Mice intranasally immunized with a recombinant 16-kilodalton antigen from roundworm Ascaris parasites are protected against larval migration of Ascaris suum.
Protective immunity to the pig roundworm, Ascaris suum, has been demonstrated by immunization of pigs with antigens derived from the parasite's larval stages. We identified a protective antigen commonly expressed in the human and pig Ascaris infections as a 16-kDa protein (As16), which has no similarity at the amino acid level to mammalian proteins but has some similarity to those of the filarial parasites and Caenorhabditis elegans gene product. Localization analysis revealed that the native As16 was highly expressed in the adult worm intestine, hypodermis, and cuticles. In addition, As16 was detected in the parasite excretory and secretory products. Mice intranasally vaccinated with Escherichia coli-expressed recombinant As16 (rAs16), coupled with cholera toxin B subunit, generated a significant increase in the level of rAs16-specific immunoglobulin G (IgG) and IgE in serum. Mucosal IgA levels were also increased. The recombinant protein evoked a mixed (both Th1 and Th2) type of immune response characterized by elevated levels of gamma interferon and interleukin-10 in the culture supernatants of activated spleen cells. An increased level of IgG1 and IgG2a in serum was also observed. The vaccinated mice showed a reduction by 58% in the recovery of challenged larvae compared to a nonvaccinated control. These results suggest the possibility of developing a mucosal vaccine for human and pig ascariasis.


Polymerase chain reaction-based identification and genotyping of Anopheles mosquitoes with a 96-pin bacterial replicator.
A simple method for rapid identification of large numbers of Anopheles mosquitoes was developed based on polymerase chain reaction (PCR) amplification of the rDNA intergenic spacer and internal transcribed spacer 2. By means of previously described primers for the Anopheles gambiae and An. quadrimaculatus species complexes, rDNA was amplified simultaneously from 96 whole mosquitoes or parts. No homogenization or individual DNA preparation was necessary, and transfer of 96 samples to PCR reactions was performed simultaneously with a bacterial replicator. Control reactions indicate that the level of cross-contamination is negligible, and false-negative findings are rare. The method was tested on larvae, pupae, adult heads, whole adult males and females, and single tarsi. All parts except tarsi provided satisfactory template. Fresh, ethanol-preserved, dried, and frozen adults were also tested with similar results. The method was also tested for amplification of a single-copy gene, white. Results were generally positive, although some false-negative findings were observed. This method allows rapid analysis of large numbers of mosquitoes without robotic equipment and should enable rapid and extensive PCR analysis of field-collected samples and laboratory specimens.


T cell-macrophage interactions and granuloma formation in vasculitis.
Granuloma formation, bringing into close proximity highly activated macrophages and T cells, is a typical event in inflammatory blood vessel diseases, and is noted in the name of several of the vasculitides. It is not known whether specific properties of the microenvironment in the blood vessel wall or the immediate surroundings of blood vessels contribute to granuloma formation and, in some cases, generation of multinucleated giant cells. Granulomas provide a specialized niche to optimize macrophage-T cell interactions, strongly activating both cell types. This is mirrored by the intensity of the systemic inflammation encountered in patients with vasculitis, often presenting with malaise, weight loss, fever, and strongly upregulated acute phase responses. As a sophisticated and highly organized structure, granulomas can serve as an ideal site to induce differentiation and maturation of T cells. The granulomas possibly seed aberrant Th1 and Th17 cells into the circulation, which are known to be the main pathogenic cells in vasculitis. Through the induction of memory T cells, aberrant innate immune responses can imprint the host immune system for decades to come and promote chronicity of the disease process. Improved understanding of T cell-macrophage interactions will redefine pathogenic models in the vasculitides and provide new avenues for immunomodulatory therapy.


Efficacy of the decolonization of methicillin-resistant Staphylococcus aureus carriers in clinical practice.
Nasal and extra nasal carriage of methicillin-resistant S. aureus (MRSA) is a pre-existing condition that often leads to invasive MRSA infection, as MRSA colonization is associated with a high risk of acquiring MRSA infection during hospital stays. Decolonization may reduce the risk of meticillin-resistant Staphylococcus aureus (MRSA) infection in individual carriers and prevent transmission to other patients. A retrospective cohort study was conducted to evaluate the effectiveness of two decolonization protocols for newly diagnosed MRSA carriage in hospitalized patients and to assess the impact of decolonization on the rate of MRSA infection. The study population consisted of all patients diagnosed as MRSA-positive between January 2006 and June 2010. Patients diagnosed as carriers were designated as requiring contact precautions by the hospital infection control team. The standing order protocol of the hospital pertaining to decolonization procedures was then applied, and all newly diagnosed patients were administered one of the two decolonization treatments outlined in the hospital protocol, with the exception of MRSA respiratory carriers (MRSA obtained from sputum or other lower respiratory tract samples). The two decolonization treatments consisted of the application of intranasal mupirocin 2 % and washing with chlorhexidine soap (40 mg/mL) (mupi/CHX) or application of intranasal povidone-iodine and washing with povidone-iodine soap (PVPI), with each treatment lasting for 5 days. Success was determined by at least three successive nose swabs and throat and other screened site swabs that tested negative for MRSA before patient discharge. A total of 1150 patients admitted to the hospital were found to be infected or colonized with MRSA. Of the 1150 patients, 268 were prescribed decolonization treatment. 104 out of 268 patients (39 %) were successfully decolonized. There was no significant success after two decolonization failures. MRSA infection rate among the successes and failures were 0.0 and 4.3 %, respectively [P = 0.04]. Our results fit well with the prescription of decolonization based on local strategy protocols but reflect a low rate of successful treatment. Although the success rate of decolonization was not high in our study, the effectiveness of decolonization on the infection rate, justifies the continuation of this strategy, even if a marginal cost is incurred.


