Structural characterization of OxyD, a cytochrome P450 involved in beta-hydroxytyrosine formation in vancomycin biosynthesis.
The cytochrome P450 OxyD from the balhimycin glycopeptide antibiotic biosynthetic operon of Amycolatopsis mediterranei is involved in the biosynthesis of the modified amino acid beta-R-hydroxytyrosine, an essential precursor for biosynthesis of the vancomycin-type aglycone. OxyD binds the substrate tyrosine not free in solution, but rather covalently linked to the carrier protein (CP) domain of the non-ribosomal peptide synthase BpsD, exhibiting micromolar binding affinity to a tyrosine-loaded carrier protein construct. The crystal structure of OxyD was determined to 2.1-A resolution, revealing a potential binding site for the carrier protein-bound substrate in a different orientation to that seen with the acyl carrier protein-bound P450(BioI) (Cryle, M. J., and Schlichting, I. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 15696-15701). A series of residues were identified across known aminoacyl-CP-oxidizing P450s that are highly conserved and cluster in the active site or potential CP binding site of OxyD. These residues appear to be characteristic for aminoacyl-CP-oxidizing P450s, allowing sequence based identification of P450 function for this subgroup of P450s that play vital roles in the biosyntheses of many important natural products in addition to the vancomycin-type antibiotics. The ability to analyze such P450 function based upon sequence data alone should prove an important tool in the analysis and identification of new medicinally relevant biomolecules.


Individualised radial extracorporeal shock wave therapy (rESWT) for symptomatic calcific shoulder tendinopathy: a retrospective clinical study.
A retrospective single centre cohort analysis was performed to evaluate an individualised radial extracorporeal shock wave therapy (rESWT) protocol for treatment of symptomatic calcific shoulder tendinopathy. 67 patients (79 Shoulders) were identified with 76 shoulders included for analysis. rESWT treatment protocol was adapted according to individual response to treatment. Variables included number of sessions, shockwave impulses, pressure and frequency. Success rate was estimated as the percentage of patients having ≥60% visual analogue score (VAS) pain decrease at follow-up. Recurrence at 1 year was recorded. Using this individualised symptom guided protocol, patients underwent a mean of 7 ± 1.5 rESWT sessions, with mean pressure of 1.7 ± 0.2 bar, mean frequency of 5 ± 0.3 Hz and 2175 ± 266 impulses. The mean pre-treatment VAS score of 6.7 ± 1.1 was significantly decreased to 3.2 ± 0.8 immediately post-treatment, 2.6 ± 0.9 at 1 month, 1.7 ± 1.0 at 3 months and 0.8 ± 1.0 at 1 year follow up (α = 0.05). One-year success rate was estimated at 92% and 1-year recurrence rate was 7%. We conclude that in this retrospective study an individualised rESWT protocol resulted in a high success rate with low number of recurrences. Randomised controlled trials to support these findings are recommended.


Oligomerisation of pneumolysin on cholesterol crystals: similarities to the behaviour of polyene antibiotics.
Pneumolysin is a cytolytic toxin of Streptococcus pneumoniae, a causative agent of pneumonia and meningitis. The prepore and pore states of pneumolysin have recently been investigated by cryo-electron microscopy and atomic force microscopy, confirming the existence of arc-shaped as well as ring-form oligomers. Here we provide further insights into the pneumolysin oligomer by studying the interaction of pneumolysin with cholesterol crystals, comparing the results to those obtained for polyene antibiotics, which also bind cholesterol.


Phenotypic and functional comparison between flask adherent and magnetic activated cell sorted monocytes derived dendritic cells.
Generation of an effective dendritic cell (DC) based cancer vaccine depends on appropriate differentiation of monocytes in vitro. To compare the effects of monocyte separation methods, flask adherence (Flask-DC) and magnetic activated cell sorting (MACS-DC), on phenotypic and functional characteristics of resultant DCs. DCs from healthy volunteers were generated from plastic adherence and MACS isolated monocytes in the presence of GM-CSF and IL-4 as well as TNF-α and monocyte conditioned medium (MCM) in 7 day cultures. Mature DCs were then subjected to phenotypic analysis using anti-CD14, anti-CD83 and HLA-DR monoclonal antibodies. Functional and cytokine release assays were carried out using [3H] thymidine uptake test and commercially available ELISA kits for the determination of IL-12, IL-10, IFN-γ and IL-4, respectively. We found that MACS-DCs were more homogenous and the yield and viability were fairly higher than Flask-DCs. MACS-DCs expressed higher levels of CD83 and HLA-DR as well as CD14 compared to the Flask-DCs. Induction of T cell proliferative responses were higher in Flask-DCs and also they elicited higher levels of IL-12: IL-10 and IFN-γ: IL-4 ratios in cytokine generation assays. MACS method was superior for mass production of viable homogenous and fully mature DCs but their cytokine profile had the potential to polarize the immune system toward Th2 type immune response.


Automated prediction of the Thoracolumbar Injury Classification and Severity Score from CT using a novel deep learning algorithm.
Damage to the thoracolumbar spine can confer significant morbidity and mortality. The Thoracolumbar Injury Classification and Severity Score (TLICS) is used to categorize injuries and determine patients at risk of spinal instability for whom surgical intervention is warranted. However, calculating this score can constitute a bottleneck in triaging and treating patients, as it relies on multiple imaging studies and a neurological examination. Therefore, the authors sought to develop and validate a deep learning model that can automatically categorize vertebral morphology and determine posterior ligamentous complex (PLC) integrity, two critical features of TLICS, using only CT scans. All patients who underwent neurosurgical consultation for traumatic spine injury or degenerative pathology resulting in spine injury at a single tertiary center from January 2018 to December 2019 were retrospectively evaluated for inclusion. The morphology of injury and integrity of the PLC were categorized on CT scans. A state-of-the-art object detection region-based convolutional neural network (R-CNN), Faster R-CNN, was leveraged to predict both vertebral locations and the corresponding TLICS. The network was trained with patient CT scans, manually labeled vertebral bounding boxes, TLICS morphology, and PLC annotations, thus allowing the model to output the location of vertebrae, categorize their morphology, and determine the status of PLC integrity. A total of 111 patients were included (mean ± SD age 62 ± 20 years) with a total of 129 separate injury classifications. Vertebral localization and PLC integrity classification achieved Dice scores of 0.92 and 0.88, respectively. Binary classification between noninjured and injured morphological scores demonstrated 95.1% accuracy. TLICS morphology accuracy, the true positive rate, and positive injury mismatch classification rate were 86.3%, 76.2%, and 22.7%, respectively. Classification accuracy between no injury and suspected PLC injury was 86.8%, while true positive, false negative, and false positive rates were 90.0%, 10.0%, and 21.8%, respectively. In this study, the authors demonstrate a novel deep learning method to automatically predict injury morphology and PLC disruption with high accuracy. This model may streamline and improve diagnostic decision support for patients with thoracolumbar spinal trauma.


Phosphorylase kinase phosphorylation of skeletal-muscle troponin T.
Rabbit skeletal-muscle troponin T was phosphorylated by a standard preparation of phosphorylase kinase [Cohen (1973) Eur. J. Biochem. 34, 1--14] and by fractions obtained after chromatography of phosphorylase kinase on phosphocellulose. The original preparation of phosphorylase kinase phosphorylated at least two sites, one of which was serine-1. The second and probably the third sites were presumably located in the peptide flanked by amino-acid residues 147 and 161 of troponin T. Fractions of phosphorylase kinase was adsorbed on phosphocellulose phosphorylated only the second site. Tightly adsorbed fractions possessed high troponin T kinase and phosvitin kinase activities and phosphorylated only serine-1 of troponin T. The results suggest that standard preparations of phosphorylase kinase are contaminated by troponin T kinase, which can phosphorylate serine-1 of troponin T.


Complement-mediated thrombotic microangiopathy as a link between endothelial damage and steroid-refractory GVHD.
Transplant-associated thrombotic microangiopathy (TA-TMA), a complication of hematopoietic cell transplant (HCT), is associated with significant morbidity and mortality. The pathophysiology and overlap of TA-TMA with other posttransplant complications such as graft-versus-host disease (GVHD) is poorly understood. We retrospectively identified cases of TA-TMA among patients with grade 3/4 gastrointestinal (GI) GVHD, reviewed intestinal biopsy specimens, and performed correlative testing of biomarkers associated with TA-TMA. TA-TMA was more common in patients with steroid-refractory GVHD compared with steroid-responsive GVHD (79.3% vs 42.1%; P = .001). Among patients surviving 100 days post-HCT, 1-year survival from day 100 was significantly better for patients who had not developed TA-TMA in the first 100 days (69.5% vs 36.7%; P < .001). Only 1 of 7 proposed TA-TMA histology criteria (mucosal hemorrhage) differed significantly based on GVHD steroid response. In multivariable modeling, steroid-refractory GVHD was a risk factor for development of TA-TMA (hazard ratio, 3.09; 95% confidence interval, 1.68-5.67; P < .001). There were no differences in complement activation at GVHD onset; however, 2 to 6 weeks later, patients with TA-TMA had higher levels of BBPlus and C5b-9, markers of alternative and terminal pathway activation (BBPlus: median, 600 vs 209.3 ng/mL; P = .0045) (C5b-9: median, 425.9 vs 258.4 ng/mL; P = .029). TA-TMA is associated with poor overall survival (OS) following HCT and may be detected early by histologic findings and may be differentiated from GVHD by measurement of alternative and terminal complement pathway activation. It is unknown whether treatment of TA-TMA will improve survival in steroid-refractory GVHD.


Requiem.
"In never considering neuromuscular disease to be untreatable, Yves Rideau has found ways to ameliorate every aspect of these conditions. His work has resulted in immeasurably enhancing the quality of life of his patients". This dedication included into the Guide to Evaluation in Management of Neuromuscular Diseases, 1999, made by a coworker who studied in Poitiers from 1981 to 1983, summarizes the content of this paper.


Invasive Streptococcus pyogenes Infections in <3-Month-Old Infants in France: Clinical and Laboratory Features.
Few data are available on invasive group A Streptococcus (GAS) infections (IGASIs) in infants. We described initial clinical and laboratory features and outcomes of <3-month-old infants hospitalized for an IGASI between 2007 and 2016 in France. Patients were identified from the French National Reference Centre for streptococci. IGASI was defined by the isolation of GAS from blood cultures or from other usually sterile sites. Data collection was performed by assessing the patients' hospitalization reports. Twenty-six patients (15 males; 57.7%) were included. Among 19 cases with available data, 14 (73.7%) were household contacts of a GAS infection, reaching 8/9 (88.9%) in neonates. The diagnoses were bacteremia (n = 18; 69.2%), pleural effusion or pneumonia (n = 6; 23.1%), meningitis with brain abscess (n = 1; 3.8%), and septic arthritis (n = 1; 3.8%). Fever (n = 10; 38.5%), hemodynamic disorders (n = 11; 42.3%), respiratory disorders (n = 7; 26.9%), thrombocytopenia (n = 7; 26.9%), and neutropenia (n = 5; 19.2%) were frequently observed. The main emm-genotype was emm-1 (n = 8; 30.8%). Thirteen (50.0%) infants have been admitted to the intensive care unit, and two (7.7%) died. Respiratory disorders, high C-reactive protein level, and the need for transfusion were significantly associated with severity. IGASI remains uncommon in <3-month-old children but leads to a high morbidity. Whether an antibiotic prophylaxis for contact neonates of a patient with GAS infection decreases the risk of infection remains to be determined.


