Pralatrexate Sustainably Released from Polypeptide Thermogel Is Effective for Chondrogenic Differentiation of Mesenchymal Stem Cells.
Folic acid was reported to significantly improve chondrogenic differentiation of mesenchymal stem cells. In a similar mechanism of action, we investigated clinically approved antifolates by the U.S. Food and Drug Administration as chondrogenic-promoting compounds for tonsil-derived mesenchymal stem cells. A poly(ethylene glycol)-poly(l-alanine) thermogelling system was used as a three-dimensional cell culture matrix, where stem cells and antifolates could be incorporated simultaneously during a heat-induced in situ sol-to-gel transition. The antifolates could be supplied over several days by the sustained release of the drug from the thermogel. Initially, seven antifolates were prescreened based on cell viability and expression of a typical chondrogenic biomarker of type II collagen (COL II) at the mRNA level. Then, dapsone, pralatrexate, and trimethoprim were selected as candidate compounds in the second round screening, and detailed studies were carried out on the mRNA and protein expression of various chondrogenic biomarkers including COL II, SRY box transcription factor 9, and aggrecan. Three-dimensional cultures of stem cells in the thermogel in the absence of a chondrogenic promoter compound and in the presence of kartogenin (KGN) were performed as a negative control and positive control, respectively. The chondrogenic biomarkers were significantly increased in the selected antifolate-incorporating systems compared to the negative control system, without an increase in type I collagen (an osteogenic biomarker) expression. Pralatrexate was the best compound for inducing chondrogenic differentiation of the stem cells, even better than the positive control (KGN). Nuclear translocation of the core-binding factor β subunit (CBFβ) and enhanced nuclear runt-related transcription factor 1 (RUNX1) by antifolate treatment suggested that the chondrogenesis-enhancing mechanism is mediated by CBFβ and RUNX1. An in silico modeling study confirmed the mechanism by proving the high binding affinity of pralatrexate to a target protein of filamin A compared with other antifolate candidates. To conclude, pralatrexate was rediscovered as a lead compound, and the polypeptide thermogel incorporating pralatrexate and mesenchymal stem cells can be a very effective system in promoting chondrogenic differentiation of stem cells and might be used in injectable tissue engineering for cartilage repair.


Study of hepatitis E virus infection of genotype 1 and 3 in mice with humanised liver.
The hepatitis E virus (HEV) is responsible for approximately 20 million infections per year worldwide. Although most infected people can spontaneously clear an HEV infection, immune-compromised individuals may evolve towards chronicity. Chronic HEV infection can be cured using ribavirin, but viral isolates with low ribavirin sensitivity have recently been identified. Although some HEV isolates can be cultured in vitro, in vivo studies are essentially limited to primates and pigs. Since the use of these animals is hampered by financial, practical and/or ethical concerns, we evaluated if human liver chimeric mice could serve as an alternative. Humanised mice were inoculated with different HEV-containing preparations. Chronic HEV infection was observed after intrasplenic injection of cell culture-derived HEV, a filtered chimpanzee stool suspension and a patient-derived stool suspension. The viral load was significantly higher in the stool compared with the plasma. Overall, the viral titre in genotype 3-infected mice was lower than that in genotype 1-infected mice. Analysis of liver tissue of infected mice showed the presence of viral RNA and protein, and alterations in host gene expression. Intrasplenic injection of HEV-positive patient plasma and oral inoculation of filtered stool suspensions did not result in robust infection. Finally, we validated our model for the evaluation of novel antiviral compounds against HEV using ribavirin. Human liver chimeric mice can be infected with HEV of different genotypes. This small animal model will be a valuable tool for the in vivo study of HEV infection and the evaluation of novel antiviral molecules.


A study protocol for a Pilot Masked, Randomized Controlled Trial Evaluating Locally-applied Gentamicin versus Saline in Open Tibia Fractures (pGO-Tibia) in Dar es Salaam, Tanzania.
Open tibia fractures are a major source of disability in low- and middle-income countries (LMICs) due to the high incidence of complications, particularly infection and chronic osteomyelitis. One proposed adjunctive measure to reduce infection is prophylactic local antibiotic delivery, which can achieve much higher concentrations at the surgical site than can safely be achieved with systemic administration. Animal studies and retrospective clinical studies support the use of gentamicin for this purpose, but no high-quality clinical trials have been conducted to date in high- or low-income settings. We describe a protocol for a pilot study conducted in Dar es Salaam, Tanzania, to assess the feasibility of a single-center masked randomized controlled trial to compare the efficacy of locally applied gentamicin to placebo for the prevention of fracture-related infection in open tibial shaft fractures. The results of this study will inform the design and feasibility of a definitive trial to address the use of local gentamicin in open tibial fractures. If proven effective, local gentamicin would be a low-cost strategy to reduce complications and disability from open tibial fractures that could impact care in both high- and low-income countries. ClinicalTrials.gov, Registration # NCT03559400 ; Registered June 18, 2018.


The applications of machine learning algorithms in the modeling of estrogen-like chemicals.
Increasing concern is being shown by the scientific community, government regulators, and the public about endocrine-disrupting chemicals that, in the environment, are adversely affecting human and wildlife health through a variety of mechanisms, mainly estrogen receptor-mediated mechanisms of toxicity. Because of the large number of such chemicals in the environment, there is a great need for an effective means of rapidly assessing endocrine-disrupting activity in the toxicology assessment process. When faced with the challenging task of screening large libraries of molecules for biological activity, the benefits of computational predictive models based on quantitative structure-activity relationships to identify possible estrogens become immediately obvious. Recently, in order to improve the accuracy of prediction, some machine learning techniques were introduced to build more effective predictive models. In this review we will focus our attention on some recent advances in the use of these methods in modeling estrogen-like chemicals. The advantages and disadvantages of the machine learning algorithms used in solving this problem, the importance of the validation and performance assessment of the built models as well as their applicability domains will be discussed.


In vitro activity of retapamulin against Staphylococcus aureus isolates resistant to fusidic acid and mupirocin.
We determined the in vitro activity of retapamulin, a novel pleuromutilin antibiotic, against 664 Staphylococcus aureus isolates from the UK, including many resistant to fusidic acid and/or highly resistant to mupirocin. MICs were determined on Mueller-Hinton agar in accordance with the CLSI guidelines. Susceptibility was categorized using CLSI criteria, where available; otherwise the European Committee for Antimicrobial Susceptibility Testing (EUCAST)/BSAC criteria were used (for mupirocin and fusidic acid). Mutations in the rplC gene, which encodes ribosomal protein L3, were sought by PCR and DNA sequencing. The S. aureus included 488 (73%) methicillin-resistant isolates (oxacillin MICs >2 mg/L), 336 isolates (51%) resistant to fusidic acid (MICs >1 mg/L) and 254 (38%) with high-level mupirocin resistance (MICs >256 mg/L); 103 (16%) isolates were resistant both to fusidic acid and to high levels of mupirocin. Retapamulin inhibited 663 (99.9%) isolates at < or =0.25 mg/L. A single methicillin-resistant S. aureus isolate, also with high-level mupirocin resistance, required a retapamulin MIC of 2 mg/L, but its reduced susceptibility to retapamulin was not associated with any mutation in ribosomal protein L3. Retapamulin demonstrated excellent activity in vitro against S. aureus isolates, irrespective of their level of resistance to other antibacterials. These results support the EUCAST epidemiological cut-off value for retapamulin of < or =0.5 mg/L against S. aureus.


Novel NR2F1 variants likely disrupt DNA binding: molecular modeling in two cases, review of published cases, genotype-phenotype correlation, and phenotypic expansion of the Bosch-Boonstra-Schaaf optic atrophy syndrome.
Bosch-Boonstra-Schaaf optic atrophy syndrome (BBSOAS) is a recently described autosomal dominant disorder caused by mutations in the NR2F1 gene. There are presently 28 cases of BBSOAS described in the literature. Its common features include developmental delay, intellectual disability, hypotonia, optic nerve atrophy, attention deficit disorder, autism spectrum disorder, seizures, hearing defects, spasticity, and thinning of the corpus callosum. Here we report two unrelated probands with novel, de novo, missense variants in NR2F1 The first is a 14-yr-old male patient with hypotonia, intellectual disability, optic nerve hypoplasia, delayed bone age, short stature, and altered neurotransmitter levels on cerebrospinal fluid testing. The second is a 5-yr-old female with severe developmental delay, motor and speech delay, and repetitive motion behavior. Whole-exome sequencing identified a novel missense NR2F1 variant in each case, Cys86Phe in the DNA-binding domain in Case 1, and a Leu372Pro in the ligand-binding domain in Case 2. The presence of clinical findings compatible with BBSOAS along with structural analysis at atomic resolution using homology-based molecular modeling and molecular dynamic simulations, support the pathogenicity of these variants for BBSOAS. Short stature, abnormal CNS neurotransmitters, and macrocephaly have not been previously reported for this syndrome and may represent a phenotypic expansion of BBSOAS. A review of published cases along with new evidence from this report support genotype-phenotype correlations for this disorder.


Nonuniform blood flow in the canine left ventricle.
In order to investigate the relationship between coronary perfusion pressure and blood flow distribution in the left ventricle (LV), we measured myocardial blood flow in small regions using radioactive microspheres in six anesthetized, open-chest dogs. Mean coronary perfusion pressure (CPP) was controlled with a femoral artery to left main coronary artery shunt which included a pressurized, servo-controlled blood reservoir. In each dog, we measured flow in 192 regions of the LV free wall (mean weight per region = 206 +/- 38 mg) at different perfusion pressures. At CPP = 80 mm Hg, blood flow to individual regions varied fourfold (0.30 to 1.18 ml/min/g; relative dispersion (RD) = 21.8 +/- 2.3%). At CPP = 50 mm Hg, flow varied over sevenfold (0.08 to 0.60 ml/min/g; RD = 42.8 +/- 10%; P less than 0.01 vs 80 mm Hg). This relationship between flow variability and CPP was present within individual LV layers as well between layers and is much higher than the error associated with the microsphere technique. We conclude that blood flow to small regions of the LV is markedly nonuniform. This heterogeneity becomes more profound at lower CPP. These findings suggest that (1) global measurements of coronary flow must be interpreted with caution, and (2) even in hearts with normal coronary arteries some regions of the LV are more susceptible to ischemia than others. In addition, these findings may help explain the patchy nature of myocardial damage that occurs following periods of low coronary pressure or inadequate myocardial protection during cardiopulmonary bypass.