Uncemented total hip arthroplasty for primary osteoarthritis in young patients: a mid-to long-term follow-up study from the Finnish Arthroplasty Register.
The survival of total hip arthroplasties (THAs) has been considered to be poor in young patients. We evaluated the population-based survival of uncemented THA for primary osteoarthritis (OA) in patients under 55 years of age and the factors affecting survival. The Finnish Arthroplasty Register was established in 1980. Between that year and 2003, 92,083 primary THAs were entered in the register, 5,607 of which were performed for primary OA in patients under 55 years of age. Using records from these 5,607 THAs, we selected uncemented femoral and acetabular components that had been used in more than 100 operations during the study period. Survival of both components (cup/stem) and their combinations were analyzed separately with the Kaplan-Meier analysis and the Cox regression model. All uncemented stems studied showed a survival rate of over 90% at 10 years. The Biomet Bi-Metric stem had a 95% (95% CI 93- 97) survival rate even at 15 years. Overall survival of the extendedly porous-coated Lord Madréporique stem (p = 0.003) and the proximally porous-coated Anatomic Mesh stem (p = 0.0008) were poorer than that of the Biomet Bi-Metric stem. When endpoint was defined as stem revision for any reason, results were generally similar; there was no difference, however, between the survival rates of the Lord Madréporique stem and the Bi-Metric stem. Of the acetabular components, the Biomet Universal, the ABG Il and the Harris-Galante II cups showed < 90% survival rates at 10 years with aseptic loosening as endpoint; at 13 years the corresponding survival rates were 94% (95% CI 91-97) for the Biomet Universal and 95% (95% CI 91-98) for the Harris-Galante II cups with aseptic loosening as endpoint. The PCA Pegged porous-coated uncemented cup showed a poor 13-year survival rate of 68% (95% CI 59-78) with aseptic loosening as endpoint. However, when endpoint was defined as any revision (including exchange of liner), the 10-year survival rates of all brands of cup except Harris-Galante II declined to under 80%. Modern second-generation uncemented stems, with proximal circumferential porous- or HA-coating, seem to be a good choice for young patients with primary OA. Similarly, modern press-fit porous- and HA-coated cups appear to have good endurance against aseptic loosening in these young patients. However, liner revisions were common; thus, survival rates of uncemented cups were unsatisfactorily low. Polyethylene wear and unfavorable locking mechanisms between the metal shell and the polyethylene liner and their sequelae remain matters of concern in this young and active group of patients.


Host genetic factors and vaccine-induced immunity to HBV infection: haplotype analysis.
Hepatitis B virus (HBV) infection remains a significant health burden world-wide, although vaccines help decrease this problem. We previously identified associations of single nucleotide polymorphisms in several candidate genes with vaccine-induced peak antibody level (anti-HBs), which is predictive of long-term vaccine efficacy and protection against infection and persistent carriage; here we report on a haplotype-based analysis. A total of 688 SNPs from 117 genes were examined for a two, three and four sliding window haplotype analysis in a Gambian cohort. Analysis was performed on 197 unrelated individuals, 454 individuals from 174 families, and the combined sample (N = 651). Global and individual haplotype association tests were carried out (adjusted for covariates), employing peak anti-HBs level as outcome. Five genes (CD44, CD58, CDC42, IL19 and IL1R1) had at least one significant haplotype in the unrelated or family analysis as well as the combined analysis. Previous single locus results were confirmed for CD44 (combined global p = 9.1x10(-5) for rs353644-rs353630-rs7937602) and CD58 (combined global p = 0.008 for rs1414275-rs11588376-rs1016140). Haplotypes in CDC42, IL19 and IL1R1 also associated with peak anti-HBs level. We have identified strong haplotype effects on HBV vaccine-induced antibody level in five genes, three of which, CDC42, IL19 and IL1R1, did not show evidence of association in a single SNP analyses and corroborated the majority of these effects in two datasets. The haplotype analysis identified associations with HBV vaccine-induced immunity in several new genes.


Development and validation of a nomogram based on clinical factors and standard laboratory tests for prediction of clinically significant liver fibrosis in chronic hepatitis C virus infection.
Staging liver fibrosis in chronic viral hepatitis C (HCV) patients is essential for prompting surveillance and treatment. The aim of this study was to develop a nomogram, on the basis of simple clinical and laboratory variables, to predict three clinically significant stages of fibrosis (nil-mild, moderate, advanced/cirrhosis), using histology as reference, and to compare its performance with that of FibroTest, a widely used noninvasive fibrosis score. Nomograms are graphical representations of a mathematical formula, used as predictive tools. The study retrospectively recruited 406 HCV patients undergoing liver biopsy. Nomogram was developed in a training set of 252 patients and tested in a validation set of 154 patients. Histology was staged according to the Metavir system. Fibrosis stages were subgrouped as follows: advanced fibrosis/cirrhosis (F3/F4, 24%), nil-mild (F0/F1, 36%), and moderate (F2, 40%). Age at biopsy, aspartate aminotransferase, γ-glutamyl transpeptidase, albumin, platelet count, and prothrombin activity formed the basis for the so-called Fibro-Nomogram, which, in one graphical representation, estimates probability for different stages of fibrosis. Areas under the receiver-operating characteristic curves for advanced fibrosis/cirrhosis were similar for training (0.86) and validation sets (0.87). For nil-mild fibrosis, area under the receiver-operating characteristics were 0.81 and 0.79. Compared with FibroTest, Fibro-Nomogram performed slightly better at predicting severe fibrosis (F3/F4) with positive likelihood ratio (LR+) 5.07 (95% confidence interval 3.08-8.37) versus LR+ 3.82 (95% confidence interval 2.56-5.71) for FibroTest. For nil-mild fibrosis, the two tests showed limited but comparable performances. In HCV patients, Fibro-Nomogram, an inexpensive and readily available predictive tool, could enable clinicians to interpret patients' profile, concurrently stratifying patients into three clinically relevant probability categories with good overall performance.