Feature Extraction for Incomplete Data Via Low-Rank Tensor Decomposition With Feature Regularization.
Multidimensional data (i.e., tensors) with missing entries are common in practice. Extracting features from incomplete tensors is an important yet challenging problem in many fields such as machine learning, pattern recognition, and computer vision. Although the missing entries can be recovered by tensor completion techniques, these completion methods focus only on missing data estimation instead of effective feature extraction. To the best of our knowledge, the problem of feature extraction from incomplete tensors has yet to be well explored in the literature. In this paper, we therefore tackle this problem within the unsupervised learning environment. Specifically, we incorporate low-rank tensor decomposition with feature variance maximization (TDVM) in a unified framework. Based on orthogonal Tucker and CP decompositions, we design two TDVM methods, TDVM-Tucker and TDVM-CP, to learn low-dimensional features viewing the core tensors of the Tucker model as features and viewing the weight vectors of the CP model as features. TDVM explores the relationship among data samples via maximizing feature variance and simultaneously estimates the missing entries via low-rank Tucker/CP approximation, leading to informative features extracted directly from observed entries. Furthermore, we generalize the proposed methods by formulating a general model that incorporates feature regularization into low-rank tensor approximation. In addition, we develop a joint optimization scheme to solve the proposed methods by integrating the alternating direction method of multipliers with the block coordinate descent method. Finally, we evaluate our methods on six real-world image and video data sets under a newly designed multiblock missing setting. The extracted features are evaluated in face recognition, object/action classification, and face/gait clustering. Experimental results demonstrate the superior performance of the proposed methods compared with the state-of-the-art approaches.


Effects of alpha-lipoic acid and dihydrolipoic acid on expression of proto-oncogene c-fos.
The transcription factor AP-1 is an important human mediator of the cellular response to serum, growth factors, and phorbol esters such as 12-O-tetradecanoyl-phorbol-13 acetate (TPA). The AP-1 complex consists of distinct protein heterodimers encoded by the proto-oncogene c-fos and c-jun mRNA whose gene expression can be induced by TPA, cyclic AMP and growth factors. Recent findings suggest an involvement of reactive oxygen species in the pathway of TPA and protein kinase C leading to expression of c-fos and c-jun mRNA. To investigate the role of reactive oxygen species we studied the effects of alpha-lipoic acid and dihydrolipoic acid (natural thiol antioxidants) on the expression of c-fos mRNA in human Jurkat T cells. When cells were preincubated with dihydrolipoic acid (0.2 mM) the expression of c-fos mRNA was suppressed at 30 min after stimulation of TPA (0.5 microM) whereas in the case of preincubation of alpha-lipoic acid (0.2 microM), the expression was enhanced at 30 min. These studies support the idea that superoxide anion radical plays a role in the expression of c-fos mRNA.


Isolation and molecular characterization of Acanthamoeba genotypes in recreational and domestic water sources from Jamaica, West Indies.
Free living amoebae (FLA) are amphizoic protozoa that are ubiquitous in nature. Infection with FLA may result in neurological, ocular and skin infections. Exposure to Acanthamoeba occurs frequently through water contact and knowledge of the presence of the organisms in water sources is important in understanding transmission dynamics. The distribution of Acanthamoeba was studied in recreational and domestic water samples collected from across Jamaica. Morphological assessment and polymerase chain reaction revealed Acanthamoeba spp. isolates in 50.6% (42/83) and 17.3% (14/81) of recreational and domestic water, respectively. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T3, T4, T5, T10 and T11 corresponding to Acanthamoeba spp: A. griffini, A. triangularis, A. lenticulata, A. culbertsoni and A. hatchetti. Moreover, T4 was the most frequently isolated genotype in both recreational and domestic water. Thermotolerance and osmotolerance assays indicated that most isolates were potentially pathogenic. This is the first report of T3 and T10 genotypes in the Caribbean and the first report of these Acanthamoeba spp. in Jamaican waters. The study shows that there is potential risk of infection to contact wearers who practise poor lens care. Further, Acanthamoeba should be considered as a cause of neurological infections in Jamaica.


Flavobacterium verecundum sp. nov., isolated from a freshwater river.
Strain TTM-46T, isolated from a water sample taken from the Caohu River in Taiwan, was characterized using a polyphasic taxonomy approach. Cells of TTM-46T were Gram-staining-negative, strictly aerobic, motile by gliding, rod-shaped, surrounded by a thick capsule and formedyellowish-green colored colonies. Growth occurred at 20-30 °C (optimum, 30 °C), at pH 6-8 (optimum, pH 7) and with 0-0.3 % NaCl (optimum, 0 % w/v). Phylogenetic analyses based on 16 S rRNA gene sequences showed that TTM-46T represented a member of the genus Flavobacterium and was most closely related to Flavobacterium columnare IFO 15943T and Flavobacterium brevivitae TTM-43T with a sequence similarity of 96.9 % for both. TTM-46T contained iso-C15 : 0, summed feature 9 (iso-C17 : 1ω9c and/or 10-methyl C16 : 0), iso-C17 : 0 3-OH, iso-C15 : 1 G, iso-C15 : 0 3-OH and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) as the predominant fatty acids. The major isoprenoid quinone was MK-6. The polar lipid profile consisted of phosphatidylethanolamine, four uncharacterized aminophospholipids and five uncharacterized phospholipids. The major polyamine was homospermidine. The genomic DNA G+C content of TTM-46T was 39 mol%. On the basis of the phylogenetic inference and phenotypic data, TTM-46T was recognized as a representative of a novel species within the genus Flavobacterium. The name Flavobacterium verecundum sp. nov. is proposed, with TTM-46T (=BCRC 80914T=LMG 29005T=KCTC 42745T) as the type strain.


Histologic Evidence of New Collagen Formulation Using Platelet Rich Plasma in Skin Rejuvenation: A Prospective Controlled Clinical Study.
Platelet-rich plasma (PRP) is an autologous concentration of human platelets contained in a small volume of plasma and has recently been shown to accelerate rejuvenate aging skin by various growth factors and cell adhesion molecules. This study was conducted to evaluate the efficacy and safety of intradermal injection of PRP in the human facial rejuvenation. This study was a prospective, single-center, single-dose, open-label, non-randomized controlled clinical study. PRP injected to the upper site of this right infra-auricular area and all face. Saline was injected to the left infra-auricular area. Histopathological examinations were performed before PRP treatment, 28 days after the PRP, and saline (control) treatments. Twenty women ranging in age from 40 to 49 years (mean age, 43.65±2.43 years) were enrolled in the study. The mean optical densities (MODs) of collagen in the pre-treatment, control, and PRP-treated area were measured. They were 539±93.2, 787±134.15, 1,019±178, respectively. In the MOD of PRP, 89.05 percent improvement was found when MOD of PRP was compared with MOD of pre-treatment. The mean MOD of collagen fibers was clearly highest on the PRP side (p<0.001). The PRP-to-saline improvement ratio (89.05% to 46.01%) was 1.93:1. No serious side effects were detected. PRP increases dermal collagen levels not only by growth factors, but also by skin needling (the mesotherapy technique 'point by point'). PRP application could be considered as an effective (even a single application) and safety procedure for facial skin rejuvenation.


A Comprehensive Molecular Characterization of the Pancreatic Neuroendocrine Tumor Cell Lines BON-1 and QGP-1.
Experimental models of neuroendocrine tumor disease are scarce, with only a few existing neuroendocrine tumor cell lines of pancreatic origin (panNET). Their molecular characterization has so far focused on the neuroendocrine phenotype and cancer-related mutations, while a transcription-based assessment of their developmental origin and malignant potential is lacking. In this study, we performed immunoblotting and qPCR analysis of neuroendocrine, epithelial, developmental endocrine-related genes as well as next-generation sequencing (NGS) analysis of microRNAs (miRs) on three panNET cell lines, BON-1, QGP-1, and NT-3. All three lines displayed a neuroendocrine and epithelial phenotype; however, while insulinoma-derived NT-3 cells preferentially expressed markers of mature functional pancreatic β-cells (i.e., INS, MAFA), both BON-1 and QGP-1 displayed high expression of genes associated with immature or non-functional β/δ-cells genes (i.e., NEUROG3), or pancreatic endocrine progenitors (i.e., FOXA2). NGS-based identification of miRs in BON-1 and QGP-1 cells revealed the presence of all six members of the miR-17-92 cluster, which have been implicated in b-cell function and differentiation, but also have roles in cancer being both oncogenic or tumor suppressive. Notably, both BON-1 and QGP-1 cells expressed several miRs known to be negatively associated with epithelial-mesenchymal transition, invasion or metastasis. Moreover, both cell lines failed to exhibit migratory activity in vitro. Taken together, NT-3 cells resemble mature functional β-cells, while both BON-1 and QGP-1 are more similar to immature/non-functional pancreatic β/δ-cells or pancreatic endocrine progenitors. Based on the recent identification of three transcriptional subtypes in panNETs, NT-3 cells resemble the "islet/insulinoma tumors" (IT) subtype, while BON-1 and QGP-1 cells were tentatively classified as "metastasis-like/primary" (MLP). Our results provide a comprehensive characterization of three panNET cell lines and demonstrate their relevance as neuroendocrine tumor models.


Individual and maternal determinants of self-reported dental health among Turkish school children aged 10-12 years.
To assess the influence of maternal and individual characteristics on self-reported dental health of Turkish school children aged 10-12 years with different socio-economic backgrounds. A cross-sectional study of children aged 10 to 12 (n = 611) using paired matches of self-administered questionnaires for children and their mothers. Clinical examinations based on World Health Organization criteria were conduced. The participation rate was 97% (n = 591) for the children, 87% (n = 533) for the mothers, and 95% (n = 584) for the clinical examinations. Multiple linear regression, descriptive statistics, Spearman correlation coefficient and chi-square test were applied. Private school children's mothers were more likely to have had higher education (95% at least high school) than public school children's mothers (11%); they reported better dental health (above average) than did mothers of public school children (p = 0.001). Among all mothers, those with above average self-reported dental health reported more regular dental visits than did those with below average scores (p = 0.001). Frequency of regular dental visits and toothbrushing among children attending public schools (5%, 65%) were lower than those attending private school (43%, 79%), (p = 0.001). The mean DMFS was negatively correlated with self-reported dental health (r(s) = -0.187, p = 0.001). Toothbrushing frequency and school performance were the common factors positively associated with self-reported dental health, among all children. The results emphasize the important role of mothers and their socio-economic background in enhancement of children's dental health. Their active role in conjunction with the potential of self-assessment provides a good basis for establishing and improving self-care among children, in developing countries in particular.