Effect of Low-Level Laser and Light-Emitting Diode on Inferior Alveolar Nerve Recovery After Sagittal Split Osteotomy of the Mandible: A Randomized Clinical Trial Study.
The major concern of sagittal split osteotomy (SSO) is the neurosensory disturbance. The authors investigated the effect of low-level laser therapy and light-emitting diode on the inferior alveolar nerve recovery after SSO. In this double-blinded randomized clinical trial, 20 subjects with inferior alveolar nerve injury after SSO of the mandible were studied. Neurosensory recovery was assessed by 6 tests: visual analog scale (VAS), brush stroke, 2-point discrimination, contact detect detection, pinprick nociception, and thermal discrimination, and each one was performed before and after the surgery on days 1, 3, 7, 14, 60, and 180. After 1 week, the VAS score in the laser group significantly improved in comparison with the control group. Visual analog scale score improvement was 25% (P = 0.015) at 2 weeks, 21% (P = 0.001) at 2 months, and 24% (P = 0.001) at 6 months. After 2 weeks, the brush stroke score improvement was significant in the laser group. The improvement values were 21.5% (P = 0.002) at 2 months and 15.1% (P = 0.004) at 6 months. Low-level laser therapy and light-emitting diode may improve VAS scores, 2-point discrimination, and brush stroke test results without any effect on the pinprick or contact detection test results.


[Oral aspects of porphyria].
Porphyria is a disease of specific enzymes in the biosynthesis of heme, caused by either genetic defects or environmental factors. This review of the literature presents the different types of porphyria and describes their possible causes, clinical signs, diagnosis and therapy. In addition, oral findings observed in patients with porphyria and the potential implications of the disease for dental treatment are discussed.


[Role of microviscosity of the lipid phase of microsomal membranes in regulation of enzymatic activity of glucose-6-phosphatase under microsomal modifications in vivo and in vitro].
The changes in microviscosity of the lipid phase of microsomal membranes under microsomal modification in vivo and in vitro were studied. It was shown that in intact microsome lipids there occur five thermo-induced structural transitions within the temperature range of 5--50 degrees. Delipidation of microsomes results in a shift in structural transitions temperature. Based on the literary and own data it was assumed that the breaks on the Arrhenius plots for glucose-6-phosphatase (EC 220.127.116.11) activity are due to phase-structural changes of microsomal lipids.


A Case of Radiation-Associated Vertebral Compression Fracture Mimicking Solitary Bone Metastasis of Lung Cancer.
Radiation therapy plays an important role in the treatment of lung cancer. Although adverse effects of radiation are well known, they are sometimes difficult to be diagnosed. We report a case of a radiation-associated vertebral compression fracture which mimicked bone metastasis of lung cancer. The patient was a 57-year-old man diagnosed with lung squamous cell carcinoma (cT1aN2M0, c-stage IIIA). He received concurrent chemoradiotherapy (CRT) in combination with 6 weeks of weekly carboplatin plus paclitaxel and thoracic radiation of 60 Gy/30 fractions, followed by bi-weekly durvalumab for 12 months. On the last day of the 12-month durvalumab regimen, he complained of backache. Magnetic resonance imaging showed compression fracture of the seventh thoracic vertebra with the spinal cord compressed, and fluorine-18 fluorodeoxyglucose positron emission tomography and computed tomography demonstrated weak focal uptake only at the seventh thoracic vertebra. Although the fracture had been suspected to be bone metastasis, surgical biopsy revealed no evidence of malignancy. Since the seventh thoracic vertebra was included in the irradiation area, the patient was diagnosed with a radiation-associated fracture. Dual-energy X-ray absorptiometry of the lumbar vertebrae (L2 - 4) after the surgery revealed osteopenia. In conclusion, we successfully diagnosed the radiation-associated vertebral fracture caused by radical CRT. The fracture mimicked bone metastasis in preoperative imaging tests. Thus, surgical biopsy was useful for diagnosis.


[Which tools should be used to identify learning objectives for the health service? The experience of the Emilia-Romagna Region (Italy)].
The main health education target of the Emilia-Romagna Region for the period 2008-2010 is to develop pathways of continuous learning for healthcare workers and managers in hospitals. The pathways have the aim of governing the innovation process, improving services, developing new skills, spreading good managerial and professional working practices and integrate knowledge and cooperation between professions at institutional and service levels. This paper synthetically illustrates the role of the different regional institutions (primary care trusts and NHS trusts) in establishing the objectives and the preferable procedures for teaching. The main difficulties concern the effective planning of homogeneous teaching, as well as the ways to translating the needs of the health of the patients into educational projects.


Stereospecific action of pyrethroid insecticides on the gamma-aminobutyric acid receptor-ionophore complex.
The potent alpha-cyano-3-phenoxybenzyl pyrethroids, including cypermethrin, deltamethrin, and fenvalerate, act stereospecifically to inhibit binding to rat brain synaptic membranes of sulfur-35-labeled t-butylbicyclophosphorothionate, a new radioligand for the picrotoxinin binding site. Scatchard analysis indicates that picrotoxinin inhibition of t-butylbicyclophosphorothionate binding is competitive whereas cypermethrin inhibition possibly involves a closely associated site in the gamma-aminobutyric acid receptor-ionophore complex. Studies with 37 pyrethroids reveal an absolute correlation, that is, no false positives or negatives, between mouse intracerebral toxicity and in vitro inhibition: all toxic cyano compounds but none of their nontoxic stereoisomers are inhibitors; cis isomers are more potent than trans isomers as both toxicants and inhibitors; and noncyano pyrethroids are much less potent or are inactive.


Pathophysiology, prevention, and treatment of commotio cordis.
Commotio cordis is increasing described and it is now clear that this phenomenon is an important cause of sudden cardiac death on the playing field. Victims are predominantly young, male, and struck in the left chest with a ball. An animal model has been developed and utilized to explore the important variables and mechanism of commotio cordis. Impact during a narrow window of repolarization causes ventricular fibrillation. Other important variables include location, velocity, shape, and hardness of the impact object. Biological characteristics such as gender, pliability of the chest wall, and genetic susceptibility also play a role in commotio cordis. The mechanism of ventricular fibrillation appears to be an increase in heterogeneity of repolarization caused by induced abnormalities of ion channels activated by abrupt increases in left ventricular pressure. In the setting of altered repolarization a trigger of ventricular depolarization (premature ventricular depolarization caused directly by the chest blow) initiates a spiral wave that quickly breaks down into ventricular fibrillation. Prevention of commotio cordis is possible. Improved recognition and resuscitation have led to an improvement in outcome.


Interaction of Leishmania gp63 with cellular receptors for fibronectin.
The most abundant protein on the surface of the promastigote form of the protozoan parasites Leishmania spp. is a 63-kDa molecule, designated gp63 or leishmanolysin. Because gp63 has been shown to possess fibronectin-like properties, we examined the interaction of gp63 with the cellular receptors for fibronectin. We measured the direct binding of Leishmania to human macrophages or to transfected mammalian cells expressing human fibronectin receptors. Leishmania expressing gp63 exhibited modest but reproducible adhesion to human macrophages and to transfected CHO cells expressing alpha4/beta1 fibronectin receptors. In both cases, this interaction depended on gp63 but occurred independently of the SRYD sequence of gp63, because parasites expressing gp63 with a mutated SRYD sequence bound to macrophages and alpha4/beta1 receptor-expressing cells as well as did wild-type parasites. The contribution of gp63 to parasite adhesion was more pronounced when the assays were performed in the presence of complement, suggesting that the receptors for complement and fibronectin may cooperate to mediate the efficient adhesion of parasites to macrophages. The interaction of gp63 with fibronectin receptors may also play an important role in parasite internalization by macrophages. Erythrocytes to which gp63 was cross-linked were efficiently phagocytized by macrophages, whereas control erythrocytes opsonized with complement alone bound to macrophages but remained peripherally attached to the outside of the cell. Similarly, parasites expressing wild-type gp63 were rapidly and efficiently phagocytized by resting macrophages, whereas parasites lacking gp63 were internalized more slowly. This rapid internalization of gp63-expressing parasites was dependent on the beta1 integrins, because pretreatment of macrophages with monoclonal antibodies to the beta1 integrins decreased the internalization of gp63-expressing parasites. These observations indicate that complement receptors are the primary mediators of parasite adhesion; however, maximal parasite adhesion and internalization may require the participation of the beta1 integrins, which recognize fibronectin-like molecules such as gp63 on the surface of the parasite.


Prevalence, Clinical Features, Neuroimaging, and Genetic Findings in Children With Ataxic Cerebral Palsy in Europe.
To report on prevalence, associated impairments, severity and neuroimaging findings in children with ataxic cerebral palsy (CP). In children coded as having ataxic CP in the Central database of JRC-SCPE (Joint Research Center-Surveillance of Cerebral Palsy in Europe) and born 1980-2010, birth characteristics, severity profiles including associated impairments, neuroimaging patterns and the presence of syndromes were analyzed. Definitions were according to validated SCPE guidelines. Prevalence over time was estimated using Poisson regression. In total, 679 children with ataxic CP were identified in 20 European CP registers. The proportion with ataxic CP was 3.8% and varied from 0% to 12.9%. Prevalence over time showed no significant trend. 70% of children with ataxic CP were able to walk, 40% had severe intellectual impairment and a high impairment index. Children with ataxic CP were mostly born at term (79%) and with normal birth weight (77%). Neuroimaging patterns revealed normal findings in 29%, brain maldevelopments in 28.5% and miscellaneous findings in 23.5% and brain injuries in 19%, according to the SCPE classification. Genetic syndromes were described in 9%. This register-based multicentre study on children with ataxic CP provides a large sample size for the analysis of prevalence, severity and origin of this rare CP subtype. Even with strict inclusion and classification criteria, there is variation between registers on how to deal with this subtype and diagnosis of ataxic CP remains a challenge. Ataxic cerebral palsy differs from other CP subtypes: children with ataxic CP have a disability profile that is more pronounced with respect to cognitive than gross motor dysfunction. They are mostly term born and the origin rarely suggests acquired injuries. In addition to neuroimaging, a comprehensive genetic work-up is particularly recommended for children with this CP type.