P wave dispersion is increased in pulmonary stenosis.
The right atrium pressure load is increased in pulmonary stenosis (PS) that is a congenital anomaly and this changes the electrophysiological characteristics of the atria. However, there is not enough data on the issue of P wave dispersion (PWD) in PS. Forty-two patients diagnosed as having valvular PS with echocardiography and 33 completely healthy individuals as the control group were included in the study. P wave duration, p wave maximum (p max) and p minimum (p min) were calculated from resting electrocardiography (ECG) obtained at the rate of 50 mm/sec. P wave dispersion was derived by subtracting p min from p max. The mean pressure gradient (MPG) at the pulmonary valve, structure of the valve and diameters of the right and left atria were measured with echocardiography. The data from two groups were compared with the Mann-Whitney U test and correlation analysis was performed with the Pearson correlation technique. There wasn't any statistically significance in the comparison of age, left atrial diameter and p min between two groups. While the MPG at the pulmonary valve was 43.11 +/- 18.8 mmHg in PS patients, it was 8.4 +/- 4.5 mmHg in the control group. While p max was 107.1 +/- 11.5 in PS group, it was 98.2 +/- 5.1 in control group (p=0.01), PWD was 40.4 +/- 1.2 in PS group, and 27.2 +/- 9.3 in the control group (p=0.01). Moreover, while the diameter of the right atrium in PS group was greater than that of the control group, (38.7 +/- 3.9 vs 30.2 +/- 2.5, p=0.02). We detected a correlation between PWD and pressure gradient in regression analysis. P wave dispersion and p max are increased in PS. While PWD was correlated with the pressure gradient that is the degree of narrowing, it was not correlated with the diameters of the right and left atria.


Inhibition of class I HDACs abrogates the dominant effect of MLL-AF4 by activation of wild-type MLL.
The ALOX5 gene encodes 5-lipoxygenase (5-LO), a key enzyme of inflammatory reactions, which is transcriptionally activated by trichostatin A (TSA). Physiologically, 5-LO expression is induced by calcitriol and/or transforming growth factor-β. Regulation of 5-LO mRNA involves promoter activation and elongation control within the 3'-portion of the ALOX5 gene. Here we focused on the ALOX5 promoter region. Transcriptional initiation was associated with an increase in histone H3 lysine 4 trimethylation in a TSA-inducible manner. Therefore, we investigated the effects of the MLL (mixed lineage leukemia) protein and its derivatives, MLL-AF4 and AF4-MLL, respectively. MLL-AF4 was able to enhance ALOX5 promoter activity by 47-fold, which was further stimulated when either vitamin D receptor and retinoid X receptor or SMAD3/SMAD4 were co-transfected. In addition, we investigated several histone deacetylase inhibitors (HDACi) in combination with gene knockdown experiments (HDAC1-3, MLL). We were able to demonstrate that a combined inhibition of HDAC1-3 induces ALOX5 promoter activity in an MLL-dependent manner. Surprisingly, a constitutive activation of ALOX5 by MLL-AF4 was inhibited by class I HDAC inhibitors, by relieving inhibitory functions deriving from MLL.Conversely, a knockdown of MLL increased the effects mediated by MLL-AF4. Thus, HDACi treatment seems to switch 'inactive MLL' into 'active MLL' and overwrites the dominant functions deriving from MLL-AF4.


Immunological effects of silica and asbestos.
Silicosis patients (SILs) and patients who have been exposed to asbestos develop not only respiratory diseases but also certain immunological disorders. In particular, SIL sometimes complicates autoimmune diseases such as systemic scleroderma, rheumatoid arthritis (known as Caplan syndrome), and systemic lupus erythematoses. In addition, malignant complications such as lung cancer and malignant mesothelioma often occur in patients exposed to asbestos, and may be involved in the reduction of tumor immunity. Although silica-induced disorders of autoimmunity have been explained as adjuvant-type effects of silica, more precise analyses are needed and should reflect the recent progress in immunomolecular findings. A brief summary of our investigations related to the immunological effects of silica/asbestos is presented. Recent advances in immunomolecular studies led to detailed analyses of the immunological effects of asbestos and silica. Both affect immuno-competent cells and these effects may be associated with the pathophysiological development of complications in silicosis and asbestos-exposed patients such as the occurrence of autoimmune disorders and malignant tumors, respectively. In addition, immunological analyses may lead to the development of new clinical tools for the modification of the pathophysiological aspects of diseases such as the regulation of autoimmunity or tumor immunity using cell-mediated therapies, various cytokines, and molecule-targeting therapies. In particular, as the incidence of asbestos-related malignancies is increasing and such malignancies have been a medical and social problem since the summer of 2005 in Japan, efforts should be focused on developing a cure for these diseases to eliminate nationwide anxiety.