Congenital hypothyroidism in a kitten resulting in decreased IGF-I concentration and abnormal liver function tests.
A 7-month-old male kitten was presented with chronic constipation and retarded growth. Clinical examination revealed disproportional dwarfism with mild skeletal abnormalities and a palpable thyroid gland. The presumptive diagnosis of congenital hypothyroidism was confirmed by low serum total thyroxine (tT(4)) concentration prior to and after the administration of thyroid stimulation hormone (TSH), increased endogenous TSH concentration and abnormal thyroid scintigraphic scan. The kitten had abnormal liver function tests and decreased insulin-like growth factor 1 (IGF-1) concentration, both of which returned to normal in correspondence with an improvement of the clinical signs after 6 weeks of thyroxine therapy. Congenital hypothyroidism is a rare disease that may present with considerable variation in clinical manifestation. In cases in which clinical signs are ambiguous, disorders such as portosystemic shunt and hyposomatotropism have to be taken into account as differential diagnosis. As hypothyroidism may be associated with abnormal liver function tests and low IGF-1 concentrations, test results have to be interpreted carefully.


Phospholipase D2 directly interacts with aldolase via Its PH domain.
Mammalian phospholipase D (PLD) has been implicated in the cellular signal transduction pathways leading to diverse physiological events and known to be regulated by many cellular factors. To identify the proteins that interact with PLD, we performed a protein overlay assay with fractions obtained from the sequential column chromatographic separation of rat brain cytosol using purified PLD2 as a probe. A protein of molecular mass 40 kDa, which was detected by anti-PLD antibody with overlaying of the purified PLD2, is shown to be aldolase C by peptide-mass fingerprinting with matrix-assisted laser desorption/ionization-time-of flight mass spectrometry (MALDI-TOF-MS). Aldolase A also showed similar binding properties as aldolase C and was co-immunoprecipitated with PLD2 in COS-7 cells overexpressing PLD2 and aldolase A. The PH domain corresponding to amino acids 201-310 of PLD2 was necessary for the interaction observed in vitro, and aldolase A was found to interact with the PH domain of PLD2 specifically, but not with other PH domains. PLD2 activity was inhibited by the presence of purified aldolase A in a dose-dependent manner, and the inhibition by 50% was observed by the addition of less than micromolar aldolase A. Moreover, the inclusion of the aldolase metabolites fructose 1,6-bisphosphate (F-1,6-P) or glyceraldehyde 3-phosphate (G-3-P) resulted in an enhanced interaction between PLD2 and aldolase A with a concomitant increase in the potential ability of aldolase A to inhibit PLD2, which suggests the existence of a possible regulation of the interaction by the change of intracellular concentrations of glycolytic metabolites.


Morphological and functional abnormalities of hippocampus in APC1638T/1638T mice.
In the present study, we examined morphology and function of hippocampus in the APC1638T/1638T mouse. Expression levels of the APC mRNA and protein were both identical in the hippocampus of the APC+/+ and APC1638T/1638T mice. The dentate gyrus of the APC1638T/1638T hippocampus was thicker, and has more densely-populated granule cells in the APC1638T/1638T mouse hippocampus. Immunoelectron microscopy revealed co-localization of APC with alpha-amino-3- hydroxy-5-methyl- isoxazole-4-propionate receptor (AMPA-R) and with PSD-95 at post-synapse in the APC+/+ hippocampus, while APC1638T was co-localized with neither AMPA-R nor PSD-95 in the APC1638T/1638T hippocampus. By immunoprecipitation assay, full-length APC expressed in the APC +/+ mouse was co-immunoprecipitated with AMPA-R and PSD-95. In contrast, APC1638T expressed in the APC1638T/1638T mouse was not co-immunoprecipitated with AMPA-R and PSD-95. In the hippocampal CA1 region of the APC1638T/1638T mouse, c-Fos expression after electric foot shock was decreased compared with the APC+/+ mouse. The present study showed some abnormalities on morphology of the hippocampus caused by a truncated APC (APC1638T). Also, our findings suggest that failure in APC binding to AMPA-R and PSD-95 may bring about less activities of hippocampal neurons in the APC1638T/1638T mouse.


Serologic evidence in commercial chicken and turkey flocks of infection with reticuloendotheliosis virus.
A serologic survey has documented probable infection with reticuloendotheliosis (RE) virus in 21.0% of 101 layer flocks, 23.5% of 85 broiler and broiler-breeder flocks, 2.3% of 43 backyard chicken flocks, and 4.8% of 125 turkey production and breeder flocks. However, no infection was detected in 72 grandparent lines of chicken breeding stocks representing meat-type and layer strains. The existence of natural infection was further supported by isolation of RE virus from one experimental chicken flock and two commercial turkey flocks. This study supports earlier but subsequently discounted data by Aulisio and Shelokov that exposure to RE virus occurs commonly among commercial chickens in the United States, as has also been reported in other countries.


Simultaneous photocatalytic degradation of ibuprofen and H2 evolution over Au/sheaf-like TiO2 mesocrystals.
Considerable effort has been devoted to the efficient degradation of pharmaceuticals and personal care products (PPCPs), while the chemical energy in these processes has been widely overlooked. In this study, we demonstrated the simultaneous hydrogen production and ibuprofen degradation through heterogeneous photocatalysis. By anchoring Au nanoparticles (NPs) on the (101) surface of sheaf-like TiO2 mesocrystals with [001] orientation, efficient charge separation is achieved, which is essential for the photocatalytic redox reactions. XPS analysis showed that the binding energies of Ti 2p and O 1s indicated no shift after Au addition. Peaks observed at 81.8 and 85.5 eV due to Au 4f7/2 and Au 4f5/2 of metallic gold on the surface of Au/meso-TiO2, confirmed the formation of Au NPs. The as-synthesized anatase TiO2 mesocrystals are composed of small nanocrystals with a size of 8 nm and exhibit the uniform sheaf-like morphology along [001] orientation. As expected, the 1 wt% Au/TiO2 mesocrystals shows the largest photocurrent density, highest H2-evolution rate, and fastest photodegradation rate of ibuprofen under simulated sunlight irradiation among all the studied catalyst. Furthermore, the effect of solution pH, common anions (Cl-, NO3-, and SO42-) and cations (Na+, K+, and Ca2+) on photocatalytic H2 evolution and degradation of ibuprofen were individually investigated and discussed. A mechanism for the simultaneous photocatalytic hydrogen generation and degradation of ibuprofen has also been proposed. This work opens up new opportunities for the development of energy efficient techniques for PPCPs degradation.


The epidemiology of diabetic neuropathy. Diabetic Cardiovascular Autonomic Neuropathy Multicenter Study Group.
Although neuropathy has long been recognized as a complication of diabetes, the impact of this condition has not been adequately established. The prevalence of diabetic neuropathy is virtually unknown because the published studies differ considerably with regard to definition, method of assessment, and patient selection. Furthermore, the determination of prevalence has been hampered by the fact that there is no generally accepted classification of the variety of manifestations of diabetic neuropathy. The introduction of new sensitive diagnostic methods aids in the detection of less severe stages of neuropathy, as compared with clinically based assessment, and renders the disease more prevalent. The prevalence of diabetic neuropathy in the few reported population-based studies was approximately 30%. We have evaluated the prevalence of cardiovascular autonomic neuropathy in a group of approximately 1000 diabetic patients randomly included from 21 hospitals in Germany, Austria, and Switzerland. The results of this study and those of a prospective study on the natural history of neural dysfunction during the first 5 years after diagnosis of type 1 diabetes will be presented.


Barriers and facilitators to goals of care conversations with Veteran residents of community nursing homes.
Despite evidence that structured goals of care conversations (GoCCs) and documentation of life-sustaining treatment (LST) preferences improve the delivery of goal-concordant care for seriously ill patients, rates of completion remain low among nursing home residents. The Preferences Elicited and Respected for Seriously Ill Veterans through Enhanced Decision-Making (PERSIVED) program aims to improve the consistent documentation of LST preferences among Veterans receiving care in veterans affairs (VA)-paid community nursing homes (CNH); however, the barriers and facilitators of completing and documenting GoCCs in this unique context of care have not been described. We conducted semi-structured, qualitative interviews with key stakeholders of the VA CNH programs located at six VA Medical Centers between July 2021 and July 2022. With a rapid approach to analysis, interview transcripts were reduced into memo templates using the Tailored Implementation for Chronic Disease Checklist and coded and analyzed using qualitative data analysis software. The 40 participants consisted of nurses (n = 13), social workers (n = 25), and VA physicians (n = 2). Most participants felt confident about conducting GoCC; however, several barriers were identified. At the staff level, our results indicated inconsistent completion of GoCC and documentation due to a lack of training, confusion about roles and responsibilities, and challenging communication within the VA as well as with CNH. At the organizational level, there was a lack of standardization across sites for how LST preferences were documented. At the patient level, we found key barriers related to patient and family readiness and issues finding surrogate decision makers. While COVID-19 brought end-of-life issues to the forefront, lockdowns hindered communication about the goals of care. Findings from this pre-implementation evaluation revealed multi-level barriers in conducting and documenting GoCCs with Veterans receiving VA-paid CNH care, as well as several facilitators that can be used to inform strategies for improvement.


Comparison of MPTP-induced changes in spontaneous neuronal discharge in the internal pallidal segment and in the substantia nigra pars reticulata in primates.
The basal ganglia are currently viewed as components of segregated corticosubcortical reentrant circuits. One of these circuits, the "motor" circuit, is critically involved in the development of parkinsonian motor signs. Current pathophysiologic models postulate that parkinsonism is associated with increased activity in the basal ganglia output nuclei. The neuronal activity in the motor portion of one of these output nuclei, the internal segment of the globus pallidus (GPi), has been characterized in detail in intact and parkinsonian animals, but the neuronal activity in the second major basal ganglia output nucleus, the substantia nigra pars reticulata (SNr), has received far less attention. This study in primates represents a comparison of the effects of parkinsonism, induced by injections of the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), on the neuronal discharge in the GPi and SNr. These electrophysiologic recording experiments were carried out in three African green and two rhesus monkeys. One hundred and twenty-four neurons were recorded in the GPi before treatment with MPTP, and 93 neurons thereafter. In the SNr, 55 cells were recorded before treatment with MPTP, and 41 cells thereafter. MPTP induced a non-significant increase in the average discharge rate and a significant decrease in the median interspike interval length (ISI) in the GPi (by 13%), whereas no changes were detected in either parameter in the SNr. The average ISI distributions were markedly asymmetric in both structures, and could be modeled by a logarithmic normal distribution. With the MPTP treatment, the mode of the ISI distribution fell by 24% in the GPi (P< or =0.01), whereas it did not change significantly in the SNr. An algorithm that detects burst discharges in the raw ISI data (based on the method by Legendy and Salcman) detected a significant increase in the proportion of action potentials that participated in bursts of discharge in both structures (increase by 257% in the GPi, and by 67% in the SNr). Power spectral and autocorrelation analysis revealed that treatment with MPTP increased the proportion of cells with oscillatory burst patterns at 3-8 Hz in both structures (from 0.8% to 27% of all neurons in the GPi, and from none to 10% in the SNr). The results show that neuronal discharge in the SNr is affected in parkinsonism, but that the changes in the SNr are less pronounced then those seen in the GPi.