RNA-binding protein recognition based on multi-view deep feature and multi-label learning.
RNA-binding protein (RBP) is a class of proteins that bind to and accompany RNAs in regulating biological processes. An RBP may have multiple target RNAs, and its aberrant expression can cause multiple diseases. Methods have been designed to predict whether a specific RBP can bind to an RNA and the position of the binding site using binary classification model. However, most of the existing methods do not take into account the binding similarity and correlation between different RBPs. While methods employing multiple labels and Long Short Term Memory Network (LSTM) are proposed to consider binding similarity between different RBPs, the accuracy remains low due to insufficient feature learning and multi-label learning on RNA sequences. In response to this challenge, the concept of RNA-RBP Binding Network (RRBN) is proposed in this paper to provide theoretical support for multi-label learning to identify RBPs that can bind to RNAs. It is experimentally shown that the RRBN information can significantly improve the prediction of unknown RNA-RBP interactions. To further improve the prediction accuracy, we present the novel computational method iDeepMV which integrates multi-view deep learning technology under the multi-label learning framework. iDeepMV first extracts data from the views of amino acid sequence and dipeptide component based on the RNA sequences as the original view. Deep neural network models are then designed for the respective views to perform deep feature learning. The extracted deep features are fed into multi-label classifiers which are trained with the RNA-RBP interaction information for the three views. Finally, a voting mechanism is designed to make comprehensive decision on the results of the multi-label classifiers. Our experimental results show that the prediction performance of iDeepMV, which combines multi-view deep feature learning models with RNA-RBP interaction information, is significantly better than that of the state-of-the-art methods. iDeepMV is freely available at http://www.csbio.sjtu.edu.cn/bioinf/iDeepMV for academic use. The code is freely available at http://github.com/uchihayht/iDeepMV.


Innate immune activation by inhaled lipopolysaccharide, independent of oxidative stress, exacerbates silica-induced pulmonary fibrosis in mice.
Acute exacerbations of pulmonary fibrosis are characterized by rapid decrements in lung function. Environmental factors that may contribute to acute exacerbations remain poorly understood. We have previously demonstrated that exposure to inhaled lipopolysaccharide (LPS) induces expression of genes associated with fibrosis. To address whether exposure to LPS could exacerbate fibrosis, we exposed male C57BL/6 mice to crystalline silica, or vehicle, followed 28 days later by LPS or saline inhalation. We observed that mice receiving both silica and LPS had significantly more total inflammatory cells, more whole lung lavage MCP-1, MIP-2, KC and IL-1β, more evidence of oxidative stress and more total lung hydroxyproline than mice receiving either LPS alone, or silica alone. Blocking oxidative stress with N-acetylcysteine attenuated whole lung inflammation but had no effect on total lung hydroxyproline. These observations suggest that exposure to innate immune stimuli, such as LPS in the environment, may exacerbate stable pulmonary fibrosis via mechanisms that are independent of inflammation and oxidative stress.


Access to information supporting availability of medicines for patients suffering from rare diseases looking for possible treatments: the EuOrphan Service.
Currently in Europe, approximately 30 million people suffer from rare diseases, and a major problem is that many patients do not have access to quality healthcare for their disorders. Moreover, there is also a lack of quality information and a networking system aimed at supporting interaction among patients, clinicians, researchers, pharmaceutical industries, and governmental bodies. The purpose of this article is to inform physicians, public health care professionals, and other health care providers about EuOrphan service, the aim of which is to ensure easier access to quality information on rare diseases and their treatment. A set of web-based services is available at www.euorphan.com where information for target-users on treatments and products available worldwide for rare disease care as well as indications about healthcare centers are provided. Moreover, the service aims at providing consultancies for pharmaceutical companies to ultimately support the European legislation in bringing new drugs of a high ethical standard to the market and to exert a positive impact on the large population of patients suffering from rare diseases in Europe. The services provided by EuOrphan can facilitate concrete networking among patients, patient associations, doctors, and companies and also support the organization of clinical trials. In this perspective, EuOrphan could become a very valuable tool for globalizing the information about the availability of treatment (authorized or under development) of orphan patients.


Tyrosine phosphorylation of HSP90 within the P2X7 receptor complex negatively regulates P2X7 receptors.
The purinergic P2X7 receptor not only gates the opening of a cationic channel, but also couples to several downstream signaling events such as rapid membrane blebbing, microvesicle shedding, and interleukin-1beta release. Protein-protein interactions are likely to be involved in most of these signaling cascades; and recently, a P2X7 receptor-protein complex comprising at least 11 distinct proteins has been identified. We have studied one of these interacting proteins, HSP90, in human embryonic kidney cells expressing either human or rat P2X7 receptors as well as in rat peritoneal macrophages using biochemical (immunoprecipitation and Western blotting) and functional (membrane blebbing and currents) assays. We found that HSP90 was tyrosine-phosphorylated in association with the P2X7 receptor complex, but not in the cytosolic compartment. The HSP90 inhibitor geldanamycin decreased tyrosine phosphorylation of HSP90 and produced a 2-fold increase in the sensitivity of P2X7 receptors to agonist. Protein expression and tyrosine phosphorylation of a mutant P2X7 receptor in which a tyrosine in the C-terminal domain was substituted with phenylalanine (Y550F) were not changed, but tyrosine phosphorylation of HSP90 associated with this mutant P2X7 receptor complex was significantly greater than that associated with the wild-type complex. P2X7-Y550F receptors showed a 15-fold lower sensitivity to agonist, which was reversed by geldanamycin. We conclude that selective tyrosine phosphorylation of P2X7 receptor-associated HSP90 may act as a negative regulator of P2X7 receptor complex formation and function.


Confirmatory Factor Analysis of the Spirituality and Spiritual Care Rating Scale: A Cross-Sectional Study in Eight US Nursing Homes.
The aim of this study was to test a modified Spirituality and Spiritual Care Rating Scale (SSCRS) and report initial findings, based on a cross-sectional descriptive survey from eight nursing homes in the USA. This study examined the psychometric properties of a modified version of the SSCRS for assessing the perceptions of staff (i.e. nurses, rehabilitation staff, food and nutrition service staff, activities staff, social workers, and administrative staff) who work in nursing homes. The modified version of the SSCRS is measuring respondents' perceptions of spirituality, spiritual care, religiosity, and personalized care. Confirmatory factor analysis showed satisfactory goodness of fit for the original four-factor structure of the SSCRS. The modified version demonstrated good internal consistency and reliability (Cronbach's alpha ranging from 0.71 to 0.90). Initial findings showed statistically significant differences across all four sub-scales. Further research is needed to psychometrically test the modified tool.


Identification of DNA sequences and viral proteins of 6 human papillomavirus types in retinoblastoma tissue.
To detect DNA and proteins of human papillomavirus (HPV) in paraffin-embedded retinoblastoma (RB) tissue, to identify the viral types present and to describe a possible association between the presence of HPV and a particular form of RB. Fifty-one samples from ocular tissues of RB patients and of six controls enucleated for non-neoplastic reasons were obtained and analyzed by Polymerase Chain Reaction (PCR) with consensus primers to detect HPV. Viral type identification was performed by Restriction Fragment Length Polymorphisms (RFLP) analysis. To corroborate the presence of HPV, immunohistochemical analysis with a polyclonal anti-HPV antibody was performed in 10 RB cases and in all controls. Forty-two (82.3%) of the 51 samples were HPV-positive. HPV 6 was detected in 40 cases (95.2%), HPV 33 in 16 (38.1%), HPV 11 in 4 (9.5%) and HPV 31, 35 and 51 each in one case (2.3%). All controls were negative for HPV-DNA. The positive samples were PCR-tested for HPV 16 and 18 using specific primers, and were all negative. For immunohistochemical analysis, 7 out of 10 PCR-positive samples randomly chosen were positive; all six controls were negative. No differences in the HPV type distribution were found between the groups formed according to the tumor presentation or to the mode of inheritance.


Small-Cell Carcinoma of the Ovary, Hypercalcemic Type-Genetics, New Treatment Targets, and Current Management Guidelines.
Small-cell carcinoma of the ovary, hypercalcemic type (SCCOHT) is a rare and highly aggressive ovarian malignancy. In almost all cases, it is associated with somatic and often germline pathogenic variants in SMARCA4, which encodes for the SMARCA4 protein (BRG1), a subunit of the SWI/SNF chromatin remodeling complex. Approximately 20% of human cancers possess pathogenic variants in at least one SWI/SNF subunit. Because of their role in regulating many important cellular processes including transcriptional control, DNA repair, differentiation, cell division, and DNA replication, SWI/SNF complexes with mutant subunits are thought to contribute to cancer initiation and progression. Fewer than 500 cases of SCCOHT have been reported in the literature and approximately 60% are associated with hypercalcemia. SCCOHT primarily affects females under 40 years of age who usually present with symptoms related to a pelvic mass. SCCOHT is an aggressive cancer, with long-term survival rates of 30% in early-stage cases. Although various treatment approaches have been proposed, there is no consensus on surveillance and therapeutic strategy. An international group of multidisciplinary clinicians and researchers recently formed the International SCCOHT Consortium to evaluate current knowledge and propose consensus surveillance and therapeutic recommendations, with the aim of improving outcomes. Here, we present an overview of the genetics of this cancer, provide updates on new treatment targets, and propose management guidelines for this challenging cancer.


[CAR-T therapy for malignant lymphoma].
B-cell non-Hodgkin's lymphoma is a very heterogonous malignancy with diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) as the most common subtypes. Recent advances in chimeric antigen receptor T-cell (CAR-T) therapy are changing the current landscape for management of relapsed or refractory (R/R) DLBCL and R/R FL, which have a poor prognosis. Pivotal trials leading to the FDA approval of three CD19 CAR-T cells (Yescarta®, Kymriah® and Breyanzi®) showed complete response (CR) rates of 40-60%, with a significant subset of patients achieving long-term disease remission. Real-world studies have also confirmed this data. Notable toxicities include cytokine release syndrome and neurologic toxicities, which are usually treatable and reversible, as well as cytopenias and hypogammaglobulinemia. Salvage chemoimmunotherapy followed by high-dose chemotherapy and autologous stem cell rescue is the standard of care for chemo-sensitive and transplant-eligible R/R DLBCL. This review highlights the approved CAR-T constructs, including their efficacy, adverse effects, and real-world data.


