identify Hot Spots of Forest Decline.
Forest decline, in course of climate change, has become a frequently observed phenomenon. Much of the observed decline has been associated with an increasing frequency of climate change induced hotter droughts while decline induced by flooding, late-frost, and storms also play an important role. As a consequence, tree mortality rates have increased across the globe. Despite numerous studies that have assessed forest decline and predisposing factors for tree mortality, we still lack an in-depth understanding of (I) underlying eco-physiological mechanisms, (II) the influence of varying environmental conditions related to soil, competition, and micro-climate, and (III) species-specific strategies to cope with prolonged environmental stress. To deepen our knowledge within this context, studying tree performance within larger networks seems a promising research avenue. Ideally such networks are already established during the actual period of environmental stress. One approach for identifying stressed forests suitable for such monitoring networks is to assess measures related to tree vitality in near real-time across large regions by means of satellite-borne remote sensing. Within this context, we introduce the European Forest Condition monitor (EFCM)-a remote-sensing based, freely available, interactive web information tool. The EFCM depicts forest greenness (as approximated using NDVI from MODIS at a spatial resolution of roughly 5.3 hectares) for the pixel-specific growing season across Europe and consequently allows for guiding research within the context of concurrent forest performance. To allow for inter-temporal comparability and account for pixel-specific features, all observations are set in relation to normalized difference vegetation index (NDVI) records over the monitoring period beginning in 2001. The EFCM provides both a quantile-based and a proportion-based product, thereby allowing for both relative and absolute comparison of forest greenness over the observational record. Based on six specific examples related to spring phenology, drought, late-frost, tree die-back on water-logged soils, an ice storm, and windthrow we exemplify how the EFCM may help identifying hotspots of extraordinary forest greenness. We discuss advantages and limitations when monitoring forest condition at large scales on the basis of moderate resolution remote sensing products to guide users toward an appropriate interpretation.


Novel promoter upstream of the human c-myc gene and regulation of c-myc expression in B-cell lymphomas.
A new promoter of the human c-myc gene called P0, with multiple RNA start sites, was mapped over 500 bases upstream of the two previously identified promoters, P1 and P2. Sequencing full-length cDNA clones of P0 RNAs revealed two open reading frames upstream of that for the P64c-myc protein. P0 RNA is located on polyribosomes and released by puromycin, indicating that it functions as an mRNA. In vitro translation of RNA synthesized from the cloned cDNAs predicts that P0 transcripts are translated into a novel 12.5-kilodalton protein corresponding to the first open reading frame. The regulation of P0 RNA was studied in the B-cell lymphoma cell line Manca, in which only the translocated c-myc allele lacking exon 1 was thought to be active. However, we found that P0 transcription and the DNase I-hypersensitive site associated with this promoter persist on the untranslocated allele, even though P1/P2 transcription as measured by a nuclear runoff assay was repressed. These results suggest that allelic exclusion of c-myc expression in this B-cell lymphoma is caused by a repression of transcription which is specific to the P1/P2 promoters. We previously reported a block to elongation of transcription near the 3' end of exon 1 in the wild-type c-myc gene, which results in an excess of exon 1 over exon 2 transcription (5a). In contrast, we found that in the Daudi B-cell lymphoma, which retains exon 1 in the active allele, equimolar transcription of exons 1 and 2 occurs. This result suggests a model for the activation of c-myc in B-cell lymphomas.


Learning a commonsense moral theory.
We introduce a computational framework for understanding the structure and dynamics of moral learning, with a focus on how people learn to trade off the interests and welfare of different individuals in their social groups and the larger society. We posit a minimal set of cognitive capacities that together can solve this learning problem: (1) an abstract and recursive utility calculus to quantitatively represent welfare trade-offs; (2) hierarchical Bayesian inference to understand the actions and judgments of others; and (3) meta-values for learning by value alignment both externally to the values of others and internally to make moral theories consistent with one's own attachments and feelings. Our model explains how children can build from sparse noisy observations of how a small set of individuals make moral decisions to a broad moral competence, able to support an infinite range of judgments and decisions that generalizes even to people they have never met and situations they have not been in or observed. It also provides insight into the causes and dynamics of moral change across time, including cases when moral change can be rapidly progressive, changing values significantly in just a few generations, and cases when it is likely to move more slowly.