Aging and extrapyramidal function.
Measurements on human brain samples of some enzymes concerned with neurotransmitter synthesis suggest serious losses with age. The most severe loss found was that in striatal tyrosine hydroxylase activity, the rate-controlling enzyme in the synthesis of dopamine. Cell counts in the substantia nigra where this dopaminergic tract originates suggest that the decrease in enzyme activity is partly due to cell loss, but must largely reflect decreased activity of residual cells. It is possible that this loss may account for some of the difficulties in movement seen in aged individuals and that it might be less if pigment formation in these cells could be inhibited.


[Cohort study on risk factors for subarachnoid hemorrhage among Japanese men and women].
To examine the risk factors for subarachnoid hemorrhage, for few prospective data have been available on risk factors of subarachnoid hemorrhage among Japanese. A 9.4-year cohort study was conducted on 12,372 men and women, aged 40-69 years, free of history of stroke in six communities in Japan. During the follow-up, seventy-one incidents of subarachnoid hemorrhage occurred. After adjusting for age, serum total cholesterol, body mass index, history of diabetes mellitus, blood pressure category and drinking category, current smokers had a significantly increased risk of subarachnoid hemorrhage compared with 'never-smokers': multivariate relative risk (RR) [95% confidence interval (95% CI)] = 3.1(1.4-6.9) for women, 1.5(0.4-5.3) for men and 2.7(1.3-5.5)for men and women. Hypertensives had a significantly increased risk of subarachnoid hemorrhage compared with normotensives: multivariate RR(95% CI) = 3.1(1.6-6.0) for women, 4.3(1.5-12.0) for men and 3.4(2.0-5.9) for men and women. For men, heavy drinkers (> = 69 g per day ethanol), had a significantly increased risk of subarachnoid hemorrhage compared with 'never-drinkers': multivariate RR(95% CI) = 4.3(1.1-16.8). For women, the relationship between heavy drinking and risk was not examined due to the small number of heavy drinkers (n = 13). Multivariate RR(95% CI) for men who smoked currently and drank heavily compared with men who did not smoke currently nor drink heavily was 6.0(1.8-20.1). Multivariate RR(95% CI) for hypertensive men who drank heavily compared with normotensive or borderline hypertensive men who did not drink heavily was 13.0(3.9-43.9). Multivariate RR(95% CI) for hypertensive men and women who smoked currently compared with normotensive or borderline hypertensive men and women who did not smoke currently was 6.1(1.3-28.7) for men and 6.3(1.4-28.0) for women. Furthermore, the combination of smoking, hypertension and heavy drinking raised the risk of subarachnoid hemorrhage substantially: multivariate RR(95% CI) = 17.5(3.4-90.2). Smoking and hypertension appeared to increase the risk of subarachnoid hemorrhage. For men, although the association of smoking with the risk of subarachnoid hemorrhage was weak compared with women, the combination of smoking, hypertension and heavy drinking increased the risk substantially. For women, smoking appeared to be an independent risk of subarachnoid hemorrhage, and the combination of smoking and hypertension increased the risk in additive fashion.


paraSBOLv: a foundation for standard-compliant genetic design visualization tools.
Diagrams constructed from standardized glyphs are central to communicating complex design information in many engineering fields. For example, circuit diagrams are commonplace in electronics and allow for a suitable abstraction of the physical system that helps support the design process. With the development of the Synthetic Biology Open Language Visual (SBOLv), bioengineers are now positioned to better describe and share their biological designs visually. However, the development of computational tools to support the creation of these diagrams is currently hampered by an excessive burden in maintenance due to the large and expanding number of glyphs present in the standard. Here, we present a Python package called paraSBOLv that enables access to the full suite of SBOLv glyphs through the use of machine-readable parametric glyph definitions. These greatly simplify the rendering process while allowing extensive customization of the resulting diagrams. We demonstrate how the adoption of paraSBOLv can accelerate the development of highly specialized biodesign visualization tools or even form the basis for more complex software by removing the burden of maintaining glyph-specific rendering code. Looking forward, we suggest that incorporation of machine-readable parametric glyph definitions into the SBOLv standard could further simplify the development of tools to produce standard-compliant diagrams and the integration of visual standards across fields.


Effectiveness of surgery for adults with hallux valgus deformity: a systematic review.
Hallux valgus (HV) is a common foot deformity. In severe stages of this condition, surgery is often necessary. Currently, there is no systematic review comparing the effectiveness of surgery over conservative treatment. The objective of this review was to establish the effectiveness of surgery compared to conservatory management for adults with HV. The current review included adults (18 years or over) with HV deformity, excluding adults with neurological problems causing foot deformities, for example, cerebral palsy, neuropathy, stroke and multiple sclerosis. The review included any type of HV surgery compared to no surgery, conservative treatment or different types of HV surgeries. The primary outcome was gait measurement, and secondary outcomes included quality of life, patient satisfaction, pain assessed using any validated assessment tool and adverse events. The review included randomized controlled trials. The search strategy aimed to find both published and unpublished studies. A three-step search strategy was utilized in 16 databases without language and date limitations. Papers selected for retrieval were assessed by two independent reviewers for methodological validity prior to inclusion in the review using standardized critical appraisal instruments developed by the Joanna Briggs Institute (JBI). Data were extracted from papers included in the review using the standardized data extraction tool developed by the JBI. Quantitative data were, where possible, pooled in statistical meta-analysis using RevMan5 (Copenhagen: The Nordic Cochrane Centre, Cochrane). Effect sizes expressed as risk ratio (for categorical data) and mean differences (MD) or standardized MD (for continuous data) and their 95% confidence intervals were calculated for analysis. Where statistical pooling was not possible, the findings have been presented in narrative form. Searching identified 2412 citations. After removal of duplicates, paper retrieval and critical appraisal, 25 studies were included in the review. The included trials were of medium-to-poor quality.Twenty-four trials compared the effectiveness of different types of surgeries. Meta-analysis revealed no difference in level of pain between distal chevron-type osteotomy and other surgical procedures (standard mean difference [SMD] 0.02, 95% confidence interval [CI] -0.24 to 0.28). One single trial reported that distal chevron osteotomy is more effective than Lindgren osteotomy in terms of walking speed (MD -0.24, 95% CI -0.43 to -0.05).One medium quality trial assessed the effectiveness of HV surgery compared to conservative or no treatment. This trial showed that surgery, specifically distal chevron osteotomy of the first metatarsal, is a more effective procedure for pain compared to conservative treatment (MD -15.00, 95% CI -22.79 to -7.21) and also no treatment in the first year (MD -18.00, 95% CI -25.62 to -10.38). The current systematic review showed that differences between various types of surgical procedures, specifically osteotomies of the first metatarsal on clinical outcomes, are minimal. There is evidence from one study, that surgery, specifically distal chevron osteotomy of the first metatarsal is a more effective procedure than conservative or no treatment in reducing pain in the first year following surgery. However, this systematic review has identified that there is a lack of high-quality studies comparing similar types of HV treatments that assess the same outcomes.


A heat shock protein 90 inhibitor reduces oncoprotein expression and induces cell death in heterogeneous glioblastoma cells with EGFR, PDGFRA, CDK4, and NF1 aberrations.
Glioblastoma (GBM) is a highly malignant brain tumor. After treatment with the first-line drug temozolomide, only 50% of patients are responsive. Recent literature shows that the difficulty in treating GBM is mainly due to the heterogeneity of its four major cellular states, which are characterized by differences in EGFR, PDGFRA, CDK4, and NF1. Therefore, development of a multitarget drug is a potential strategy for treating heterogeneous GBM. In this study, the antitumor ability of a potent heat shock protein 90 inhibitor, NVP-AUY922 (AUY922), was evaluated in GBM cell lines (U-87 MG and T98G cells) and patient-derived GBM cell lines [P#5 and P#5 temozolomide-resistant (TMZ-R) cells]. We found that AUY922 significantly reduced cell viability and colony formation in four GBM cell lines. AUY922 also significantly induced apoptosis by increasing PARP1 cleavage and the number of annexin V-positive cells. The autophagy indicators as MAP1LC3B cleavage and MAP1LC3B puncta were increased after AUY922 treatment. AUY922-induced cell death could be partially reversed by pharmacological inhibition of either apoptotic inhibitor or autophagy inhibitor. Moreover, AUY922 reduced the mRNA and protein expressions of EGFR, PDGFRA, CDK4, and NF1, which contribute to the four cellular state subtypes in GBM cells. In addition, the downstream signaling proteins of these four proteins, AKT/p-AKT, MAPK/p-MAPK, and BRAF, were downregulated after AUY922 treatment. Taken together, AUY922 led to GBM cell death via apoptosis and autophagy, and reduced the mRNA and protein expression of EGFR, PDGFRA, CDK4, and NF1in heterogeneous GBM cells.


On surface-initiated atom transfer radical polymerization using diazonium chemistry to introduce the initiator layer.
This work features the controllability of surface-initiated atom transfer radical polymerization (SI-ATRP) of methyl methacrylate, initiated by a multilayered 2-bromoisobutyryl moiety formed via diazonium chemistry. The thickness as a function of polymerization time has been studied by varying different parameters such as the bromine content of the initiator layer, polarity of reaction medium, ligand type (L), and the ratio of activator (Cu(I)) to deactivator (Cu(II)) in order to ascertain the controllability of the SI-ATRP process. The variation of thickness versus surface concentration of bromine shows a gradual transition from mushroom to brush-type conformation of the surface anchored chains in both polar and nonpolar reaction medium. Interestingly, it is revealed that very thick polymer brushes, on the order of 1 μm, can be obtained at high bromine content of the initiator layer in toluene. The initial polymerization rate and the overall final thickness are higher in the case of nonpolar solvent (toluene) compared to polar medium (acetonitrile or N,N-dimethylformamide). The ligand affects the initial rate of polymerization, which correlates with the redox potentials of the pertinent Cu(II)/Cu(I) complexes (L = Me(6)TREN, PMDETA, and BIPY). It is also observed that the ability of polymer brushes to reinitiate depends on the initial thickness and the solvent used for generating it.