Dietary intake and risk of rheumatoid arthritis-a cross section multicenter study.
Environmental factors play an important role in the development of rheumatoid arthritis (RA). Among these factors, smoking is generally considered to be an established risk factor for RA. Data regarding the impact of diet on risk of RA development is limited. This study assessed the impact of dietary patterns on RA susceptibility in Chinese populations. This was a large scale, case-control study composed of 968 patients with RA and 1037 matched healthy controls. Subjects were recruited from 18 teaching hospitals. Socio-demographic characteristics and dietary intakes 5 years prior to the onset of RA were reported by a self-administered questionnaire. Differences in quantity of consumption between cases and controls were analyzed by Student's t test. Multiple logistic regression analysis was applied to identify independent dietary risk factor(s) responsible for RA susceptibility. Compared to healthy individuals, RA patients had decreased consumption of mushrooms (P = 0.000), beans (P = 0.006), citrus (P = 0.000), poultry (P = 0.000), fish (P = 0.000), edible viscera (P = 0.018), and dairy products (P = 0.005). Multivariate analyses revealed that several dietary items may have protective effects on RA development, such as mushrooms (aOR = 0.669; 95%CI = 0.518-0.864, P = 0.002), citrus fruits (aOR = 0.990; 95%CI = 0.981-0.999, P = 0.04), and dairy products (aOR = 0.921; 95%CI 0.867-0.977, P = 0.006). Several dietary factors had independent effects on RA susceptibility. Dietary interventions may reduce the risk of RA.


Post-ECT Delirium.
A confusional state accompanied by restlessness is a frequent occurrence at the end of an induced seizure in electroconvulsive therapy (ECT). Delirium, which usually lasts <1 h, occurs in up to 12% of patients. It is most frequent during the initial treatments of a series. The occurrence of delirium in one treatment is not a harbinger of its reappearance in later treatments. Usually gentle reassurance is sufficient to help the patient, but some instances warrant the administration of diazepam, midazolam, a barbiturate, or droperidol.


Associations among behavioral inhibition and owner-rated attention, hyperactivity/impulsivity, and personality in the domestic dog (Canis familiaris).
In humans, behavioral disinhibition is associated with attention-deficit/hyperactivity disorder (ADHD). Limitations to rodent models of ADHD-like behaviors/symptoms may be augmented by complementary ones, such as the domestic dog. We examined associations between family dogs' (N = 29; of 14 breeds and 12 mongrels) performance on a self-developed touchscreen behavioral Go/No-Go paradigm and their owner-rated inattention and hyperactivity/impulsivity, accounting for relevant covariates. A greater proportion of commission errors was associated with greater hyperactivity/impulsivity. Regardless of accuracy, relative to dogs with no previous training, those with basic training had shorter response latencies. Also, regardless of accuracy, greater confidence and extraversion were associated with shorter latencies, and greater openness was associated with longer latencies. Shorter latency to commission errors was associated with greater inattention. Findings support the dog as a model of the association between behavioral disinhibition and ADHD-like behaviors/symptoms and are early evidence of convergent validity between the behavioral paradigm and the rating scale measure in dogs. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Using model-based screening to help discover unknown environmental contaminants.
Of the tens of thousands of chemicals in use, only a small fraction have been analyzed in environmental samples. To effectively identify environmental contaminants, methods to prioritize chemicals for analytical method development are required. We used a high-throughput model of chemical emissions, fate, and bioaccumulation to identify chemicals likely to have high concentrations in specific environmental media, and we prioritized these for target analysis. This model-based screening was applied to 215 organosilicon chemicals culled from industrial chemical production statistics. The model-based screening prioritized several recognized organosilicon contaminants and generated hypotheses leading to the selection of three chemicals that have not previously been identified as potential environmental contaminants for target analysis. Trace analytical methods were developed, and the chemicals were analyzed in air, sewage sludge, and sediment. All three substances were found to be environmental contaminants. Phenyl-tris(trimethylsiloxy)silane was present in all samples analyzed, with concentrations of ∼50 pg m(-3) in Stockholm air and ∼0.5 ng g(-1) dw in sediment from the Stockholm archipelago. Tris(trifluoropropyl)trimethyl-cyclotrisiloxane and tetrakis(trifluoropropyl)tetramethyl-cyclotetrasiloxane were found in sediments from Lake Mjøsa at ∼1 ng g(-1) dw. The discovery of three novel environmental contaminants shows that models can be useful for prioritizing chemicals for exploratory assessment.


Rapid Increase of the Serum PSA Level in Response to High-Intensity Focused Ultrasound Therapy may be a Potential Indicator of Biochemical Recurrence of Low- and Intermediate-Risk Prostate Cancer.
To determine the incidence and magnitude of the rapid increase in the serum PSA (riPSA) level after high-intensity focused ultrasound (HIFU) therapy for prostate cancer, and its correlation with clinical factors. A total of 176 patients with localized prostate cancer underwent HIFU therapy. Serum riPSA was determined on the basis of the same criteria as those for "PSA bounce", ie, an increase of ≥0.2 ng/ml with a spontaneous return to the prebounce level or lower. Patients were stratified according to neoadjuvant PSA level, T stage, risk group, age, Gleason score, pretreatment PSA level, post-treatment PSA nadir, and number of HIFU sessions. riPSA was seen in 53% of patients during a median follow-up period of 43 months. A PSA nadir was achieved within 3 months for 85.1% of the treatments. In all cases, onset of riPSA was seen two days after HIFU therapy, and the median magnitude was 23.69 ng/ml. A magnitude of >2 ng/ml was seen in 89.4% of cases. Univariate analysis revealed that patients with riPSA were associated with usage of hormonal therapy and the post-treatment PSA nadir level. Multivariate Cox regression analysis revealed that riPSA and the number of HIFU sessions were predictors of biochemical recurrence. A significant statistical association was found between the presence of riPSA and the risk of biochemical failure only in the low- and intermediate-risk group. Patients treated with HIFU who experience post-treatment riPSA may have an increased risk of biochemical recurrence, especially in non-high-risk patients.


Phase control of quasi-2D perovskites and improved light-emitting performance by excess organic cations and nanoparticle intercalation.
The optoelectronic properties of quasi-two-dimensional organic-inorganic hybrid perovskites can be tuned by controlling the formation of Ruddlesden-Popper type phases, which enables diverse device applications such as photovoltaics and light-emitting diodes (LEDs). Herein, the influence of excess organic cations on the phase formation of (PEA)2MAn-1PbnBr3n+1 is systematically investigated with various mixing ratios to discover the phase distribution beneficial for light-emitting diodes. It is found that PEA cations exceeding Pb ions in molar ratio are required to produce small-n phases in the films with a strong photoluminescence, while excess MA cations enable the formation of more large-n phases. Low electrical conductivity inherent to the properties of quasi-2D perovskites is further lowered by the introduction of excess organic cations. This is overcome by the intercalation of zinc oxide (ZnO) nanoparticles (NPs) into the blocking layers composed of PEA cations. Importantly, these metal oxide NPs also modulate the phase distribution, enabling the realization of bright green quasi-2D perovskites with a better stability and a maximum luminance of nearly 60 000 cd m-2, which is the highest brightness compared to the so far reported quasi-2D perovskite LEDs incorporating organic cations.


Visible light induced ocular delayed bioluminescence as a possible origin of negative afterimage.
The delayed luminescence of biological tissues is an ultraweak reemission of absorbed photons after exposure to external monochromatic or white light illumination. Recently, Wang, Bókkon, Dai and Antal (2011) [10] presented the first experimental proof of the existence of spontaneous ultraweak biophoton emission and visible light induced delayed ultraweak photon emission from in vitro freshly isolated rat's whole eye, lens, vitreous humor and retina. Here, we suggest that the photobiophysical source of negative afterimage can also occur within the eye by delayed bioluminescent photons. In other words, when we stare at a colored (or white) image for few seconds, external photons can induce excited electronic states within different parts of the eye that is followed by a delayed reemission of absorbed photons for several seconds. Finally, these reemitted photons can be absorbed by non-bleached photoreceptors that produce a negative afterimage. Although this suggests the photobiophysical source of negative afterimages is related retinal mechanisms, cortical neurons have also essential contribution in the interpretation and modulation of negative afterimages.


Residential air pollution does not modify the positive association between physical activity and lung function in current smokers in the ECRHS study.
Very few studies have examined whether a long-term beneficial effect of physical activity on lung function can be influenced by living in polluted urban areas. We assessed whether annual average residential concentrations of nitrogen dioxide (NO2) and particulate matter with aerodynamic diameters < 2.5 μm (PM2.5) and <10 μm (PM10) modify the effect of physical activity on lung function among never- (N = 2801) and current (N = 1719) smokers in the multi-center European Community Respiratory Health Survey. Associations between repeated assessments (at 27-57 and 39-67 years) of being physically active (physical activity: ≥2 times and ≥1 h per week) and forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC) were evaluated using adjusted mixed linear regression models. Models were conducted separately for never- and current smokers and stratified by residential long-term NO2, PM2.5 mass and PM10 mass concentrations (≤75th percentile (low/medium) versus >75th percentile (high)). Among current smokers, physical activity and lung function were positively associated regardless of air pollution levels. Among never-smokers, physical activity was associated with lung function in areas with low/medium NO2, PM2.5 mass and PM10 mass concentrations (e.g. mean difference in FVC between active and non-active subjects was 43.0 mL (13.6, 72.5), 49.5 mL (20.1, 78.8) and 49.7 mL (18.6, 80.7), respectively), but these associations were attenuated in high air pollution areas. Only the interaction term of physical activity and PM10 mass for FEV1 among never-smokers was significant (p-value = 0.03). Physical activity has beneficial effects on adult lung function in current smokers, irrespective of residential air pollution levels in Western Europe. Trends among never-smokers living in high air pollution areas are less clear.


Estimation of anisotropic permeability in trabecular bone based on microCT imaging and pore-scale fluid dynamics simulations.
In this paper, a comprehensive framework is proposed to estimate the anisotropic permeability matrix in trabecular bone specimens based on micro-computed tomography (microCT) imaging combined with pore-scale fluid dynamics simulations. Two essential steps in the proposed methodology are the selection of (i) a representative volume element (RVE) for calculation of trabecular bone permeability and (ii) a converged mesh for accurate calculation of pore fluid flow properties. Accurate estimates of trabecular bone porosities are obtained using a microCT image resolution of approximately 10 μm. We show that a trabecular bone RVE in the order of 2 × 2 × 2 mm3 is most suitable. Mesh convergence studies show that accurate fluid flow properties are obtained for a mesh size above 125,000 elements. Volume averaging of the pore-scale fluid flow properties allows calculation of the apparent permeability matrix of trabecular bone specimens. For the four specimens chosen, our numerical results show that the so obtained permeability coefficients are in excellent agreement with previously reported experimental data for both human and bovine trabecular bone samples. We also identified that bone samples taken from long bones generally exhibit a larger permeability in the longitudinal direction. The fact that all coefficients of the permeability matrix were different from zero indicates that bone samples are generally not harvested in the principal flow directions. The full permeability matrix was diagonalized by calculating the eigenvalues, while the eigenvectors showed how strongly the bone sample's orientations deviated from the principal flow directions. Porosity values of the four bone specimens range from 0.83 to 0.86, with a low standard deviation of ± 0.016, principal permeability values range from 0.22 to 1.45 ⋅ 10 -8 m2, with a high standard deviation of ± 0.33. Also, the anisotropic ratio ranged from 0.27 to 0.83, with high standard deviation. These results indicate that while the four specimens are quite similar in terms of average porosity, large variability exists with respect to permeability and specimen anisotropy. The utilized computational approach compares well with semi-analytical models based on homogenization theory. This methodology can be applied in bone tissue engineering applications for generating accurate pore morphologies of bone replacement materials and to consistently select similar bone specimens in bone bioreactor studies.