[Fibromyalgia--an update].
Fibromyalgia is a common, female dominated condition, characterized by chronic widespread pain, fatigue, sleep disturbance, anxiety/depression and a number of other symptoms. The syndrome's aetiology and treatment is disputed. However, research over the last decade has provided substantial new information. This review is an update on fibromyalgia with respect to predictors, pathogenesis and treatment. On the basis of a number of important scientific publications, the most relevant theory is presented and discussed. Fibromyalgia may be described as a biopsychosocial disorder in which long lasting distress, myofascial pain, central sensitisation, sickness behaviour, anxiety/depression, neuroendocrine and autonomic dysfunction play central aetiological roles as elements in a vicious circle. Treatment should be focused on each of these elements and requires active patient participation. It is best given as a multidimensional, multidisciplinary rehabilitation programme with continuous support from the general practitioner. Even though the fibromyalgia riddle is not yet fully solved, research over the last decade has given valuable new information on pathogenesis and treatment. We should be able to offer our patients substantial help on the basis of present knowledge.


Factors Associated With Maintenance of an Improved Ejection Fraction: An Echocardiogram-Based Registry Study.
Background Heart failure with improved ejection fraction (EF) is increasingly recognized as a sizable and distinct entity. While the features associated with improvedEF have been explored and new guidelines have emerged, factors associated with sustaining an improved EF over time have not been defined. We aimed to assess factors associated with maintenance of an improved EF in a large real-world patient cohort. Methods and Results A total of 7070 participants with heart failure with improved EF and a subsequent echocardiogram performed after at least 9 months of follow-up were included in a retrospective cohort study conducted at the Cleveland Clinic in Cleveland, Ohio. Multiple logistic regression models, adjusted for demographics, comorbidities, and medications were built to identify characteristics and therapeutic interventions associated with maintaining an improved EF. Mean age (SD) was 64.9 (13.8) years, 62.7% were men, and 75.1% were White participants. White race and the use of angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, or angiotensin receptor-neprilysin inhibitors were associated with maintaining the EF at least 9 months after EF improvement. In contrast, male sex or having atrial fibrillation/flutter, coronary artery disease, history of myocardial infarction, presence of an implanted cardioverter-defibrillator, and use of loop diuretics were associated with a decline in EF after previously documented improvement. Conclusions Continued use of renin-angiotensin-aldosterone system inhibitors was associated with maintaining the EF beyond the initial improvement phase.


Circulating IL-6, IL-10, and TNF-alpha and IL-10/IL-6 and IL-10/TNF-alpha ratio profiles of polyparasitized individuals in rural and urban areas of gabon.
Malaria, blood-borne filarial worms and intestinal parasites are all endemic in Gabon. This geographical co-distribution leads to polyparasitism and, consequently, the possibility of immune-mediated interactions among different parasite species. Intestinal protozoa and helminths could modulate antimalarial immunity, for example, thereby potentially increasing or reducing susceptibility to malaria. The aim of the study was to compare the cytokine levels and cytokine ratios according to parasitic profiles of the population to determine the potential role of co-endemic parasites in the malaria susceptibility of populations. Blood and stool samples were collected during cross-sectional surveys in five provinces of Gabon. Parasitological diagnosis was performed to detect plasmodial parasites, Loa loa, Mansonella perstans, intestinal helminths (STHs) and protozoan parasites. Nested PCR was used to detect submicroscopic plasmodial infection in individuals with negative blood smears. A cytometric bead array was used to quantify interleukin (IL)-6, IL-10 and tumour necrosis factor (TNF)-α in the plasma of subjects with different parasitological profiles. Median IL-6 and IL-10 levels and the median IL-10/TNF-α ratio were all significantly higher among individuals with Plasmodium (P.) falciparum infection than among other participants (p<0.0001). The median TNF-α level and IL-10/IL-6 ratio were higher in subjects with STHs (p = 0.09) and P. falciparum-intestinal protozoa co-infection (p = 0.04), respectively. IL-6 (r = -0.37; P<0.01) and IL-10 (r = -0.37; P<0.01) levels and the IL-10/TNF-α ratio (r = -0.36; P<0.01) correlated negatively with age. Among children under five years old, the IL-10/TNF-α and IL-10/IL-6 ratios were higher in those with intestinal protozoan infections than in uninfected children. The IL-10/TNF-α ratio was also higher in children aged 5-15 years and in adults harbouring blood-borne filariae than in their control counterparts, whereas the IL-10/IL-6 ratio was lower in those aged 5-15 years with filariae and intestinal parasites but higher in adults with intestinal parasitic infections. Asymptomatic malaria is associated with a strong polarization towards a regulatory immune response, presenting high circulating levels of IL-10. P. falciparum/intestinal protozoa co-infections were associated with an enhanced IL-10 response. Immunity against malaria could differ according to age and carriage of other parasites. Helminths and intestinal protozoa can play a role in the high susceptibility to malaria currently observed in some areas of Gabon, but further investigations are necessary.