A de-interlacing algorithm using Markov random field model.
In this paper, a motion-compensated de-interlacing algorithm using the Markov random field (MRF) model is proposed. The de-interlacing problem is formulated as a maximum a posteriori (MAP) MRF problem. The MAP solution is the one that minimizes an energy function, which imposes discontinuity-adaptive smoothness (DAS) spatial constraint on the de-interlaced frame. The edge direction information, which is used to formulate the DAS constraint, is implicitly indicated by weight vectors (weights for 16 digitized directions). Generally, large weights are assigned to along-edge directions and relatively small weights are assigned to across-edge directions. As a local statistical-based method, the proposed weighting method should be more robust than traditional edge-directed interpolation methods in deciding local edge directions. The proposed algorithm is implemented by an iterative optimization process, which guarantees convergence. However, a global optimal solution is not guaranteed due to computational complexity concern. Simulation results compare the proposed algorithm to other motion compensated de-interlacing algorithms. Significant improvements of de-interlaced edges are observed.


Pelvic and lower extremity injuries in Homer's Iliad: a review of the literature.
The Iliad, composed approximately in the middle of the eighth century bc, constitutes the leading and oldest known example of heroic epic. The Homeric epic presents the conflicts that took place during the last year of the 10-year lasting Trojan War, offering a realistic description of battle wounds. We studied the text of The Iliad in ancient Greek and in the translations in modern Greek and English and searched for all recorded injuries to the pelvis and lower extremities. A total of 16 traumatic injuries of pelvis and lower extremities were described, including 7 fatal wounds, while in 9 cases, the outcome was unknown.The Iliad remains the oldest record of Greek medicine and a unique source of surgical history. To study the vividly reported events is a great experience, particularly for a surgeon.


Development and characterization of a magnetic bead-quantum dot nanoparticles based assay capable of Escherichia coli O157:H7 quantification.
The development and characterization of a magnetic bead (MB)-quantum dot (QD) nanoparticles based assay capable of quantifying pathogenic bacteria is presented here. The MB-QD assay operates by having a capturing probe DNA selectively linked to the signaling probe DNA via the target genomic DNA (gDNA) during DNA hybridization. The signaling probe DNA is labeled with fluorescent QD(565) which serves as a reporter. The capturing probe DNA is conjugated simultaneously to a MB and another QD(655), which serve as a carrier and an internal standard, respectively. Successfully captured target gDNA is separated using a magnetic field and is quantified via a spectrofluorometer. The use of QDs (i.e., QD(565)/QD(655)) as both a fluorescence label and an internal standard increased the sensitivity of the assay. The passivation effect and the molar ratio between QD and DNA were optimized. The MB-QD assay demonstrated a detection limit of 890 zeptomolar (i.e., 10(-21) mol L(-1)) concentration for the linear single stranded DNA (ssDNA). It also demonstrated a detection limit of 87 gene copies for double stranded DNA (dsDNA) eaeA gene extracted from pure Escherichia coli (E. coli) O157:H7 culture. Its corresponding dynamic range, sensitivity, and selectivity were also presented. Finally, the bacterial gDNA of E. coli O157:H7 was used to highlight the MB-QD assay's ability to detect below the minimum infective dose (i.e., 100 organisms) of E. coli O157:H7 in water environment.


Problematic Internet use and its associations with health-related symptoms and lifestyle habits among rural Japanese adolescents.
There have been concerns about the increase in problematic Internet use (PIU) and its impact on lifestyle habits and health-related symptoms, given the rapid spread of smartphones. This study aimed to clarify PIU prevalence over 3 years in the same area and investigate lifestyle and health-related factors related to PIU among junior high-school students in Japan. Each year during 2014-2016, a survey was conducted with junior high-school students from a rural area of Japan (2014, n = 979; 2015, n = 968; 2016, n = 940). Young's Internet Addiction Test was used to assess participants' PIU. Students scoring 40 or higher on the Internet Addiction Test were classified as showing PIU in this study. The associations between PIU and lifestyle factors (e.g., exercise habits, weekday study time, and sleep time) and health-related symptoms (depressive symptoms and orthostatic dysregulation [OD] symptoms) were studied by logistic regression analyses. Over the 3 years, the prevalence of PIU was 19.9% in 2014, 15.9% in 2015, and 17.7% in 2016 without significant change. PIU was significantly associated with skipping breakfast, having a late bedtime (after midnight), and having OD symptoms among students from all grades. Sleepiness after awakening in the morning, less studying time, and depressive symptoms had significant positive associations with PIU, except among 1st grade junior high-school students. Our results suggest that PIU is related to decreased time spent sleeping, studying, and exercising and increased symptoms of depression and OD. Further investigations are needed to develop preventive measures for PIU.


Optic Nerve Sheath Diameter Correlation with Elevated Intracranial Pressure Determined via Ultrasound.
Background The early detection of elevated intracranial pressure (ICP) can not only prevent mortality but also aid in more aggressive management. Brain computed tomography (CT) is a mainstay modality in detecting elevated ICP, but the feasibility of using brain CTs to detect elevated ICP in critically ill patients is limited, especially for patients who require high levels of inotropic support. The optic nerve sheath is a direct extension of the brain meninges. Therefore, the elevation of ICP is directly transmitted to the sheath. Measuring the optic nerve sheath diameter (ONSD) through ultrasound (US) is a bedside, noninvasive means to detect elevated ICP. The goal of this study was to assess the correlation of ONSD with elevated ICP as measured via US in an intensive care unit (ICU). Methods We conducted a six-month prospective, single-center, observational study of mass effect stroke patients aged 18 to 65 years who had a traumatic brain injury (TBI) and were admitted to the ICU. Patients with chronic hydrocephalus, extensive local orbit trauma, a pre-existing ocular disease affecting the optic nerve and/or orbital cavity, hyperthyroidism with exophthalmos, and facial trauma affecting the orbits and/or eyeballs were excluded. We measured the ONSD at the entry of optic nerve into the globe using two-dimensional (2D) US. Results One hundred patients were included in the study. Forty-nine patients had diffuse cerebral edema detected on CT scan correlating with increased ONSD notable via bedside US. The mean ONSD related to CT-detectable elevated ICP was 0.61 cm. The sensitivity for the ONSD cut-off value of ≥5.8 mm was 94% (95% confidence interval [CI], 84.05% to 98.79%), and the specificity was 96.08% (95% CI, 86.7% to 99.52%).The positive predictive value was 92.08% (95% CI, 86.28% to 98.96%), and the negative predictive value was 94.23% (95% CI, 84.47% to 98.00%). Conclusion The greatest accuracy in ONSD was found with a cut-off of >0.58 cm in patients with positive CT brain findings. Therefore, US can be used as an initial screening test when physicians suspect a patient has elevated ICP.


Tumor necrosis factor inhibitors in patients with Takayasu arteritis: experience from a referral center with long-term followup.
To report a single-center experience with the use of tumor necrosis factor (TNF) inhibitors in patients with Takayasu arteritis (TA). We retrospectively studied a cohort of patients with refractory TA evaluated at our institution and treated with TNF inhibitors. American College of Rheumatology criteria for TA were used for inclusion. Disease activity was assessed according to the National Institutes of Health criteria. We included 20 patients (19 women, 17 white) with a mean ± SD age of 33 ± 10.2 years and a median disease duration of 15.9 months (interquartile range [IRQ] 2-32.7 months) prior to the use of TNF inhibitors. Before the use of TNF inhibitors, all 20 patients received prednisone. Other medication use included methotrexate (18 patients), azathioprine (5 patients), mycophenolate mofetil (3 patients), and cyclophosphamide (3 patients). Seventeen patients (85%) received infliximab, 2 patients (10%) received adalimumab, and 1 patient (5%) received etanercept. The median duration of treatment with TNF inhibitors was 23.0 months (IQR 8.7-38.9 months). Treatment with TNF inhibitors resulted in disease remission in 18 (90%) of 20 patients and sustained remission in 10 patients (50%). Ten (83%) of 12 patients were able to taper prednisone below 10 mg and 7 patients discontinued prednisone. However, 6 of the 18 patients achieving remission experienced relapse while receiving TNF inhibitors. Eleven patients (55%) discontinued TNF inhibitors for the following reasons: relapse, persistently active disease, lack of corticosteroid-sparing effect, adverse effects (4 patients), and other reasons (4 patients). In this study, treatment with TNF inhibitors induced remission, including sustained remission in patients with refractory TA. However, 33% of patients experienced disease relapse while receiving TNF inhibitors and 20% discontinued treatment because of adverse events.


Evaluation of anionic histological dyes as co-injectable cell markers in pre-implantation mouse embryos.
The enzyme horseradish peroxidase (HRP) is a widely used microinjectable cell marker for studying cell position, lineage, and migration in many kinds of animal embryos. Marked cells are easily identified because they darken when exposed to a chromophore and an HRP substrate such as hydrogen peroxide. This assay, however, requires cytochemical fixation. Thus, when HRP-marked cells need to be identified prior to fixation, visible co-injectants such as dyes and fluorescent substances have been used with HRP. Fluorescent substances have limitations because their excitation could be harmful to the marked cells. Visible but non-fluorescent co-injectants, however, would permit visualization of HRP-marked cells without inflicting such damage. We tested the compatibility of several histological dyes and electrolytic carriers with HRP iontophoresed as a cell marker in 2-cell mouse embryos. The dyes tested were Evans Blue, Cibacron Blue F3GA, Fast Green FCF, and Patent Blue Violet; the electrolytic carriers were KCl, K2SO4, CH3CO2K, and KH2PO4. The combination found most useful was Patent Blue Violet in K2SO4. Survival of embryos incubated to the blastocyst stage following injection with HRP + Patent Blue Violet in K2SO4 at the 2-cell stage was significantly greater than that of embryos injected with any other dye. Although the proportion of embryos undergoing the 8-cell-to-morula transition was somewhat decreased by this treatment, the proportion of embryos reaching the blastocyst stage was comparable to that in the uninjected (control) group. Our results indicate that Patent Blue Violet is a useful, HRP-co-injectable dye for short-term cell marking in preimplantation mouse embryos.


Electrostatic charging and control of droplets in microfluidic devices.
Precharged droplets can facilitate manipulation and control of low-volume liquids in droplet-based microfluidics. In this paper, we demonstrate non-contact electrostatic charging of droplets by polarizing a neutral droplet and splitting it into two oppositely charged daughter droplets in a T-junction microchannel. We performed numerical simulation to analyze the non-contact charging process and proposed a new design with a notch at the T-junction in aid of droplet splitting for more efficient charging. We experimentally characterized the induced charge in droplets in microfabricated devices. The experimental results agreed well with the simulation. Finally, we demonstrated highly effective droplet manipulation in a path selection unit appending to the droplet charging. We expect our work could enable precision manipulation of droplets for more complex liquid handling in microfluidics and promote electric-force based manipulation in 'lab-on-a-chip' systems.