One-pot synthesis of glutathione-responsive amphiphilic drug self-delivery micelles of doxorubicin-disulfide-methoxy polyethylene glycol for tumor therapy.
We present a novel glutathione-responsive amphiphilic drug self-delivery (DSD) micelle with one-pot synthesis to synergistically address the problems of controlled drug release, degradability, drug tracing and in vivo accumulated toxicity. The anticancer drug doxorubicin (DOX), disulfide-based diacrylate (DSDA) and amino-polyethylene glycol monomethyl ether were linked by Michael addition in one-pot synthesis. The accumulative release rate of DOX analogues with drug activity from the micelles was 67.9% under pH 7.4 and GSH = 1 mg mL-1 conditions after 72 h. The cell uptake experiment showed that the micelles of DOX-DSDA-PEG were indeed taken up by A549 cells and distributed to cell nuclei. The in vitro cell viability of A549 cells was evaluated by CCK-8 and Muse Annexin V & Dead Cell Kit. The results illustrated that the completely biodegradable micelles with glutathione-responsive bonds in the backbone are an effective drug self-delivery system for tumor therapy in the future.


MRI Shows that Exhaustion Syndrome Due to Chronic Occupational Stress is Associated with Partially Reversible Cerebral Changes.
The present study investigates the cerebral effects of chronic occupational stress and its possible reversibility. Forty-eight patients with occupational exhaustion syndrome (29 women) and 80 controls (47 women) underwent structural magnetic resonance imaging (MRI) and neuropsychological testing. Forty-four participants (25 patients, 19 controls) also completed a second MRI scan after 1-2 years. Only patients received cognitive therapy. The stressed group at intake had reduced thickness in the right prefrontal cortex (PFC) and left superior temporal gyrus (STG), enlarged amygdala volumes, and reduced caudate volumes. Except for the caudate volume, these abnormalities were more pronounced in females. They were all related to perceived stress, which was similar for both genders. Thickness of the PFC also correlated with an impaired ability to down-modulate negative emotions. Thinning of PFC and reduction of caudate volume normalized in the follow-up. The amygdala enlargement and the left STG thinning remained. Longitudinal changes were not detected among controls. Chronic occupational stress was associated with partially reversible structural abnormalities in key regions for stress processing. These changes were dynamically correlated with the degree of perceived stress, highlighting a possible causal link. They seem more pronounced in women, and could be a substrate for an increased cerebral vulnerability to stress-related psychiatric disorders.


Proteomics for the diagnosis of thyroid lesions: preliminary report.
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) is a unique proteomic technology that explores the spatial distribution of biomolecules directly in situ, thus integrating molecular and morphological information. The possibility of correlating distribution maps of multiple analyses with cytological features makes it an ideal research tool for discovering new diagnostic markers. A previous study showed that MALDI-IMS could help discrimination between different types of thyroid lesions, especially papillary thyroid carcinoma (PTC); the present feasibility study on ex vivo fine needle aspiration (FNA) smears describes its potential in detecting new proteomic targets of other thyroid lesions (follicular lesions, medullary carcinoma). MALDI-IMS was conducted on ex vivo FNAs obtained from surgical specimens and corresponding in vivo samples. Differences between proteomic profiles of different thyroid lesions were compared. Comparing the protein profiles of hyperplastic nodules obtained from three different patients with each other, and with a new PTC, showed a high degree of concordance, indicating good reproducibility of the IMS technology on cytological samples, suggesting its potential as a tool for biomarker discovery. Furthermore, comparison of the average proteomic profiles of hyperplastic nodules with a Hürthle cell adenoma revealed significant differences, underlying the capability of MALDI-IMS to distinguish between different thyroid lesions. Finally, the proteomic profile of medullary thyroid carcinoma was also characterized. Our results confirmed the possible role of MALDI-IMS in the search for diagnostic targets of PTC and follicular lesions, which could be applied in larger trials aimed at the identification of proteins, convertible to cost-effective diagnostic tools such as immunohistochemistry. These tests could be used to analyse in vivo cytological smears, improving the preoperative diagnosis of indeterminate thyroid nodules.


Genetics of type 1A (immune mediated) diabetes.
Type 1A (immune mediated) diabetes is genetically heterogeneous with important examples for man and animal models with major mutations (autosomal recessive and X-linked recessive) identified as well as oligogenic/polygenic inheritance. For the most common forms of type 1A diabetes alleles of DQ and DR within the major histocompatibility complex are important determinants of disease and allow identification of high risk individuals at birth. Further understanding of both common and rare genetic determinants of type 1A diabetes will contribute to understanding the pathogenesis of diabetes and of autoimmunity.


Evaluation of frontal plane QRS-T angle in patients with slow coronary flow.
Objectives. Frontal plane QRS-T angle is a novel marker of myocardial repolarization, and an increased frontal plane QRS-T angle is associated with adverse cardiac outcomes. Slow coronary flow may cause fatal cardiac arrhythmias by causing electrical abnormalities and altering ventricular repolarization. We aimed to evaluate the frontal plane QRS-T angle in patients with slow coronary flow. Design. A total of 60 consecutive patients with slow coronary flow and 60 consecutive patients with normal coronary flow were enrolled into the study. Laboratory and some electrocardiography parameters including frontal plane QRS-T angle were compared between the two groups. Results. We have found that the angle of frontal QRS-T was higher in the group with slow coronary flow (p < .001). In addition, there was a negative correlation between thrombolysis in myocardial infarction frame count and frontal QRS-T angle (r = -0.496, p < .001). Conclusions. In conclusion, increased frontal plane QRS-T angle might be an important indicator of slow coronary flow.