Bone marrow-derived cells and biophysical stimulation for talar osteochondral lesions: a randomized controlled study.
Osteochondral lesions of the talus (OLT) frequently occur after ankle sprains in young patients participating in sports activities. These injuries may lead to chronic pain, joint swelling, and finally osteoarthritis, therefore, surgical repair is frequently needed. A collagen scaffold seeded with bone marrow-derived cells (BMDCs) harvested from patient's iliac crest and implanted into the OLT through a single arthroscopic procedure has been recently proposed as an effective treatment option. Nevertheless, BMDCs, embedded in an inflammatory environment, tend to differentiate toward a fibroblast phenotype with a consequential loss of mechanical characteristics. Biophysical stimulation with pulsed electromagnetic fields (PEMFs) has been shown to promote anabolic chondrocyte activity, stimulate proteoglycan synthesis, and reduce the release of the most relevant pro-inflammatory cytokines. The aim of this randomized controlled trial was to evaluate the effects of PEMFs on clinical outcome in patients who underwent BMDCs transplantation for OLT. Thirty patients affected by grade III and IV Outerbridge OLT underwent BMDCs transplantation. After surgery, patients were randomly assigned to either experimental group (PEMFs 4 hours per day for 60 days starting within 3 days after operation) or control group. Clinical outcome was evaluated with (American Orthopaedic Foot and Ankle Society) AOFAS score, Visual Analog Scale (VAS), and Short Form-36 (SF-36). Significantly higher AOFAS score was recorded in the experimental group both at 6 or 12 months follow-up. At 60 days and 6 and 12 months follow-up, significant lower pain was observed in the experimental group. No significant difference was found in SF-36 between groups. A superior clinical outcome was found in the experimental group with more than 10 points higher AOFAS score at final follow-up. Biophysical stimulation started soon after surgery aided patient recovery leading to pain control and a better clinical outcome with these improvements lasting more than 1 year after surgery. Level II, prospective comparative study.


DNA-free CRISPR-Cas9 gene editing of wild tetraploid tomato Solanum peruvianum using protoplast regeneration.
Wild tomatoes (Solanum peruvianum) are important genomic resources for tomato research and breeding. Development of a foreign DNA-free clustered regularly interspaced short palindromic repeat (CRISPR)-Cas delivery system has potential to mitigate public concern about genetically modified organisms. Here, we established a DNA-free CRISPR-Cas9 genome editing system based on an optimized protoplast regeneration protocol of S. peruvianum, an important resource for tomato introgression breeding. We generated mutants for genes involved in small interfering RNAs biogenesis, RNA-DEPENDENT RNA POLYMERASE 6 (SpRDR6), and SUPPRESSOR OF GENE SILENCING 3 (SpSGS3); pathogen-related peptide precursors, PATHOGENESIS-RELATED PROTEIN-1 (SpPR-1) and PROSYSTEMIN (SpProSys); and fungal resistance (MILDEW RESISTANT LOCUS O, SpMlo1) using diploid or tetraploid protoplasts derived from in vitro-grown shoots. The ploidy level of these regenerants was not affected by PEG-Ca2+-mediated transfection, CRISPR reagents, or the target genes. By karyotyping and whole genome sequencing analysis, we confirmed that CRISPR-Cas9 editing did not introduce chromosomal changes or unintended genome editing sites. All mutated genes in both diploid and tetraploid regenerants were heritable in the next generation. spsgs3 null T0 regenerants and sprdr6 null T1 progeny had wiry, sterile phenotypes in both diploid and tetraploid lines. The sterility of the spsgs3 null mutant was partially rescued, and fruits were obtained by grafting to wild-type (WT) stock and pollination with WT pollen. The resulting seeds contained the mutated alleles. Tomato yellow leaf curl virus proliferated at higher levels in spsgs3 and sprdr6 mutants than in the WT. Therefore, this protoplast regeneration technique should greatly facilitate tomato polyploidization and enable the use of CRISPR-Cas for S. peruvianum domestication and tomato breeding.