Sociophonetic Investigation of the Spanish Alveolar Trill /r/ in Two Canonical-Trill Varieties.
The "hyper-variation" present in rhotic sounds makes them particularly apt for sociophonetic research. This paper investigates the variable realization of the voiced alveolar-trill phoneme /r/ through an acoustic analysis of unscripted speech produced by 80 speakers of Spanish. Although the most common phonetic variant of /r/ contained two lingual constrictions, we find substantial inter-speaker variation in our data, ranging from zero to five lingual contacts. The results demonstrate that the variation in Spanish results from a systematic interaction of factors, deriving from well-documented processes of consonantal lenition (e.g., weakening in unstressed syllables) in addition to processes inherent to the trill's articulation (e.g., high-vowel antagonism). Importantly, speaker sex displayed the strongest effect among all the predictors, which leads us to consider the role of sociolinguistic factors, in addition to possible biomechanical differences, on /r/ production. We contextualize the findings within a literature that theorizes rhotic consonants as a single class of sounds despite remarkable patterns of cross-language and speaker-specific variation.


Assessment of trading partners for China's rare earth exports using a decision analytic approach.
Chinese rare earth export policies currently result in accelerating its depletion. Thus adopting an optimal export trade selection strategy is crucial to determining and ultimately identifying the ideal trading partners. This paper introduces a multi-attribute decision-making methodology which is then used to select the optimal trading partner. In the method, an evaluation criteria system is established to assess the seven top trading partners based on three dimensions: political relationships, economic benefits and industrial security. Specifically, a simple additive weighing model derived from an additive utility function is utilized to calculate, rank and select alternatives. Results show that Japan would be the optimal trading partner for Chinese rare earths. The criteria evaluation method of trading partners for China's rare earth exports provides the Chinese government with a tool to enhance rare earth industrial policies.


Associations between Hunger and Psychological Outcomes: A Large-Scale Ecological Momentary Assessment Study.
Studies assessing the association between hunger and psychological states have been conducted in laboratory settings, or limited to persons with eating disorders. In this study, 748 community-dwelling adults (56.4% women, 60.0 ± 9.3 years) completed the Ecological Momentary Assessment four times a day (08:00, 12:00, 16:00 and 20:00) for seven days. At each assessment, respondents indicated their current hunger level, food intake, and psychological states (sad, anxious, active, lively, distracted, anhedonic, angry, slow thinking and restless). Time-lagged associations assessing the effect of hunger on subsequent psychological states 4 h later and vice-versa were assessed. Hunger intensity increased subsequent active feeling (coefficient and 95% confidence interval: 0.029 (0.007; 0.051)) and lively feeling (0.019 (0.004; 0.034)) and decreased later slow thinking (-0.016 (-0.029; -0.003)). Previous eating increased later activity (0.116 (0.025; 0.208)). Feeling active (0.050 (0.036; 0.064)), lively (0.045 (0.023; 0.067)) and restless (0.040 (0.018; 0.063)) increased later hunger intensity, while distraction (-0.039 (-0.058; -0.019)) and slow thinking (-0.057 (-0.080; -0.034)) decreased it. No association was found between hunger, food intake and negative psychological states (sadness, anxiety and anger). Conclusions: Positive psychological states and hunger influence each other, while no association was found between hunger and negative psychological states.


[Fatal outcome of ischemic cerebral infarct].
During 1977 to 1985 2008 patients suffering from acute ischemic cerebral infarcts were admitted to the Department of Neurology, Giessen University. 213 (10.6%) died during their stay at hospital of 28 days in average. Time of survival, cause of death, localization and etiology of the infarcts were evaluated retrospectively on base of the medical reports, the angiographic, doppler-sonographic, computerized tomographic, and autopsy findings. The same number of patients died because of direct sequelae of stroke and secondary complications respectively. Those with supratentorial infarcts, who died in consequence of a vegetative dysregulation generally did not survive the first week after ictus, often had infarcts exceeding the supply area of one large cerebral artery and frequently had evidence of cardiac embolism. Pathogenetic factors for extension of the ischemic cerebral damage subsequently causing transtentorial herniation were spreading thrombosis, reinfarction, and fatal secondary hemorrhage. Patients dying in consequence of an infratentorial infarct often had a thrombosis of the basilar artery or a large cerebellar infarction. Secondary fatal complications mostly occurred after the first week after stroke. Pulmonary edema, pulmonary embolism and myocardial infarction predominated with different time pattern.


Bioanalysis of six antibiotics from volumetric microsamples: a new tool for precision dosing in critically ill children.
Background: Volumetric absorptive microsamples (VAMS) can support pharmacokinetic / pharmacodynamic studies. We present the bioanalytical method development for the simultaneous quantification of ampicillin, cefepime, ceftriaxone, meropenem, piperacillin, tazobactam, and vancomycin from VAMS. Methods & results: Optimal extraction, chromatographic, and mass spectrometry conditions were identified. Maximum extraction recoveries included 100 μl of water for rehydration and methanol for protein precipitation. Chromatographic separation used Phenomenex Kinetex™ Polar C18 column with a mobile phase comprising water/acetonitrile with formic acid and was fully validated. Hematocrit effects were only observed for vancomycin. Samples were stable for 90 days at -80°C except for meropenem, which was stable for 60 days. Conclusion: Multiple antibiotics can be assayed from a single VAMS sample to facilitate pharmacokinetic/pharmacodynamic studies.


Resonant spin wave excitation in magnetoplasmonic bilayers using short laser pulses.
In magnetically ordered solids a static magnetic field can be generated by virtue of the transverse magneto-optical Kerr effect (TMOKE). Moreover, the latter was shown to be dramatically enhanced due to the optical excitation of surface plasmons in nanostructures with relatively small optical losses. In this paper we suggest a new method for resonant optical excitation in a prototypical bilayer composed of a noble metal (Au) with grating and a ferromagnetic thin film of yttrium iron garnet (YIG) via a frequency comb. Based on magnetization dynamics simulations we show that for a frequency comb with certain parameters, chosen to be resonant with the spin-wave excitations of YIG, the TMOKE is drastically enhanced, hinting at possible technological applications in optical control of spintronics systems.


Salmonella promotes virulence by repressing cellulose production.
Cellulose is the most abundant organic polymer on Earth. In bacteria, cellulose confers protection against environmental insults and is a constituent of biofilms typically formed on abiotic surfaces. We report that, surprisingly, Salmonella enterica serovar Typhimurium makes cellulose when inside macrophages. We determine that preventing cellulose synthesis increases virulence, whereas stimulation of cellulose synthesis inside macrophages decreases virulence. An attenuated mutant lacking the mgtC gene exhibited increased cellulose levels due to increased expression of the cellulose synthase gene bcsA and of cyclic diguanylate, the allosteric activator of the BcsA protein. Inactivation of bcsA restored wild-type virulence to the Salmonella mgtC mutant, but not to other attenuated mutants displaying a wild-type phenotype regarding cellulose. Our findings indicate that a virulence determinant can promote pathogenicity by repressing a pathogen's antivirulence trait. Moreover, they suggest that controlling antivirulence traits increases long-term pathogen fitness by mediating a trade-off between acute virulence and transmission.


Splice Variants of pH-Sensitive Chloride Channel Identify a Key Determinant of Ivermectin Sensitivity in the Larvae of the Silkworm Bombyx mori.
The pH-sensitive chloride channels (pHCls) are broadly expressed in insects, but little is known about their physiologic role, diversity, and sensitivity to insecticides acting on relevant chloride channels. Here we have sequenced 50 transcripts of the pHCl-1 gene from the brain, third thoracic ganglion (T3G), and midgut of larvae of silkworm Bombyx mori It was found that >50 variants were expressed with distinct splicing in the T3G compared with the brain and midgut. Of the variants detected, variant 9, which was expressed most abundantly in the larvae, was reconstituted in Xenopus laevis oocytes to characterize its pH and ivermectin sensitivity. Variant 9 formed a functional pHCl with half-maximal activation at a pH of 7.87, and was activated by ivermectin irrespective of the extracellular pH. This was in contrast to variant 1, which was activated more profoundly at acidic rather than basic pH. To identify a key determinant for such differential ivermectin sensitivity, different amino acids in variants 1 and 9 were swapped, and the effects of the mutations on ivermectin sensitivity were investigated. The V275S mutation of variant 1 enhanced ivermectin sensitivity, whereas the S275V mutation of variant 9 caused a reduction in sensitivity. In homology models of the Bombyx pHCls, Val275 of variant 1 interacted more strongly with Ala273 than Ser275 of variant 9 at the channel gate. This interaction is likely to prevent ivermectin-induced opening of the channel, accounting, at least partially, for the differential macrolide action on the two variants.


Novel protein ADTRP regulates TFPI expression and function in human endothelial cells in normal conditions and in response to androgen.
Thrombosis and cardiovascular disease (CVD) represent major causes of morbidity and mortality. Low androgen correlates with higher incidence of CVD/thrombosis. Tissue Factor Pathway Inhibitor (TFPI) is the major inhibitor of tissue factor-factor VIIa (TF-FVIIa)-dependent FXa generation. Because endothelial cell (EC) dysfunction leading to vascular disease correlates with low EC-associated TFPI, we sought to identify mechanisms that regulate the natural expression of TFPI. Data mining of NCBI's GEO microarrays revealed strong coexpression between TFPI and the uncharacterized protein encoded by C6ORF105, which is predicted to be multispan, palmitoylated and androgen-responsive. We demonstrate that this protein regulates both the native and androgen-enhanced TFPI expression and activity in cultured ECs, and we named it androgen-dependent TFPI-regulating protein (ADTRP). We confirm ADTRP expression and colocalization with TFPI and caveolin-1 in ECs. ADTRP-shRNA reduces, while over-expression of ADTRP enhances, TFPI mRNA and activity and the colocalization of TF-FVIIa-FXa-TFPI with caveolin-1. Imaging and Triton X-114-extraction confirm TFPI and ADTRP association with lipid rafts/caveolae. Dihydrotestosterone up-regulates TFPI and ADTRP expression, and increases FXa inhibition by TFPI in an ADTRP- and caveolin-1-dependent manner. We conclude that the ADTRP-dependent up-regulation of TFPI expression and activity by androgen represents a novel mechanism of increasing the anticoagulant protection of the endothelium.