Comparison of serum, salivary, and rapid whole blood diagnostic tests for Helicobacter pylori and their validation against endoscopy based tests.
A rapid, reliable, and accurate test for the diagnosis of infection with Helicobacter pylori is needed for screening dyspeptic patients before referral for endoscopy. To compare a new rapid whole blood test (Helisal rapid blood, Cortecs), two serum enzyme linked immunosorbent assays (ELISAs; Helico-G, Shield and Helisal serum, Cortecs), and a salivary assay (Helisal saliva, Cortecs), with slide biopsy urease, 13C-urea breath test, and histology. Three hundred and three consecutive dyspeptic patients attending for gastroscopy underwent two antral biopsies for histology, and one for rapid slide biopsy urease test for assessment of H pylori status. Blood and saliva were also collected. One hundred of the patients also underwent a 13C-urea breath test. Gold standard positives were defined as those with at least two positive tests among slide urease, breath test, or histology, and gold standard negatives as those with all these (or two when the breath test was not done) negative. Of 300 patients (median age 63, range 28-89) eligible for analysis, 137 (46%) were gold standard positives, of which Helisal rapid blood identified 116, Helico-G 129, Helisal serum 130, and Helisal saliva 120; 137 (46%) were gold standard negatives of which the number falsely identified as positive was 30 by Helisal rapid blood, 45 by Helico-G, 41 by Helisal serum, and 41 by Helisal saliva. Sensitivities and specificities were: for the whole blood test 85% and 78% respectively; for Helico-G 94% and 67%, for Helisal serum 95% and 70%, and for Helisal saliva 84% and 70%. If endoscopy had been undertaken only on patients with positive tests two of 16 duodenal ulcers would have been missed if the Helisal rapid blood test was used, and one if any of the ELISA tests were used. None of the blood tests would have missed any of six gastric ulcers, but the salivary test would have missed one.


Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines.
Previous data from our laboratory has indicated that there is a functional link between the beta-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. GIRK1-4 expression and levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. GIRK protein levels were determined by western blots and cell proliferation was determined by a 5-bromo-2'-deoxyuridine (BrdU) assay. GIRK1 mRNA was expressed in three of six small cell lung cancer (SCLC) cell lines, and either GIRK2, 3 or 4 mRNA expression was detected in all six SCLC cell lines. Treatment of NCI-H69 with beta2-adrenergic antagonist ICI 118,551 (100 microM) daily for seven days led to slight decreases of GIRK1 mRNA expression levels. Treatment of NCI-H69 with the beta-adrenergic agonist isoproterenol (10 microM) decreased growth rates in these cells. The GIRK inhibitor U50488H (2 microM) also inhibited proliferation, and this decrease was potentiated by isoproterenol. In the SCLC cell lines that demonstrated GIRK1 mRNA expression, we also saw GIRK1 protein expression. We feel these may be important regulatory pathways since no expression of mRNA of the GIRK channels (1 & 2) was found in hamster pulmonary neuroendocrine cells, a suggested cell of origin for SCLC, nor was GIRK1 or 2 expression found in human small airway epithelial cells. GIRK (1,2,3,4) mRNA expression was also seen in A549 adenocarcinoma and NCI-H727 carcinoid cell lines. GIRK1 mRNA expression was not found in tissue samples from adenocarcinoma or squamous cancer patients, nor was it found in NCI-H322 or NCI-H441 adenocarcinoma cell lines. GIRK (1,3,4) mRNA expression was seen in three squamous cell lines, GIRK2 was only expressed in one squamous cell line. However, GIRK1 protein expression was not seen in any non-SCLC cells. We feel that this data may indicate that stimulation of GIRK1 or GIRK2 channels may be important in lung cancer. Stimulation of GIRK channels and beta-adrenergic signaling may activate similar signaling pathways in both SCLC and breast cancer, but lead to different results.


Multicenter Evaluation of the Fully Automated PCR-Based Idylla EGFR Mutation Assay on Formalin-Fixed, Paraffin-Embedded Tissue of Human Lung Cancer.
Before initiating treatment of advanced non-small-cell lung cancer with tyrosine kinase inhibitors (eg, erlotinib, gefitinib, osimertinib, and afatinib), which inhibit the catalytic activity of epidermal growth factor receptor (EGFR), clinical guidelines require determining the EGFR mutational status for activating (EGFR exons 18, 19, 20, or 21) and resistance (EGFR exon 20) mutations. The EGFR resistance mutation T790M should be monitored at cancer progression. The Idylla EGFR Mutation Assay, performed on the Idylla molecular diagnostics platform, is a fully automated (<2.5 hours turnaround time) sample-to-result molecular test to qualitatively detect 51 EGFR oncogene point mutations, deletions, or insertions. In a 15-center evaluation, Idylla results on 449 archived formalin-fixed, paraffin-embedded tissue sections, originating from non-small-cell lung cancer biopsies and resection specimens, were compared with data obtained earlier with routine reference methods, including next-generation sequencing, Sanger sequencing, pyrosequencing, mass spectrometry, and PCR-based assays. When results were discordant, a third method of analysis was performed, when possible, to confirm test results. After confirmation testing and excluding invalids/errors and discordant results by design, a concordance of 97.6% was obtained between Idylla and routine test results. Even with <10 mm2 of tissue area, a valid Idylla result was obtained in 98.9% of the cases. The Idylla EGFR Mutation Assay enables sensitive detection of most relevant EGFR mutations in concordance with current guidelines, with minimal molecular expertise or infrastructure.