In-vivo imaging of skin under stress: potential of high-frequency (20 MHz) static 2-D elastography.
The aim of this study was to evaluate the potential of high-frequency static two-dimensional (2-D) elastography for in vivo exploration of the mechanical behavior of skin. Our device was based on the combination of a 20 MHz sonographer and a patented extensiometer device able to apply calibrated uniaxial stretching of the skin. We used a new algorithm to compute elastograms that improve elastographic signal-to-noise ratio (SNRe) without sacrificing resolution. Mechanical behavior was described according to the axial strain and lateral displacements induced in the tissue. The efficacy of the strain anpolyvinyl alcohol first evaluated in polyvinyl alcohol (PVA)-cryogel phantoms. Several in vivo experiments then were conducted, mainly with the multistretching averaging method, and demonstrated the potential of this technique in the evaluation of mechanical behavior of the dermis and the hypodermis under stress.


Collagen fibre implant for tendon and ligament biological augmentation. In vivo study in an ovine model.
Although most in vitro studies indicate that collagen is a suitable biomaterial for tendon and ligament tissue engineering, in vivo studies of implanted collagen for regeneration of these tissues are still lacking. The objectives of this study were the following: (1) to investigate the regeneration of the central third of the ovine patellar tendon using implants made of an open array of collagen fibres (reconstituted, extruded bovine collagen); and (2) to compare two collagen crosslinking chemistries: carbodiimide and carbodiimide associated with ethyleneglycoldiglycidylether. Forty-eight Welsh Mountain sheep were operated on their right hind leg. The central third of patellar tendon was removed and substituted with carbodiimide (n = 16) and carbodiimide-ethyleneglycoldiglycidylether-crosslinked implants (n = 16). In the control group the defect was left empty (n = 16). The central third of contralateral unoperated tendons was used as positive controls. Half of the sheep in each group were killed at 3- and 6-month time points. After proper dissection, tendon sub-units (medial, central and lateral) were tested to failure (n = 6 for each group), whilst 2 non-dissected samples were used for histology. Both the implants had significantly lower stress to failure and modulus with respect to native tendon at both 3- and at 6-month time points. The implants did not statistically differ in stress to failure, whilst carbodiimide-crosslinked implants had significantly higher modulus than carbodiimide-ethyleneglycoldiglycidylether-crosslinked implants both at 3 and at 6 months. Histology showed carbodiimide-crosslinked implants to have a better integration with the native tendon than carbodiimide-ethyleneglycoldiglycidylether-crosslinked implants. Carbodiimide-crosslinked implants appeared partially resorbed and showed increased tissue ingrowth with respect to carbodiimide-ethyleneglycoldiglycidylether-crosslinked implants. To deliver collagen implants as an open array of fibres allows optimal tendon-implant integration and good ingrowth of regenerated tissue. In the present study the resorption rate of both the examined implants was too low due to the high level of crosslinking. This led to only minor substitution of the implant with regenerated tissue, which in turn produced a low-strength implanted region. Further studies are needed to find the right balance between strength and resorption rate of collagen fibres.


Is a Femoro-Acetabular Impingement Type Cam Predictable after Slipped Capital Femoral Epiphysis?
(1) Background: Previous studies have proven a high incidence of a femoro-acetabular impingement (FAI) type cam in patients sustaining a slipped capital femoral epiphysis (SCFE). Thus, the current study analyzed, if a cam deformity is predictable after SCFE treatment; (2) Methods: 113 cases of SCFE were treated between 1 January 2005 and 31 December 2017. The radiological assessment included the slip angle after surgery (referenced to the femoral neck (epiphyseal tilt) and shaft axis as Southwick angle) and the last available lateral center edge angle (LCEA), the acetabular- and alpha angle. A correlation was performed between these parameters and the last alpha angle to predict a FAI type cam; (3) Results: After a mean follow-up of 4.3 years (±1.9; 2.0-11.2), 48.5% of the patients showed a FAI type cam and 43.2% a dysplasia on the affected side. The correlation between the epiphyseal tilt and alpha angle was statically significant (p = 0.017) with a medium effect size of 0.28; (4) Conclusions: The postoperative posterior epiphyseal tilt was predictive factor to determine the alpha angle. However, the cut-off value of the slip angle was 16.8° for a later occurrence of a FAI type cam indicating a small range of acceptable deviations from the anatomical position for SCFE reconstruction.


High resolution photoemission and x-ray absorption spectroscopy of a lepidocrocite-like TiO2 nanosheet on Pt(110) (1 × 2).
The electronic structure of TiO(2) nanosheets on the Pt(110)-(1 × 2) surface has been investigated by using high resolution photoemission spectroscopy and x-ray absorption spectroscopy (XAS). The Ti 2p XAS spectra of the deposited TiO(2) films have been theoretically evaluated and, from the comparison with the experimental data, the assignment to a lepidocrocite-like structure is confirmed. Coexistence of TiO(2) islands with PtO(2) stripes for incomplete nanosheets is confirmed by high resolution photoemission data. The location of the valence and conduction band edges of the nanosheet has been experimentally determined allowing us to describe in details subtle electronic effects due to the interface with the substrate. The locations of the valence band maximum and the leading peak in the O 1s XAS spectrum indicate a band gap similar to anatase but with the Fermi level closer to mid-gap than found for bulk, n-type TiO(2).


Degradation of herpes simplex virions by human polymorphonuclear leukocytes and monocytes.
The degradation of herpes simplex virus particles after uptake by phagocytes was studied, but, since lysis of the phagocyte also resulted in damage to the viral envelope, measurement of viral infectivity as a criterion of viral degradation after phagocytosis was not possible. Therefore we focused on later events in viral destruction, namely the degradation of macromolecules. We have demonstrated that polymorphonuclear leukocytes (PMN) and monocytes (MN) can rapidly degrade the membrane proteins of the phagocytosed herpes-virus virions. PMN and MN from a patient with chronic granulomatous disease showed a similar rate of degradation compared to PMN and MN from healthy donors, which excludes an important role for toxic oxygen species in viral protein degradation. Experiments using toxic oxygen species-generating systems supported this observation. In contrast to PMN, MN are also effective in the digestion of viral DNA. We conclude that PMN and MN are able to neutralize large amounts of phagocytosed HSV, so their role in antiviral defence has again been demonstrated.


[Let us not resuscitate the dead].
Cardiopulmonary resuscitation (CPR) was developed as a preventive measure for sudden and unexpected death. This excellent technique has been misguided. It has been turned into an unmerited and cruel ritual in the horizon of all who are to die. CPR is particularly ineffective in the elderly. Of 503 patients age 69 or over subjected to CPR in 5 medical centers in Boston only 19 (3.8%) were alive at discharge. In 68.8% or them death occurred in less than 3 days. Only 2 of 244 patients who arrived at the hospital with a cardiorespiratory arrest survived; the 209 who arrived without vital signs died. One of 37 patients with an oxygen tension less than 63 torr left the hospital alive but connected to a respirator. All patients (204) with one or more of the following laboratory findings died: Hematocrit less than 35%, creatinine more than 1.5 mg/dl or a BUN more than 65 mg/ml, serum albumin less than 2.7 gr/dl. The rights of patients over their life and their history are violated when CPR is applied without their consent. In a recent study 63% of 104 hospitalized patients and 73% of 922 Puerto Rican doctors and nurses preferred not be subjected to CPR unless there was an opportunity for a complete functional recovery. Six percent would not have CPR under any circumstance. Health professionals should not continue avoiding their moral responsibilities shielded by institutional policies or hiding behind a simplistic religiosity. It is immoral to prolong the agony of dying patients. Let us cease the useless resuscitation of the dead.


Gestational Exposure to Bisphenol A and Bisphenol S Leads to Fetal Skeletal Muscle Hypertrophy Independent of Sex.
Gestational exposure to bisphenol A (BPA) can lead to offspring insulin resistance. However, despite the role that the skeletal muscle plays in glucose homeostasis, it remains unknown whether gestational exposure to BPA, or its analog bisphenol S (BPS), impairs skeletal muscle development. We hypothesized that gestational exposure to BPA or BPS will impair fetal muscle development and lead to muscle-specific insulin resistance. To test this, pregnant sheep (n = 7-8/group) were exposed to BPA or BPS from gestational day (GD) 30 to 100. At GD120, fetal skeletal muscle was harvested to evaluate fiber size, fiber type, and gene and protein expression related to myogenesis, fiber size, fiber type, and inflammation. Fetal primary myoblasts were isolated to evaluate proliferation and differentiation. In fetal skeletal muscle, myofibers were larger in BPA and BPS groups in both females and males. BPA females had higher MYH1 (reflective of type-IIX fast glycolytic fibers), whereas BPS females had higher MYH2 and MYH7, and higher myogenic regulatory factors (Myf5, MyoG, MyoD, and MRF4) mRNA expression. No differences were observed in males. Myoblast proliferation was not altered in gestationally BPA- or BPS-exposed myoblasts, but upon differentiation, area and diameter of myotubes were larger independent of sex. Females had larger myofibers and myotubes than males in all treatment groups. In conclusion, gestational exposure to BPA or BPS does not result in insulin resistance in fetal myoblasts but leads to fetal fiber hypertrophy in skeletal muscle independent of sex and alters fiber type distribution in a sex-specific manner.


Decreased functional connectivity and structural deficit in alertness network with right-sided temporal lobe epilepsy.
Patients with temporal lobe epilepsy (TLE) often suffer from alertness alterations. However, specific regions connected with alertness remain controversial, and whether these regions have structural impairment is also elusive. This study aimed to investigate the characteristics and neural mechanisms underlying the functions and structures of alertness network in patients with right-sided temporal lobe epilepsy (rTLE) by performing the attentional network test (ANT), resting-state functional magnetic resonance imaging (R-SfMRI), and diffusion tensor imaging (DTI).A total of 47 patients with rTLE and 34 healthy controls underwent ANT, R-SfMRI, and DTI scan. The seed-based functional connectivity (FC) method and deterministic tractography were used to analyze the data.Patients with rTLE had longer reaction times in the no-cue and double-cue conditions. However, no differences were noted in the alertness effect between the 2 groups. The patient group had lower FC compared with the control group in the right inferior parietal lobe (IPL), amygdala, and insula. Structural deficits were found in the right parahippocampal gyrus, superior temporal pole, insula, and amygdala in the patient group compared with the control group. Also significantly negative correlations were observed between abnormal fractional anisotropy (between the right insula and the superior temporal pole) and illness duration in the patients with rTLE.The findings of this study suggested abnormal intrinsic and phasic alertness, decreased FC, and structural deficits within the alerting network in the rTLE. This study provided new insights into the mechanisms of alertness alterations in rTLE.