National food fortification: a dialogue with reference to Asia: policy in evolution.
Food fortification generally refers to the addition of micronutrients and other favourably bio-active food components to food-stuffs where there are recognised deficiencies in the target population. Each forticant has had or could have regulatory implications. It is understandable, although arguable, in the face of a limited food supply skewed, for the majority, in the direction of starchy staples of low essential nutrient density. Efforts, with plant breeding, to biofortify such foods are underway and likely to be safer, more sustainable and affordable than chemical additions. Unfortunately, with an increasingly refined and naturally tasteless food supply (salty, fatty, sugary and starchy), and where energy requirements are falling because of physical inactivity, micronutrient fortification is being used as a nutritional "fix-it' strategy. In Asia, there are several critical micro- nutrients. No one national fortification program can deal with all deficiencies is likely to be highly selective for the nutrients which have the greatest advocacy or are most recognisable. They also leave the other health promoting food properties like intactness, nutrient spectrum, and phytonutrient content un-addressed. A variety of food-stuffs, with different biological origins, is the preferred approach. Where an optimal food system is not in place, there may be justification for fortification if there is regular monitoring and surveillance of the food supply and health outcomes occurs; is a clear cost-risk-benefit advantage in such a strategy; are programs in place to improve the nutritional value of the basic food supply and is an "exit strategy' for the fortification program.


Magnetic resonance imaging compared with electrodiagnostic studies in patients with suspected carpal tunnel syndrome: predicting symptoms, function, and surgical benefit at 1 year.
The goal in this study of patients with clinical carpal tunnel syndrome (CTS) was to compare the usefulness of magnetic resonance (MR) imaging with that of electrodiagnostic studies (EDSs) for the following purposes: 1) prediction of 1-year outcomes and 2) identification of patients who are likely to benefit from surgical treatment. The authors prospectively enrolled 120 patients with clinically suspected CTS. The participants were tested using standardized EDSs, MR imaging, and a battery of questionnaires, including the Carpal Tunnel Syndrome Assessment Questionnaire, a well-validated 5-point score of symptoms and function. The EDSs and MR images were each interpreted independently. Patients were reevaluated after 1 year. The decision to treat patients conservatively or by carpal tunnel release was made by the individual surgeon, who had access to the initial EDS but not MR imaging results. Univariate and multivariate analyses were used to determine associations between 1-year outcomes and baseline diagnostic tests. The authors recontacted 105 of 120 participants at 12 months. Of these, 30 patients had had surgery and 75 had not. Patients who had undergone surgery showed greater improvement at 1 year than those who had not had surgery. The length of the abnormal T2-weighted nerve signal on MR imaging and median-ulnar sensory latency difference were the strongest predictors of surgical benefit. There was a clear patient preference for the MR imaging over EDSs. The findings obtained with MR imaging of the carpal tunnel predict surgical benefit independently of nerve conduction studies.


Evaluation of Mutual Drug-Drug Interaction within Geneva Cocktail for Cytochrome P450 Phenotyping using Innovative Dried Blood Sampling Method.
Cytochrome P450 (CYP) activity can be assessed using a 'cocktail' phenotyping approach. Recently, we have developed a cocktail (Geneva cocktail) which combines the use of low-dose probes with a low-invasiveness dried blood spots (DBS) sampling technique and a single analytical method for the phenotyping of six major CYP isoforms. We have previously demonstrated that modulation of CYP activity after pre-treatment with CYP inhibitors/inducer could be reliably predicted using Geneva cocktail. To further validate this cocktail, in this study, we have verified whether probe drugs contained in the latter cause mutual drug-drug interactions. In a randomized, four-way, Latin-square crossover study, 30 healthy volunteers received low-dose caffeine, flurbiprofen, omeprazole, dextromethorphan and midazolam (a previously validated combination with no mutual drug-drug interactions); fexofenadine alone; bupropion alone; or all seven drugs simultaneously (Geneva cocktail). Pharmacokinetic profiles of the probe drugs and their metabolites were determined in DBS samples using both conventional micropipette sampling and new microfluidic device allowing for self-sampling. The 90% confidence intervals for the geometric mean ratios of AUC metabolite/AUC probe for CYP probes administered alone or within Geneva cocktail fell within the 0.8-1.25 bioequivalence range indicating the absence of pharmacokinetic interaction. The same result was observed for the chosen phenotyping indices, that is metabolic ratios at 2 hr (CYP1A2, CYP3A) or 3 hr (CYP2B6, CYP2C9, CYP2C19, CYP2D6) post-cocktail administration. DBS sampling could successfully be performed using a new microfluidic device. In conclusion, Geneva cocktail combined with an innovative DBS sampling device can be used routinely as a test for simultaneous CYP phenotyping.


A matter of taste: Spatial and ontogenetic variations on the trophic ecology of the tiger shark at the Galapagos Marine Reserve.
Sharks are top predators across ocean food webs and have a major ecological role in marine ecosystems. Investigating the trophic ecology of this group of species is thus essential to understand ecosystem functioning and inform specific management actions aimed at shark conservation. The Galapagos Islands represent one of the last ocean wildernesses, where populations of sharks and other top marine predators come close to a pristine status. Here we provide the first study on the trophic ecology of the tiger shark (Galeocerdo cuvier) within the Galapagos Marine Reserve (GMR), using a combination of stable isotope analysis, satellite tracking, and passive acoustic telemetry to investigate ontogenetic and spatial variations at two regions. The mean estimated δ13C and δ15N at Isabela island (western region) were -13.9 ± 0.5‰ and 13.7 ± 0.7‰; and for Santa Cruz island (central region) were -13.8 ± 0.3‰ and 13.4 ± 0.7‰, respectively. Green sea turtles (Chelonia mydas) were the main prey item for large tiger sharks (>280 cm TL), while smaller sharks mainly fed on squid and pelagic fish. Tiger sharks exhibited a high degree of philopatry around green sea-turtle nesting areas, with the majority of sharks detected around green sea-turtle nesting areas for at least 10 months after their capture date, and some individuals were even present during the entire three-year study period. Although we did not report statistically significant differences between the two regions, isotopic and electronic tagging data suggest that tiger sharks in the Galapagos could be segregated into specific populations separated by geographical scales of <100 km. The high productivity of the archipelago, along with the protection from industrial fishing granted by the GMR, result in abundant and predictable sources of prey. This high food abundance, combined with the presence of suitable habitats throughout the tiger shark life cycle, might result in a reduction of migratory behaviours when compared to movement patterns of tiger sharks in other ocean basins. Additional studies using genetic tools could provide further evidence on the presence of separate management units, as it has been recently revealed for other shark species inhabiting the GMR.


Microinvasive carcinoma of the vulva. A report of two cases with regional lymph node metastasis.
Microinvasive squamous cell carcinoma of the vulva (invasion less than 5 mm) is a controversial concept. While initial observations did not identify regional lymph node metastases in patients with this lesion, 9.4% of all cases summarized from the literature have reported regional metastases. Presented here are two cases of poorly differentiated squamous cell carcinoma of the vulva (invasion of 2.0 and 3.0 mm) with lymph node metastases. Both patients' neoplasms consisted of a single focus of symmetrically expanding ("spray") carcinoma with infiltrating cords and isolated cells. While keratin formation was abundant, cytologic atypia was prominent. It is the authors' opinion that the degree of tumor differentiation, as judged by multiple parameters and pattern of invasion, is far more important than depth of invasion in predicting tumor behavior to determine therapy.


The obesity epidemic: can we turn the tide?
Obesity has reached epidemic proportions in the UK. It is important because of the associated co-morbidities, which include cardiovascular disease, type 2 diabetes, and osteoarthritis. The prevalence of obesity has increased because of a combination of excessive calorific intake (for example, from increased intake of energy dense foods) and insufficient energy expenditure (associated with a sedentary lifestyle). Weight loss of 5-10%, which can be achieved in primary care, is associated with significant health benefits. Obesity treatment in primary care includes lifestyle modification and drug treatment. The prevention and treatment of obesity cannot, however, be left solely to health professionals. Action is needed by government, the food industry, and society as a whole.


Bcl-2, Bcl-6, and the International Prognostic Index are prognostic indicators in patients with diffuse large B-cell lymphoma treated with rituximab-containing chemotherapy.
This study aimed to clarify the clinicopathological prognostic parameters of de novo diffuse large B-cell lymphoma (DLBCL) in the rituximab era. We examined the correlation of 22 clinicopathological parameters with progression-free survival (PFS), overall survival (OS), and primary refractory disease in 285 DLBCL patients treated with rituximab-containing chemotherapy. Complete response rate was 87%, overall response rate was 91%, 5-year PFS rate was 72%, and 5-year OS rate was 91%. By log-rank test, higher International Prognostic Index (IPI) (P < 0.0001), Bcl-2 positivity (P = 0.0013), Bcl-6 negativity (P = 0.0112), and no irradiation (P = 0.0371) were significantly correlated with shorter PFS; higher IPI (P = 0.0107), starry sky pattern (P = 0.0466), and no irradiation (P = 0.0264) correlated with shorter OS. In multivariate analyses, higher IPI (P = 0.0006), Bcl-2 positivity (P = 0.0015), and Bcl-6 negativity (P = 0.04) were significantly correlated with shorter PFS; higher IPI (P = 0.0045) correlated with shorter OS. Bcl-2 (P = 0.0029), Bcl-6 (P = 0.002), and IPI (P < 0.0001) were significantly correlated with primary refractory disease. In conclusion, Bcl-2 positivity, Bcl-6 negativity, and higher IPI were indicators of shorter PFS and OS plus primary refractory disease in patients with DLBCL in the rituximab era.