The level of quality of radiology services in Turkey: a sampling analysis.
To determine the quality of radiographs, which have been referred from 40 different institutions for consultation, to discuss the causes of wasted resources, and to present possible solutions. Five experienced radiology instructors determined the types of radiological examinations referred for consultation (conventional radiography, mammography, computed tomography and magnetic resonance imaging), the institutions at which they were performed (university or state hospital, private health center), and assessed the coverage area, field of vision (FOV), and dosage of x-ray. They also investigated problems in film processing, defects in sequence-printing windows, checked window levels, and checked the amount and timing of contrast material used. According to these criteria, the reviewers subjectively classified each radiograph as: 1. Poor, examination should be completely repeated, 2. Fair, examination should be partially repeated, 3. Good, accepted as adequate, no need for an additional examination, 4. Excellent, examination was as it should be. We reviewed 120 radiological examinations from 40 different institutions in 4 reference centers. Frequency of problems determined for each category was as follows: coverage area 32.5% (39/120), FOV 16% (14/86), X-ray dosage 16% (15/94), film processing 31% (37/120), sequence or window 65% (53/81), window level 44% (36/81); contrast material 51% (25/49), timing of contrast material 61% (30/49). Only 22% of the examinations were classified as excellent, whereas 47% required complete or partial repetition. Approximately half of the radiological examinations in our sampling required partial or complete repetition. Health, ethical, and economic aspects of the problem necessitates the prompt application of measures to establish radiological quality control and standardization procedures.


Effects of transcranial direct current stimulation upon attention and visuoperceptual function in Lewy body dementia: a preliminary study.
Individuals with Lewy body dementia (LBD) typically exhibit impairments in attentional and executive function. Current pharmacological treatments have limited efficacy, with associated side effects. Transcranial direct current stimulation (tDCS) may represent an alternative treatment, as cognitive improvements have been demonstrated in healthy individuals. However, no studies to date have assessed the feasibility of tDCS in an LBD population. The aim of this preliminary study, therefore, was to assess the tolerability of tDCS, as well as its effects upon attentional and visuoperceptual performance, in LBD patients. Thirteen participants completed attentional (simple reaction time, choice reaction time, and digit vigilance) and forced-choice visuoperceptual (angle and motion perception) tasks before and after one 20-min session of active tDCS (0.08 mA/cm2). The anodal electrode was applied to the left dorsolateral prefrontal cortex and the cathodal electrode was applied to the right deltoid. Attentional (task accuracy and reaction time to correct answers) and visuoperceptual (task accuracy and difficulty) outcome measures were compared using paired t-tests. All participants tolerated stimulation and did not report any side effects during or immediately after stimulation. Post-stimulation improvements were observed in the choice reaction time (increased percentage of correct answers; p = 0.01) and digit vigilance (reduced mean reaction time to correct answers; p = 0.02) attention tasks. Visuoperceptual task performance did not improve (all p-values > 0.05). Attentional, but not visuoperceptual, improvements were observed following stimulation in LBD patients. Larger-scale, placebo-controlled trials are needed to confirm whether tDCS is a useful treatment option for attentional deficits in LBD.


Uncoupling of IL-6 signaling and LC3-associated phagocytosis drives immunoparalysis during sepsis.
Immune deactivation of phagocytes is a central event in the pathogenesis of sepsis. Herein, we identify a master regulatory role of IL-6 signaling on LC3-associated phagocytosis (LAP) and reveal that uncoupling of these two processes during sepsis induces immunoparalysis in monocytes/macrophages. In particular, we demonstrate that activation of LAP by the human fungal pathogen Aspergillus fumigatus depends on ERK1/2-mediated phosphorylation of p47phox subunit of NADPH oxidase. Physiologically, autocrine IL-6/JAK2/Ninein axis orchestrates microtubule organization and dynamics regulating ERK recruitment to the phagosome and LC3+ phagosome (LAPosome) formation. In sepsis, loss of IL-6 signaling specifically abrogates microtubule-mediated trafficking of ERK, leading to defective activation of LAP and impaired killing of bacterial and fungal pathogens by monocytes/macrophages, which can be selectively restored by IL-6 supplementation. Our work uncovers a molecular pathway linking IL-6 signaling with LAP and provides insight into the mechanisms underlying immunoparalysis in sepsis.


Cellular functions of phosphatidylinositol 3-phosphate and FYVE domain proteins.
PtdIns3P is a phosphoinositide 3-kinase product that has been strongly implicated in regulating membrane trafficking in both mammalian and yeast cells. PtdIns3P has been shown to be specifically located on membranes associated with the endocytic pathway. Proteins that contain FYVE zinc-finger domains are recruited to PtdIns3P-containing membranes. Structural information is now available concerning the interaction between FYVE domains and PtdIns3P. A number of proteins have been identified which contain a FYVE domain, and in this review we discuss the functions of PtdIns3P and its FYVE-domain-containing effector proteins in membrane trafficking, cytoskeletal regulation and receptor signalling.


Simulating protein motions with rigidity analysis.
Protein motions, ranging from molecular flexibility to large-scale conformational change, play an essential role in many biochemical processes. Despite the explosion in our knowledge of structural and functional data, our understanding of protein movement is still very limited. In previous work, we developed and validated a motion planning based method for mapping protein folding pathways from unstructured conformations to the native state. In this paper, we propose a novel method based on rigidity theory to sample conformation space more effectively, and we describe extensions of our framework to automate the process and to map transitions between specified conformations. Our results show that these additions both improve the accuracy of our maps and enable us to study a broader range of motions for larger proteins. For example, we show that rigidity-based sampling results in maps that capture subtle folding differences between protein G and its mutants, NuG1 and NuG2, and we illustrate how our technique can be used to study large-scale conformational changes in calmodulin, a 148 residue signaling protein known to undergo conformational changes when binding to Ca(2+). Finally, we announce our web-based protein folding server which includes a publicly available archive of protein motions: (http://parasol.tamu.edu/foldingserver/).