Long-term complications associated with prosthetic repair of incisional hernias.
To determine whether the type of prosthetic material and technique of placement influenced long-term complications after repair of incisional hernias. Retrospective cohort analytic study. University-affiliated hospital. Two hundred patients undergoing open repair of abdominal incisional hernias with prosthetic material between 1985 and 1994. Four types of prosthetic material were used and placed either as an onlay, underlay, sandwich, or finger interdigitation technique. The materials were monofilamented polypropylene mesh (Marlex, Davol Inc, Cranston, RI), double-filamented mesh (Prolene, Ethicon Inc, Somerville, NJ), expanded polytetrafluroethylene patch (Gore-Tex, WL Gore & Associates, Phoenix, Ariz) or multifilamented polyester mesh (Mersilene, Ethicon Inc). The incidence of recurrence and complications such as enterocutaneous fistula, bowel obstruction, and infection with each type of material and technique of repair were compared with univariate and multivariate analysis. On univariate analysis, multifilamented polyester mesh had a significantly higher mean number of complications per patient (4.7 vs 1.4-2.3; P<.002), a higher incidence of fistula formation (16% vs 0%-2%; P<.001), a greater number of infections (16% vs 0%-6%; P<.05), and more recurrent hernias (34% vs 10%-14%; P<.05) than the other materials used. The additional mean length of stay to treat complications was also significantly longer (30 vs 3-7 days; P<.001) when polyester mesh was used. The deleterious effect of polyester mesh on long-term complications was confirmed on multiple logistic regression (P=.002). The technique of placement had no influence on outcome. Polyester mesh should no longer be used for incisional hernia repair.


Long-term improvement in unsafe injection practices following community intervention.
A study in 1994 identified frequent unsafe injections as the cause of widespread hepatitis C virus infection in Hafizabad, Pakistan. A simple low cost community education program was assessed to see if it improved injection safety. A local health organization developed educational materials on hepatitis C including advice on how to avoid unnecessary injections and, when injections were necessary, to use a new syringe and needle. Beginning in 1995, this advice was communicated through multiple channels including health education meetings, announcements in mosques, and via pamphlets. In 1998 study workers revisited controls from the 1994 case-control study (along with three of their neighbors of a similar age) to collect information on injection practices in the previous 12 months. Thirty-three percent of the study's participants in 1998 received >or=5 injections in the preceding 12 months compared to 40% of the hepatitis C virus negative controls reported in the year prior to the 1994 study (p=0.85). In 1998 52 persons (34%) brought their own syringe for their most recent injection, a practice that was unreported in 1994. Overall, in 1998 59% of patients received their most recent injection with a new syringe and needle compared to 24% in 1994 (p=0.003). Following this low cost health communication effort, community members took steps to protect themselves from unsafe injections.


Induction of secretogranin II mRNA by protein synthesis inhibitors in GH4C1 cells.
The effect of cycloheximide (CHX) on the expression of secretogranin II (SgII), a member of the granin family of secretory proteins, was investigated in rat pituitary GH4C1 (GH) cells. The administration of CHX resulted in a dose- and time-dependent increase in SgII mRNA expression, the greatest effect (3.8-fold above control) being achieved with 1 microgram/ml CHX, which resulted in > 90% inhibition of protein synthesis. Emetine (1 microgram/ml), pactamycin (0.6 microgram/ml), anisomycin (2.5 micrograms/ml), and puromycin (100 micrograms/ml), protein synthesis inhibitors structurally and mechanistically unrelated to CHX, also increased the level of SgII mRNA expression. Treatment with forskolin (10 microM) alone had no effect on SgII mRNA levels but potentiated the effect of CHX. Neither 100 nM phorbol 12-myristate 13-acetate nor 45 mM KCl affected SgII mRNA levels in the absence or presence of 1 microgram/ml CHX. The effect of CHX was blocked by the transcription inhibitors actinomycin D (5 micrograms/ml) and 5,6-dichlorobenzimidazole riboside (20 micrograms/ml) but not by the coadministration of the polysome destabilizer pactamycin (0.6 microgram/ml), suggesting that the effect of CHX was transcriptional. These studies show that the expression of SgII mRNA is induced by protein synthesis inhibitors in GH cells, suggesting the presence of a labile repressor, which not only controls the basal expression of the SgII gene but also completely inhibits the stimulatory effect of forskolin.


Clonal Spread and Intra- and Inter-Species Plasmid Dissemination Associated With Klebsiella pneumoniae Carbapenemase-Producing Enterobacterales During a Hospital Outbreak in Barcelona, Spain.
Objectives: The study aimed to characterize the clonal spread of resistant bacteria and dissemination of resistance plasmids among carbapenem-resistant Enterobacterales at a tertiary hospital in Catalonia, Spain. Methods: Isolates were recovered from surveillance rectal swabs and diagnostic samples. Species identification was by matrix-assisted laser desorption ionization-time time of flight mass spectrometry (MALDI-TOF MS). Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Antimicrobial susceptibility was assessed by gradient-diffusion and carriage of bla genes was detected by PCR. Plasmid typing, conjugation assays, S1-PFGE studies and long-read sequencing were used to characterize resistance plasmids. Results: From July 2018 to February 2019, 125 Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacterales were recovered from 101 inpatients from surveillance (74.4%) or clinical samples (25.6%), in a tertiary hospital in Barcelona. Clonality studies identified a major clone of Klebsiella pneumoniae belonging to sequence type ST15 and additional isolates of K. pneumoniae, Escherichia coli and Enterobacter sp. from different STs. All isolates but one carried the bla KPC-2 allelic variant. The bla KPC-2 gene was located in an IncFIIk plasmid of circa 106 Kb in a non-classical Tn4401 element designated NTEKPC-pMC-2-1. Whole-genome sequencing revealed different rearrangements of the 106 Kb plasmid while the NTEKPC-pMC-2-1 module was highly conserved. Conclusion: We report a hospital outbreak caused by the clonal dissemination of KPC-producing ST15 K. pneumoniae but also the intra- and inter-species transmission of the bla KPC-2 gene associated with plasmid conjugation and/or transposon dissemination. To our knowledge, this is the first report of an outbreak caused by KPC-producing Enterobacterales isolated from human patients in Catalonia and highlights the relevance of surveillance studies in the early detection and control of antibiotic resistant high-risk clones.


Capsaicin in the Treatment of Refractory Neuropathic Pain after Mastectomy Surgery: A Case Report.
A 62-year-old female patient with a history of mastectomy surgery and sentinel lymphadenectomy in the context of breast cancer therapy was referred to our clinic for the treatment of refractory neuropathic pain. She reported a complex set of symptoms including burning and electrical-like sensations as well as profound hyperesthesia, hyperalgesia, and allodynia. The symptoms persisted chronically over months with a strong intensity and did not sufficiently respond to oral pain medication and co-analgetics, that is, tapentadol and pregabalin. As the patient could hardly move her right upper arm due to the pain, the quality of life was greatly reduced. In addition, the patient reported pain-related anxiety and depression. Therefore, a therapy with capsaicin 8% patch was initiated. Treatment with capsaicin 8% led to pain relief without tolerance development and improved flexibility in the affected body area. Despite significant pain relief, previous oral pain medications (tapentadol, pregabalin) as well as the anti-depressant amitriptyline were maintained to fully resolve pain symptoms, anxiety, and depression. In conclusion, capsaicin 8% may represent an effective therapeutic alternative for patients suffering from refractory neuropathic pain.


Imaging in low-grade glioma: a guide for neurologists.
The management of low-grade glioma (LGG) is shifting as evidence has emergedthat refutes the previously commonplace imaging-based 'watch and wait' approach, in favour of early aggressive surgical resection. This coupled with the recent 2016 update to the World Health Organisation Classification of Tumours of the Central Nervous System is changing LGG imaging and management. Recently in Practical Neurology the contemporary management of low-grade glioma and the changes to this grading system were discussed in detail. 1 In this complementary article, we discuss the role of imaging in the diagnosis, surgical planning and post-treatment follow-up of LGG. We describe the principles of imaging these tumours and use several cases to highlight some difficult scenarios.


Selective localization of mesenteric lymphoblasts in mucosal tissues: effects of altering the number of donor lymphoblasts.
We have studied the effect of altering the numbers of lymphoblasts from the mesenteric lymph node (MLN) transferred into syngeneic female CBA/J mice on their distribution and abundance 24 hr later. The frequency of [3H]-thymidine-labelled donor MLN cells in the recipient small intestine and lungs was directly related to the numbers transferred. Of the donor MLN lymphoblasts in the small intestine, 62.5% +/- 0.9% were seen in the basal lamina propria, 32.5% +/- 0.9% in the villus lamina propria and 5% +/- 0.6% in the epithelium. Of the MLN lymphoblasts localizing in the lungs, 90% +/- 2.3% were in the parenchyma while 6.7% +/- 1.8% and 3.3% +/- 1.0% appeared in the bronchus-associated lymphoid tissue (BALT) and the bronchial epithelium, respectively. Although few peripheral lymph node (PLN)- derived lymphoblasts localized in the small intestine, the numbers of PLN lymphoblasts in the lungs were similar to those observed after transfer of comparable doses of MLN lymphoblasts. However, PLN Lymphoblasts were found only in the pulmonary parenchyma and did not appear in either BALT or bronchial epithelium. These data suggest that the number of MLN, but likely not PLN, lymphoblasts in the circulation, directly influences the numbers of lymphoblasts which localize in intestinal mucosa, BAlt and bronchial epithelium. Even at the highest doses of MLN lymphoblasts transferred we could not saturate the capacity of these tissues to accommodate MLN lymphoblasts nor was their intra-intestinal distribution altered.


[The effect of proteolysis of myosin heads on conformational changes induced by them in F-actin].
The effect of limited proteolysis of myosin subfragment I (S1) on conformational changes in F-actin during the formation of a rigor F-actin-S1 complex was studied, using polarized microfluorimetry. Upon the decoration of thin filaments made up of rhodaminyl-phalloin modified F-actin with subfragment I, the anisotropy of fluorescence increased. Limited proteolysis of S1 at the junctions of 27 kD-70 kD and 27 kD-50 kD-20 kD fragments as well as the destruction of the 50 kD fragment by methanol had no effect on the nature of these changes. It was assumed that during the formation of the actomyosin complex the conformational changes in F-actin were induced by the 20 kD fragment. The 27 kD and 50 kD fragments seemed to exert a weak influence on action conformation within the complex.