Efficacy and safety of canakinumab in patients with Still's disease: exposure-response analysis of pooled systemic juvenile idiopathic arthritis data by age groups.
To describe the efficacy, safety, and exposure-response relationship of canakinumab in a subgroup of patients with systemic juvenile idiopathic arthritis (SJIA) aged ≥16 years, representative of adult-onset Still's disease (AOSD) patients, and to compare this subgroup with those of children and young adolescents with SJIA by pooling clinical data collected during the development programme of canakinumab. Safety and efficacy data on canakinumab-treated patients were pooled from 4 SJIA studies (NCT00426218, NCT00886769, NCT00889863, and NCT00891046). In the majority of patients, canakinumab was administered at 4 mg/kg every 4 weeks. Efficacy parameters (adapted American College of Rheumatology [aACR] paediatric and juvenile idiopathic arthritis [JIA] ACR responses), quality of life, C-reactive protein levels, safety, and exposure-response relationship were assessed over 12 weeks in 3 age groups (children 2-<12, young adolescents 12-<16 and older adolescents and young adults ≥16 years). Efficacy outcomes were analysed in 216 children, 56 young adolescents and 29 older adolescents and young adults. Efficacy parameters across 3 age groups were largely comparable. At Day 15, at least 50% of patients from each age group exhibited aACR ≥70 and ACR responses. The safety profile of canakinumab was similar across age groups. One death was reported. Pooled analyses from SJIA studies indicate that older adolescents and young adults SJIA patients show similar efficacy, safety, and exposure-response relationship on a weight-based dosing regimen as observed in children and adolescent SJIA patients. These analyses suggest that canakinumab may be an effective therapy in young adults with Still's disease.


Regulation of muscarinic acetylcholine receptor number in cultured neuronal cells by chronic membrane depolarization.
The effect of chronic membrane depolarization on the regulation of muscarinic acetylcholine receptor (mAChR) number was studied in neuroblastoma cells (clone N1E-115). Receptor number was determined by a filter binding assay using 3H-quinuclidinyl benzilate (QNB) in membrane and crude cellular homogenates. Incubation with 50 microM veratridine (VTN), an activator of voltage-sensitive Na+ channels, induced a 50-200% increase in mAChR number at 24 hr, which was inhibited 80% by TTX. Scatchard analysis showed that affinity of the mAChR for 3H-QNB was not affected by VTN. Upon withdrawal of VTN, mAChR number returned to control levels within 20 hr. Chronic membrane depolarization caused by incubation in medium containing 60 mM K+ induced a TTX-insensitive 50% increase in mAChR number at 24 hr. AChE activity was unaffected by chronic membrane depolarization. The VTN-induced increase in mAChR number was not blocked by coincubation with cycloheximide or tunicamycin, both inhibitors of de novo mAChR synthesis. The rate of mAChR degradation was reduced in the presence of 50 microM VTN, with the apparent half-life increased from approximately 18 hr (control) to approximately 40 hr (VTN). Although treatment with either 1 mM 8Br-cAMP or 1 mM 8Br-GMP failed to increase mAChR number, treatment with either the inorganic Ca2+ channel blocker Co2+ (1 mM) or the organic Ca2+ channel antagonist D600 (10-100 microM) produced 40-80% increases in mAChR number. The combination of VTN and either D600 or Co2+ failed to induce a greater increase in mAChR number than incubation with VTN alone.(ABSTRACT TRUNCATED AT 250 WORDS)