Observation of crystalline changes of titanium dioxide during lithium insertion by visible spectrum analysis.
Real-time analysis of changes in the atomic environment of materials is a cutting edge technology that is being used to explain reaction dynamics in many fields of science. Previously, this kind of analysis was only possible using heavy nucleonic equipment such as XANES and EXAFS, or Raman spectroscopy on a moderate scale. Here, a new methodology is described that can be used to track changes in crystalline developments during complex Li insertion reactions via the observation of structural color. To be specific, the changes in atomic crystalline and nanostructure are shown during Li insertion in a complex TiO2 polymorph. Structural color corresponds to the refractive indices of materials originating from their atomic bonding nature and precise wave interferences in accordance with their nanostructure. Therefore, this new analysis simultaneously reveals changes in the nanostructure as well as changes in the atomic bonding nature of materials.


Advanced provision of emergency contraception does not reduce abortion rates.
A number of small studies have demonstrated increased use of emergency contraception (EC) when women have a supply available at home. It has been suggested that widespread use of EC could reduce abortion rates. We undertook a community intervention study designed to determine whether offering advanced supplies of EC to large numbers of women influenced abortion rates. All women aged between 16 and 29 years living in Lothian, Scotland, were offered, through health services, five courses of EC without cost to keep at home. Of a population of around 85,000 women in this age group, the study showed that an estimated 17,800 women took a supply of EC home and over 4500 of them gave at least one course to a friend. It was found that nearly half (45%) of women who had a supply used at least one course during the 28 months that the study lasted. In total, an estimated 8081 courses of EC were used. EC was used within 24 h after intercourse on 75% of occasions. Abortion rates in Lothian were compared with those from three other health board areas of Scotland. No effect on abortion rates was demonstrated with advanced provision of EC. The results of this study suggest that widespread distribution of advanced supplies of EC through health services may not be an effective way to reduce the incidence of unintended pregnancy in the UK.


Application of a Rank-Based Method for Improved Cultivar Selection in Soft Red Winter Wheat.
Both grain yield and disease performance are important factors to consider for winter wheat (Triticum aestivum) cultivar selection. However, disease severity and yield data are often presented separately, making it difficult to compare values across multiple environments. The objective of this study was to use a rank-based method to compare cultivars based on yield and disease performance combined across multiple environments. Thirty-six wheat cultivars were planted at each of four Wisconsin locations (Arlington, Chilton, Janesville, and Lancaster) in 2009 and 2010. Plots were assessed four times during the growing season for powdery mildew, Septoria/Stagonospora leaf blotch, and leaf rust. Incidence and severity of Fusarium head blight were assessed at Zadoks growth stage 85 (soft dough). Within each location-year, cultivars were ranked for severity of each disease and for grain yield. One-way analysis of variance was performed to calculate an overall rank value that incorporated data for all four diseases and yield across the eight location-years. There was an effect of cultivar on overall rank (P < 0.0001). Powdery mildew rank was strongly correlated with overall rank (Spearman's rho = 0.485, P = 0.005), as was yield rank (Spearman's rho = 0.674, P < 0.0001). Cultivars described as "best" or "worst" cultivars were generally more consistent in their rankings across different measures. The use of a rank-based method provides an approach that will allow growers to base cultivar selection on multiple performance measures across multiple environments.


Genome-wide analysis of the Glycerol-3-Phosphate Acyltransferase (GPAT) gene family reveals the evolution and diversification of plant GPATs.
sn-Glycerol-3-phosphate 1-O-acyltransferase (GPAT) is an important enzyme that catalyzes the transfer of an acyl group from acyl-CoA or acyl-ACP to the sn-1 or sn-2 position of sn-glycerol-3-phosphate (G3P) to generate lysophosphatidic acids (LPAs). The functional studies of GPAT in plants demonstrated its importance in controlling storage and membrane lipid. Identifying genes encoding GPAT in a variety of plant species is crucial to understand their involvement in different metabolic pathways and physiological functions. Here, we performed genome-wide and evolutionary analyses of GPATs in plants. GPAT genes were identified in all algae and plants studied. The phylogenetic analysis showed that these genes group into three main clades. While clades I (GPAT9) and II (soluble GPAT) include GPATs from algae and plants, clade III (GPAT1-8) includes GPATs specific from plants that are involved in the biosynthesis of cutin or suberin. Gene organization and the expression pattern of GPATs in plants corroborate with clade formation in the phylogeny, suggesting that the evolutionary patterns is reflected in their functionality. Overall, our results provide important insights into the evolution of the plant GPATs and allowed us to explore the evolutionary mechanism underlying the functional diversification among these genes.


Protection of C. elegans from anoxia by HYL-2 ceramide synthase.
Oxygen deprivation is rapidly deleterious for most organisms. However, Caenorhabditis elegans has developed the ability to survive anoxia for at least 48 hours. Mutations in the DAF-2/DAF-16 insulin-like signaling pathway promote such survival. We describe a pathway involving the HYL-2 ceramide synthase that acts independently of DAF-2. Loss of the ceramide synthase gene hyl-2 results in increased sensitivity of C. elegans to anoxia. C. elegans has two ceramide synthases, hyl-1 and hyl-2, that participate in ceramide biogenesis and affect its ability to survive anoxic conditions. In contrast to hyl-2(lf) mutants, hyl-1(lf) mutants are more resistant to anoxia than normal animals. HYL-1 and HYL-2 have complementary specificities for fatty acyl chains. These data indicate that specific ceramides produced by HYL-2 confer resistance to anoxia.