In vivo lymphocyte tropism of feline immunodeficiency virus.
Feline immunodeficiency virus (FIV) infection in the cat is similar to human immunodeficiency virus type 1 infection in causing a selective reduction in CD4+ cell numbers, leading to inversion of the CD4+/CD8+ ratio. To determine whether FIV, similar to human immunodeficiency virus type 1, has a tropism for CD4+ cells, we examined the in vitro and in vivo susceptibilities of feline lymphocyte subpopulations to FIV infection. Infection of interleukin-2-dependent CD4+ or CD8+ lymphocyte cultures with the NCSU1 isolate of FIV (FIV-NCSU1) resulted in syncytium formation, cell death, and Mg(2+)-dependent reverse transcriptase (RT) activity in both cases. Monoclonal antibodies to feline lymphocyte subsets were used to sort peripheral blood mononuclear cells from FIV-infected cats into highly (> 95%) purified CD4+ cell, CD8+ cell, immunoglobulin-positive (Ig+) cell, and monocyte subpopulations. The mononuclear cell subpopulations were analyzed for FIV provirus by polymerase chain reaction and Southern blot analysis and for virus expression by RT activity. All 16 cats infected with FIV-NCSU1 demonstrated FIV provirus in CD4+ cell-, CD8+ cell-, and Ig+ cell-enriched lymphocyte populations. Southern blot detection of amplified gag gene sequences and limiting-cell-dilution polymerase chain reaction analysis indicated that Ig+ cells carried a higher FIV provirus burden in chronically (> or = 1-year) infected cats than either CD4+ or CD8+ cells. In contrast, CD4+ cells carried the greatest provirus burden in acutely (2- to 4-week) infected cats. FIV provirus was detected in monocytes from only 1 of 10 cats with asymptomatic infection. Addition of culture supernatants from enriched CD4+, CD8+, and Ig+ cells from FIV-infected cats to an FIV-susceptible CD4+ lymphocyte culture resulted in syncytium formation, cell death, and RT activity. Infection of Ig+ cells is not unique to FIV-NCSU1, as lymphocyte subpopulations from other cats with natural infections and cats infected with the Petaluma or Mount Airy isolate of FIV demonstrated a similar distribution of FIV provirus and RT activity. These data suggest that FIV possesses a broad tropism for peripheral blood mononuclear cells and that an Ig+ cell may serve as a major reservoir for the virus in chronically infected cats.


Intracorporeal versus extracorporeal anastomosis after laparoscopic left colectomy for splenic flexure cancer: results from a multi-institutional audit on 181 consecutive patients.
Although intracorporeal anastomosis has been demonstrated to be safe and effective after right colectomy, limited data are available about its efficacy after left colectomy for colon cancer located in splenic flexure. A multi-institutional audit was designed, including 92 patients who underwent laparoscopic left colectomy with intracorporeal anastomosis (IA) compared with 89 matched patients who underwent a laparoscopic left colectomy with extracorporeal anastomosis (EA). There was no significant difference in terms of age, sex, BMI, and ASA score between the two groups. Post-surgical history and stage of disease according to AJCC/UICC TNM were also similar. IA and EA groups demonstrated similar oncologic radicality in terms of the number of lymph nodes harvested (18.5 ± 9 vs. 17.5 ± 8.4; p = 0.48). Recovery after surgery was also better in patients who underwent IA, as confirmed by the shorter time to flatus in the IA group (2.6 ± 1.1 days vs. 3.4 ± 1.2 days; p < 0.001) and higher post-operative pain expressed in the mean VAS Scale in the EA group (1.7 ± 2.1 vs. 3.5 ± 1.6; p < 0.001). Laparoscopic left colectomy with intracorporeal anastomosis was associated with a lower rate of post-operative complications (OR 6.7, 95% CI 2.2-20; p = 0.001). However, when stratifying according to Clavien classification, the difference was consistently confirmed for less severe (class I and II) complications (OR 7.6, 95% CI 2.5-23, p = 0.001) but not for class III, IV, and V complications (OR 1.8, 95% CI 0.1-16.9; p = 0.59). Our results were consistent to hypothesize that a complete laparoscopic approach could be considered a safe method to perform laparoscopic left colectomy with the advantage of a guaranteed faster recovery after surgery. Further randomized clinical trials are needed to obtain a more definitive conclusion.


spongEffects: ceRNA modules offer patient-specific insights into the miRNA regulatory landscape.
Cancer is one of the leading causes of death worldwide. Despite significant improvements in prevention and treatment, mortality remains high for many cancer types. Hence, innovative methods that use molecular data to stratify patients and identify biomarkers are needed. Promising biomarkers can also be inferred from competing endogenous RNA (ceRNA) networks that capture the gene-miRNA gene regulatory landscape. Thus far, the role of these biomarkers could only be studied globally but not in a sample-specific manner. To mitigate this, we introduce spongEffects, a novel method that infers subnetworks (or modules) from ceRNA networks and calculates patient- or sample-specific scores related to their regulatory activity. We show how spongEffects can be used for downstream interpretation and machine learning tasks such as tumor classification and for identifying subtype-specific regulatory interactions. In a concrete example of breast cancer subtype classification, we prioritize modules impacting the biology of the different subtypes. In summary, spongEffects prioritizes ceRNA modules as biomarkers and offers insights into the miRNA regulatory landscape. Notably, these module scores can be inferred from gene expression data alone and can thus be applied to cohorts where miRNA expression information is lacking. https://bioconductor.org/packages/devel/bioc/html/SPONGE.html.


Presentations to an acute medical unit due to headache: a review of 306 consecutive cases.
The United Kingdom National Health Service has recently prioritised the need for ambulatory care pathways for acute headache. The present study sought to better characterise patients referred to an Acute Medical Unit so as to inform pathway development. In 2011, York Hospital received 306 referrals due to acute headache, representing 3% of acute medical admissions. Investigations included CT scan (38%), lumbar puncture (38%), and MRI (18%); there were no specialised investigations in 26%, and 18% of patients were discharged on the day of presentation. Subarachnoid haemorrhage occurred in only 4 patients (1%), meningitis in 10 (3%), and intracranial tumour in 5 (2%). The findings indicate that a significant proportion of patients with acute headache could be managed by ambulatory care.


Interspecific differences in egg production affect egg trace element concentrations after a coal fly ash spill.
In oviparous vertebrates, trace elements transfer from mother to offspring during egg production. For animals that produce eggs slowly, like turtles, the trace element concentration of each egg reflects an integration of dietary and stored accumulation over the duration of vitellogenesis. Because turtles also produce eggs synchronously, all eggs within a clutch should exhibit uniform trace element concentrations. In contrast, for animals that produce eggs in sequence and primarily from current dietary resources, like many birds, the trace element concentrations of eggs should be less uniform within a clutch, and likely reflect short-term changes in dietary exposure. We tested the hypothesis that stinkpot turtle (Sternotherus odoratus) clutches exhibit lower variability and higher repeatability in barium, selenium, strontium, and thallium concentrations than those of tree swallows (Tachycineta bicolor) from a site impacted by a recent coal ash spill. All four trace elements exhibited significantly lower variability and significantly higher repeatability in stinkpot clutches than in swallow clutches. Mean trace element concentrations of stinkpot eggs were also significantly higher than those of swallow eggs although both species feed primarily on aquatic invertebrates. Variability in swallow egg trace element concentrations was partially due to significant laying order effects. Our results support the hypothesis that interspecific variation in the source of resources and in the synchronicity and rate of egg production can lead to interspecific differences in the variability of egg trace element concentrations.


Ablation of atrial fibrillation: patient selection, techniques, and the results.
Atrial fibrillation (AF) is a common arrhythmia, affecting an estimated 2 million people in the United States and its prevalence increases with age, reaching 10% in those > or = 80 years. AF confers a four- to fivefold increased risk of stroke compared to the general population and has been associated with a doubling of all-cause mortality. During the past decade, limited success rates of drug treatment stimulated an exploration of interventional treatment options for AF. As our knowledge on initiating triggers and perpetuating substrate of AF expanded, different potentially curative catheter ablation techniques have been developed. In this article we review the current patient selection criteria, methods, and the results of the catheter ablation of atrial fibrillation.


Changes in sterol and phospholipid fatty acid composition in Refsum's disease fibroblasts grown in the presence of phytol.
Fibroblasts derived from an individual with Refsum's disease (GM 3896) and a normal control (GM 1717) were grown in the presence of 0, 0.1, 0.2, and 0.3 mM phytol. Cultures were analyzed for total sterol content, and the fatty acid composition of the extractable phospholipids. The fatty acid composition of the phospholipids are similar for control and Refsum's disease fibroblasts when grown on media lacking phytol. However, the addition of phytol to the growth medium produces differences in fatty acid composition between the phospholipids extracted from control and Refsum's disease cells. With regard to sterol composition, data are presented which suggest that an altered sterol is induced in Refsum's disease fibroblasts by the presence of phytol in the growth medium. The possible relationship of these findings to the mechanism of Refsum's disease is discussed.


Spectroscopic Differentiation and Microscopic Imaging of Red Organic Pigments Using Optical Pump-Probe Contrast.
Analysis of red organic pigments in artworks (and in forensics applications) is challenging, because conventional nondestructive mapping techniques provide little contrast, and most chemical analyses with high specificity require sample removal. Here we demonstrate a new optical approach, pump-probe microscopy, for the analysis of red organic pigments. We investigate Carmine naccarat, Lac dye, purpurin, alizarin, madder lake, and eosin Y and show that their intrinsic photophysical properties produce distinctive pump-probe spectra. We utilize this contrast for high-resolution, three-dimensional imaging without the need for physical sample removal. Lastly, we highlight the potential of pump-probe microscopy as an analytical tool for forensics of other types of organic colorants by investigating a series of automotive paints.


G protein γ subunit qPE9-1 is involved in rice adaptation under elevated CO2 concentration by regulating leaf photosynthesis.
G protein γ subunit qPE9-1 plays multiple roles in rice growth and development. However, the role of qPE9-1 in rice exposed to elevated carbon dioxide concentration (eCO2) is unknown. Here, we investigated its role in the regulation of rice growth under eCO2 conditions using qPE9-1 overexpression (OE) lines, RNAi lines and corresponding WT rice. Compared to atmospheric carbon dioxide concentration (aCO2), relative expression of qPE9-1 in rice leaf was approximately tenfold higher under eCO2. Under eCO2, the growth of WT and qPE9-1-overexpressing rice was significantly higher than under aCO2. Moreover, there was no significant effect of eCO2 on the growth of qPE9-1 RNAi lines. Furthermore, WT and qPE9-1-overexpressing rice showed higher net photosynthetic rate and carbohydrate content under eCO2 than under aCO2. Moreover, the relative expression of some photosynthesis related genes in WT, but not in RNAi3 line, showed significant difference under eCO2 in RNA-seq analysis. Compared to WT and RNAi lines, the rbcL gene expression and Rubisco content of rice leaves in qPE9-1-overexpressors were higher under eCO2. Overall, these results suggest that qPE9-1 is involved in rice adaptation under elevated CO2 concentration by regulating leaf photosynthesis via moderating rice photosynthetic light reaction and Rubisco content.