Lung function impairment and risk of incident heart failure: the NHLBI Pooled Cohorts Study.
The aim is to evaluate associations of lung function impairment with risk of incident heart failure (HF). Data were pooled across eight US population-based cohorts that enrolled participants from 1987 to 2004. Participants with self-reported baseline cardiovascular disease were excluded. Spirometry was used to define obstructive [forced expiratory volume in 1 s/forced vital capacity (FEV1/FVC) <0.70] or restrictive (FEV1/FVC ≥0.70, FVC <80%) lung physiology. The incident HF was defined as hospitalization or death caused by HF. In a sub-set, HF events were sub-classified as HF with reduced ejection fraction (HFrEF; EF <50%) or preserved EF (HFpEF; EF ≥50%). The Fine-Gray proportional sub-distribution hazards models were adjusted for sociodemographic factors, smoking, and cardiovascular risk factors. In models of incident HF sub-types, HFrEF, HFpEF, and non-HF mortality were treated as competing risks. Among 31 677 adults, there were 3344 incident HF events over a median follow-up of 21.0 years. Of 2066 classifiable HF events, 1030 were classified as HFrEF and 1036 as HFpEF. Obstructive [adjusted hazard ratio (HR) 1.17, 95% confidence interval (CI) 1.07-1.27] and restrictive physiology (adjusted HR 1.43, 95% CI 1.27-1.62) were associated with incident HF. Obstructive and restrictive ventilatory defects were associated with HFpEF but not HFrEF. The magnitude of the association between restrictive physiology and HFpEF was similar to associations with hypertension, diabetes, and smoking. Lung function impairment was associated with increased risk of incident HF, and particularly incident HFpEF, independent of and to a similar extent as major known cardiovascular risk factors.


Teledermatology in India: practical implications.
Teledermatology is considered to be one of the solutions to the problem of inadequate number of dermatologists in remote parts of India. We present a brief account of the technological components involved in teledermatology in India, and an evaluation of the advantages and limitations of the use of teledermatology as a clinical tool.


Embryology of the sternum.
Development of the sternum during the growth of the embryo is described. Anterior body wall defects in the thoracic region may be severe, leading to ectopia cordis, or mild, as in skin-covered sternal clefts. The embryologic basis for other sternal abnormalities, such as pectus excavatum and pectus carniatum, is not clear; however, abnormalities of rib morphogenesis and growth are the most likely causes.


[Value of the de Kleyn symptom in diagnosing vertebro-basilar insufficiency].
A total of 540 cases with cerebral vascular disorders examined otoneurologically were analyzed. The observed patients were treated in the Research Institute of Neurology. In 62 patients (11,5%) the De Klein symptom was detected. A comparison of the otoneurological rheoencephalographic and neurological symptomatology gives grounds to evaluate the De Klein symptom, encountered in different degrees of circulatory insufficiency in the vertebro-basillar system, as one of the early signs of vascular decompensation in initial forms of cerebro-vascular insufficiency, marking its diagnostic significance.


Sub-threshold mental health syndromes: finding an alternative to the medication of unhappiness.
Sub-threshold anxiety and depression are common presentations in primary care. They carry a significant disability burden along with the risk of developing a frank disorder. Intervention options are limited, although there is some evidence that ultra brief interventions may be effective with this patient group. We argue that there is a need for a systematic but ultra brief, minimal contact intervention, that can be delivered by GPs or practice nurses. Such an intervention would be a form of facilitated self-management, a step up from self-help, from which people could be referred on to more intensive treatment or medication if required.


Atm-deficient mice exhibit increased sensitivity to dextran sulfate sodium-induced colitis characterized by elevated DNA damage and persistent immune activation.
The role of ataxia telangiectasia mutated (ATM), a DNA double-strand break recognition and response protein, in inflammation and inflammatory diseases is unclear. We have previously shown that high levels of systemic DNA damage are induced by intestinal inflammation in wild-type mice. To determine the effect of Atm deficiency in inflammation, we induced experimental colitis in Atm(-/-), Atm(+/-), and wild-type mice via dextran sulfate sodium (DSS) administration. Atm(-/-) mice had higher disease activity indices and rates of mortality compared with heterozygous and wild-type mice. Systemic DNA damage and immune response were characterized in peripheral blood throughout and after three cycles of treatment. Atm(-/-) mice showed increased sensitivity to levels of DNA strand breaks in peripheral leukocytes, as well as micronucleus formation in erythroblasts, compared with heterozygous and wild-type mice, especially during remission periods and after the end of treatment. Markers of reactive oxygen and nitrogen species-mediated damage, including 8-oxoguanine and nitrotyrosine, were present both in the distal colon and in peripheral leukocytes, with Atm(-/-) mice manifesting more 8-oxoguanine formation than wild-type mice. Atm(-/-) mice showed greater upregulation of inflammatory cytokines and significantly higher percentages of activated CD69+ and CD44+ T cells in the peripheral blood throughout treatment. ATM, therefore, may be a critical immunoregulatory factor dampening the deleterious effects of chronic DSS-induced inflammation, necessary for systemic genomic stability and homeostasis of the gut epithelial barrier.


Prostate cancer laparoscopic port site metastasis: A case report.
Laparoscopic port site metastases are a rare but clinically important cause of biochemically recurrent prostate adenocarcinoma. C-11 choline, among other prostate-specific positron emission tomography (PET) radiotracers, has improved radiologist confidence in these otherwise difficult-to-detect sites of recurrence. A 62-year-old male presented with biochemically recurrent prostate adenocarcinoma after undergoing robotic-assisted radical prostatectomy 5 years earlier. C-11 choline positron emission tomography/computed tomography (PET/CT) demonstrated a choline-avid soft tissue nodule associated with a laparoscopic port site in the right rectus abdominis muscle, with correlative findings on prior magnetic resonance imaging, and biopsy confirming a prostate adenocarcinoma metastasis. The patient was initiated on chemohormonal therapy. His prostate-specific antigen (PSA) became undetectable following chemohormonal therapy. A follow-up C-11 choline PET/CT demonstrated complete resolution of prior abnormal radiotracer activity in the right rectus abdominis muscle. Port site metastases in prostate adenocarcinoma are rare; however, those who treat prostate cancer patients should be aware of this phenomenon as the number of minimally invasive oncologic procedures increase. C-11 choline PET, among other prostate-specific PET probes, has become an important tool in evaluating patients with biochemically recurrent prostate adenocarcinoma, identifying site-specific metastases in a majority of patients.


Wounds of the abdomen. Part 2: In peace.
Until the mid 1960s in the United States, penetrating abdominal wounds in civilians were badly handled and often fatal. Little attention was paid to the methods used and results reported by military surgeons in previous wars such as those in Korea and Vietnam. In 1966 the National Academy of Sciences, National Research Council Committee on Shock pointed out that there was an increasing incidence of severe civilian injuries associated with unacceptably high morbidity and mortality and that this was because of lack of organized civilian community care. As a result the Trauma/Emergency Medical Service (T/EMS) was formed. The initiators based the organization on the remarkable success of military concepts and techniques. The improvement in the evacuation and treatment of penetrating abdominal wounds in peace from the inception of the T/EMS to the present is described in this paper. The methods used and results obtained through the T/EMS have made an important contribution to both civilian and military surgery--a unique event in peacetime.


Diagnostic Utility of Exome Sequencing for Kidney Disease.
Exome sequencing is emerging as a first-line diagnostic method in some clinical disciplines, but its usefulness has yet to be examined for most constitutional disorders in adults, including chronic kidney disease, which affects more than 1 in 10 persons globally. We conducted exome sequencing and diagnostic analysis in two cohorts totaling 3315 patients with chronic kidney disease. We assessed the diagnostic yield and, among the patients for whom detailed clinical data were available, the clinical implications of diagnostic and other medically relevant findings. In all, 3037 patients (91.6%) were over 21 years of age, and 1179 (35.6%) were of self-identified non-European ancestry. We detected diagnostic variants in 307 of the 3315 patients (9.3%), encompassing 66 different monogenic disorders. Of the disorders detected, 39 (59%) were found in only a single patient. Diagnostic variants were detected across all clinically defined categories, including congenital or cystic renal disease (127 of 531 patients [23.9%]) and nephropathy of unknown origin (48 of 281 patients [17.1%]). Of the 2187 patients assessed, 34 (1.6%) had genetic findings for medically actionable disorders that, although unrelated to their nephropathy, would also lead to subspecialty referral and inform renal management. Exome sequencing in a combined cohort of more than 3000 patients with chronic kidney disease yielded a genetic diagnosis in just under 10% of cases. (Funded by the National Institutes of Health and others.).


Role of serotonergic input in the regulation of the beta-adrenergic receptor-coupled adenylate cyclase system.
The action of desipramine on the norepinephrine-sensitive adenylate cyclase system and the density of beta-adrenergic receptors in rat cortex was studied after selective lesioning of serotonergic neurons with 5,7-dihydroxytryptamine. In animals with lesions desipramine failed to reduce the density of beta-adrenoceptors but decreased the response of adenosine 3',5'-monophosphate to isoproterenol and norepinephrine to the same degree as in animals without lesions. The results demonstrate a functional linkage between serotonergic and noradrenergic systems in the rat cortex, with beta-adrenergic receptors and neurohormonal sensitivity of the adenosine 3',5'-monophosphate-generating system being under separate regulatory control.


Association of pain and CNS structural changes after spinal cord injury.
Traumatic spinal cord injury (SCI) has been shown to trigger structural atrophic changes within the spinal cord and brain. However, the relationship between structural changes and magnitude of neuropathic pain (NP) remains incompletely understood. Voxel-wise analysis of anatomical magnetic resonance imaging data provided information on cross-sectional cervical cord area and volumetric brain changes in 30 individuals with chronic traumatic SCI and 31 healthy controls. Participants were clinically assessed including neurological examination and pain questionnaire. Compared to controls, individuals with SCI exhibited decreased cord area, reduced grey matter (GM) volumes in anterior cingulate cortex (ACC), left insula, left secondary somatosensory cortex, bilateral thalamus, and decreased white matter volumes in pyramids and left internal capsule. The presence of NP was related with smaller cord area, increased GM in left ACC and right M1, and decreased GM in right primary somatosensory cortex and thalamus. Greater GM volume in M1 was associated with amount of NP. Below-level NP-associated structural changes in the spinal cord and brain can be discerned from trauma-induced consequences of SCI. The directionality of these relationships reveals specific changes across the neuroaxis (i.e., atrophic changes versus increases in volume) and may provide substrates of underlying neural mechanisms in the development of NP.


Aortic Wall Elastic Properties in Case of Bicuspid Aortic Valve.
Bicuspid aortic valve (BAV) is associated with a significant risk of development of aneurysm and dissection of the ascending thoracic aorta. Development of what is called BAV associated aortopathy is particularly heterogeneous with an uncertain prognosis and with no prognostic biomarkers except for the aortic diameter. This situation leads to an important variability of the therapeutic strategy of this aortopathy. By reviewing the literature on aortic stiffness in the case of BAV, we aimed at evaluating its potential prognostic role in the development of aortic dilatation. Studies evaluating aortic stiffness, with ultrasound or magnetic resonance imaging, converge toward the description of an increased segmental aortic stiffness in BAV patients regardless of age, diameter or aortic level, from the root to the arch. Even though there is a lack of longitudinal studies evaluating the progression of aortic dilatation, new data have recently shown the potential prognostic role of the maximal rate of systolic distension of the aortic wall with magnetic resonance imaging. Although the use of aortic distensibility calculation is a simple evaluation of stiffness that could be easily transposed in daily practice, its interpretation remains uncertain. New arterial stiffening indicators seem more promising but need a stronger validation.


Haematopoietic leptin receptor deficiency does not affect macrophage accumulation in adipose tissue or systemic insulin sensitivity.
The adipokine leptin is primarily produced by white adipose tissue (AT) and is a potent monocyte/macrophage chemoattractant in vitro. The long form of the leptin receptor (LepR) is required for monocyte/macrophage chemotaxis towards leptin. In this study, we examined the effects of haematopoietic LepR as well as LepR with C-C chemokine receptor 2 (CCR2) deficiency (double knockout (DKO)) on macrophage recruitment to AT after two different periods of high fat diet (HFD) feeding. Briefly, 8-week-old C57BL/6 mice were transplanted with bone marrow (BM) from Lepr(+/+), Lepr(-/-) or DKO donors (groups named BM-Lepr(+/+), BM-Lepr(-/-) and BM-DKO respectively), and were placed on an HFD for 6 or 12 weeks. At the end of the study, macrophage infiltration and the inflammatory state of AT were evaluated by real-time RT-PCR, histology and flow cytometry. In addition, glucose and insulin tolerance were assessed at both time points. Our results showed no differences in macrophage accumulation or AT inflammatory state between the BM-Lepr(+/+) and BM-Lepr(-/-) mice after 6 or 12 weeks of HFD feeding; any effects observed in the BM-DKO were attributed to the haematopoietic deficiency of CCR2. In addition, no changes in glucose or insulin tolerance were observed between groups after either period of HFD feeding. Our findings suggest that although leptin is a potent chemoattractant in vitro, haematopoietic LepR deficiency does not affect macrophage accumulation in AT in early to moderate stages of diet-induced obesity.


Humeral Retroversion (Complexity of Assigning Reference Axes in 3D and Its Influence on Measurement): A Technical Note.
Humeral retroversion (RV) is important to the study of shoulder function and reconstruction. This study tests the hypothesis that clinically obtained computer tomography (CT) measurements for humeral RV (off-axis measurements) differ from those obtained after reformatting the image slice orientation so that the humeral shaft is perpendicular to the gantry (coaxial measurements) and explores deviations from true RV. A custom-built application created in Mathematica was used to explore the effect of altering the humeral orientation on slice angle acquisition by 3D imaging technologies, on the perceived angle of RV from the 2D-projection of the reference axes. The application allows for control of humeral axis orientation relative to image slice (3D) or plain of projection (2D) and humeral rotation. The effect of rotating a virtual model of one humerus around its own axis and in discrete anatomical directions on the measured RV angle was assessed. The coaxial measurement of humeral RV (31.2°) differed from off-axis measurement, with a maximum difference in measured RV of 50° in 45° of extension. The typical position of the humerus in a CT scan resulted in a difference in RV measurement up to 22°. Explorations of deviation led to the following outcomes, as divided by anatomic direction. Extension and abduction led to an underestimation, and flexion and adduction led to an overestimation of the RV-angle. Measurements must be done consistently about the position and orientation of the humerus. Deviation in the humeral alignment of as little as 10° can distort the measurement of version up to 15°. van de Bunt F, Pearl ML, van Noort A. Humeral Retroversion (Complexity of Assigning Reference Axes in 3D and Its Influence on Measurement): A Technical Note. Strategies Trauma Limb Reconstr 2020;15(2):69-73.


Bemisia tabaci: a statement of species status.
Bemisia tabaci has long been considered a complex species. It rose to global prominence in the 1980s owing to the global invasion by the commonly named B biotype. Since then, the concomitant eruption of a group of plant viruses known as begomoviruses has created considerable management problems in many countries. However, an enduring set of questions remains: Is B. tabaci a complex species or a species complex, what are Bemisia biotypes, and how did all the genetic variability arise? This review considers these issues and concludes that there is now sufficient evidence to state that B. tabaci is not made up of biotypes and that the use of biotype in this context is erroneous and misleading. Instead, B. tabaci is a complex of 11 well-defined high-level groups containing at least 24 morphologically indistinguishable species.


The effect of orally administered probiotic Lactobacillus reuteri-containing tablets in peri-implant mucositis: a double-blind randomized controlled trial.
Probiotics create a biofilm and protect the oral tissues against the action of periodontal pathogenic bacteria. The aim of this study was to evaluate the effects of the oral probiotic Lactobacillus reuteri Prodentis upon the peri-implant health of edentulous patients with dental implants and peri-implant mucositis, establishing comparisons vs implants without peri-implant disease. A double-blind, placebo-controlled, prospective cross-over study was made. The patients were all edentulous and were divided into two groups, (A) no peri-implant disease, and (B) peri-implant mucositis affecting one or more implants. Patients with peri-implantitis were excluded. The dosage was one tablet every 24 h over 30 d. All patients in both groups initially received the oral probiotic Lactobacillus reuteri Prodentis, followed by placebo. Patients started with probiotic treatment during 30 d, followed by a 6 mo washout period and the administration of placebo for the same period. The following parameters were studied: crevicular fluid volume, modified plaque index, probing depth, modified gingival index, and concentrations of interleukin 1β, interleukin 6 and interleukin 8. A total of 77 implants were evaluated in 34 patients. Group A involved 22 patients with 54 implants without peri-implant alterations, and group B, 12 patients with mucositis affecting one or more implants (23 implants). After treatment with the probiotic, both the patients with mucositis and the patients without peri-implant disease showed improvements in the clinical parameters, with reductions in cytokine levels. In contrast, no such changes were observed with placebo. After treatment with the probiotic Lactobacillus reuteri in patients with implants presenting mucositis, the clinical parameters improved, and the cytokine levels decreased - in contraposition to the observations in the placebo group. Probiotic administration may be regarded as a good alternative for both the treatment of peri-implant mucositis and its prevention, as it also improved clinical parameters in the healthy individuals. Further studies involving larger patient series are needed regarding the effects of probiotics upon peri-implant health.


Human papillomavirus and posttransplantation cutaneous squamous cell carcinoma: A multicenter, prospective cohort study.
Organ transplant recipients (OTRs) have a 100-fold increased risk of cutaneous squamous cell carcinoma (cSCC). We prospectively evaluated the association between β genus human papillomaviruses (βPV) and keratinocyte carcinoma in OTRs. Two OTR cohorts without cSCC were assembled: cohort 1 was transplanted in 2003-2006 (n = 274) and cohort 2 was transplanted in 1986-2002 (n = 352). Participants were followed until death or cessation of follow-up in 2016. βPV infection was assessed in eyebrow hair by using polymerase chain reaction-based methods. βPV IgG seroresponses were determined with multiplex serology. A competing risk model with delayed entry was used to estimate cumulative incidence of histologically proven cSCC and the effect of βPV by using a multivariable Cox regression model. Results are reported as adjusted hazard ratios (HRs). OTRs with 5 or more different βPV types in eyebrow hair had 1.7 times the risk of cSCC vs OTRs with 0 to 4 different types (HR 1.7, 95% confidence interval 1.1-2.6). A similar risk was seen with high βPV loads (HR 1.8, 95% confidence interval 1.2-2.8). No significant associations were seen between serum antibodies and cSCC or between βPV and basal cell carcinoma. The diversity and load of βPV types in eyebrow hair are associated with cSCC risk in OTRs, providing evidence that βPV is associated with cSCC carcinogenesis and may present a target for future preventive strategies.


Long-term followup of the ventrally placed buccal mucosa onlay graft in bulbar urethral reconstruction.
We investigate whether the short-term success rate (greater than 90%) of buccal mucosa free grafts in the bulbar urethra is sustained in the long term. In 60 patients a ventrally placed buccal mucosa graft was used for repair of bulbar urethral strictures. Of these patients 49 had undergone previous attempt at repair (urethroplasty in 4, internal urethrotomy in 45). Mean graft length was 4.8 cm. In 9 patients a distal penile fasciocutaneous flap was also used for repair of concomitant penile urethral stricture. In 8 of the 9 patients the buccal mucosa graft was combined with end-to-end urethroplasty and 2 buccal mucosa grafts were used in tandem in 1. Followup was at least 1 year in all cases (mean 47 months, range 12 to 107). Failure was defined as an obstructive voiding pattern with radiographic or cystoscopic evidence of recurrent stricture. Bulbar stricture repair was successful in 54 patients (90%) and 4 of the remaining 6 responded to 1 internal urethrotomy for a long-term success rate of 97%. Preoperative clinical characteristics were not significantly different between those who experienced success or failure. Long-term outcome analysis of ventrally placed buccal mucosa onlay grafts for bulbar urethral strictures demonstrates a durable success rate of 90%. This rate can be improved (97%) with the judicious use of internal urethrotomy.


A New Biosensor for Stilbenes and a Cannabinoid Enabled by Genome Mining of a Transcriptional Regulator.
In vivo biosensors are powerful tools for metabolic engineering and synthetic biology applications. However, the development of biosensors is hindered by the limited number of characterized transcriptional regulators. The versatile sensing abilities of microbes and genome sequences available hold great potential for developing novel biosensors via genome mining for new transcriptional regulators. Here we report the development and engineering of a new stilbene-responsive biosensor discovered by mining the Novosphingobium aromaticivorans DSM 12444 genome. The biosensor can distinguish resveratrol from its precursors, p-coumaric acid and trans-cinnamic acid. Remarkably, it can detect other biologically active stilbenes with resorcinol groups, and cannabidiolic acid with a β-resorcylic acid functional group. When coupled to resveratrol biosynthesis enzymes, the biosensor can sense altered resveratrol production in cells, demonstrating a 667-fold enrichment in one round of fluorescence-activated cell sorting. Our biosensor will be potentially applicable to metabolic engineering of microbial cell factories for production of stilbenes and cannabinoids.