Pulmonary Hypertension and Polycythemia Secondary to Pulmonary Alveolar Microlithiasis Treated with Sequential Bilateral Lung Transplant: A Case Study and Literature Review.
BACKGROUND Pulmonary alveolar microlithiasis is an autosomal recessive disease in which a mutation in the SLC34A2 gene that codes for a sodium phosphate type IIb transporter protein (expressed in human epithelial tissues and functions in the clearance of phosphate ions) leads to the formation of extensive pulmonary intra-alveolar microliths. The subsequent characteristic clinical features of dyspnea and hypoxia are a manifestation of these microliths. There have been fewer than 1000 cases of pulmonary alveolar microlithiasis reported worldwide, and there have been 19 reported lung-transplanted patients. CASE REPORT A 49-year-old Saudi male patient presented with longstanding history of easy fatigability and tiredness on exertion since he was 16 years old. Throughout his follow-up in different hospitals (1986-1989), tuberculosis and pulmonary fibrosis were suspected. The patient was lost to follow-up between 1989 and 2001. In 2002, he presented to the emergency room with coughing, shortness of breath on exertion, abdominal swelling, and pedal edema. An investigation with chest x-rays, CT scan, electrocardiogram, and an echocardiogram was conducted. After referral to a tertiary care center, the patient was diagnosed with pulmonary alveolar microlithiasis. He subsequently developed pulmonary hypertension and polycythemia and therefore received a bilateral lung transplant in 2016. Following the lung transplant, he developed a mild reperfusion injury and tonic-clonic seizures, requiring ICU admission. After a successful extubatation with stable vitals and good recovery, he was discharged home in stable condition with planned follow-up. CONCLUSIONS We report a case of pulmonary alveolar microlithiasis successfully treated with a bilateral lung transplant. Although pulmonary alveolar microlithiasis is a rare entity, healthcare providers should consider it in the differential diagnoses of parenchymal lung diseases and differentiate it from tuberculosis and pulmonary fibrosis.


Sports injuries in an America's Cup yachting crew: A 4-year epidemiological study covering the 2007 challenge.
The aim of this study was to describe the injuries sustained by an America's Cup crew during eight preparatory competitions of the 32nd America's Cup 2007 and the Louis Vuitton Cup (from October 2004 to June 2007). The anatomical location, type of injury, and mechanism of injury were recorded. The injuries were categorized based on each sailor's position on the boat according to three intensities of physical demands. The injury rates per sailor and per 1000 h of competition were determined. In total, 90 injuries were registered. The overall incidence was 10 injuries per 1000 competition hours. Overuse injuries accounted for 76.6% of all lesions. The most common anatomical location of injuries was the upper limb (36.6%), followed by the upper dorsal and cervical spine (34.4%). Frequency of injury was related to the sailor's position on the boat, being higher in the group with more demanding activities (grinder, bowman, and mastman). Most injuries (67%) were sustained by this group of sailors. The most common injuries in this group were muscle contractures of the quadratus lumborum (11), trapezius (8), and rhomboid (7). There were eight cases of elbow epicondylitis, four cases of tendinopathy of the supraspinous tendon, and three cases of tendinopathy of the biceps brachii. An America's Cup yachting crew is exposed to a high risk of overuse injuries, especially those sailors whose boat position involves high-intensity activity.


Occupational therapy treatment goals for adults with developmental disabilities.
This study examined whether occupational therapists were using treatment time to prepare adults with developmental disabilities for independent living opportunities. Eighty-eight occupational therapy evaluations for 54 adults with developmental disabilities in 18 different community agencies in the Boston area were reviewed to ascertain the focus of treatment goals that reflect treatment approaches. Results indicate that remedial goals significantly outnumbered adaptive ones. Within the adaptive goals, self-care and community living skills were equally represented, but some important community skill areas like advocacy, money management, personal care attendant management, sexuality, and transportation received little or no attention. Implications of these findings for therapists, clients, and third-party payers are discussed.


An architectural framework that may lie at the core of the postsynaptic density.
The postsynaptic density (PSD) is a complex assembly of proteins associated with the postsynaptic membrane that organizes neurotransmitter receptors, signaling pathways, and regulatory elements within a cytoskeletal matrix. Here we show that the sterile alpha motif domain of rat Shank3/ProSAP2, a master scaffolding protein located deep within the PSD, can form large sheets composed of helical fibers stacked side by side. Zn2+, which is found in high concentrations in the PSD, binds tightly to Shank3 and may regulate assembly. Sheets of the Shank protein could form a platform for the construction of the PSD complex.


Alteration of seed fatty acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity.
In characterizing the enzymes involved in the formation of very long-chain fatty acids (VLCFAs) in the Brassicaceae, we have generated a series of mutants of Arabidopsis thaliana that have reduced VLCFA content. Here we report the characterization of a seed lipid mutant, AS11, which, in comparison to wild type (WT), has reduced levels of 20:1 and 18:1 and accumulates 18:3 as the major fatty acid in triacylglycerols. Proportions of 18:2 remain similar to WT. Genetic analyses indicate that the fatty acid phenotype is caused by a semidominant mutation in a single nuclear gene, designated TAG1, located on chromosome 2. Biochemical analyses have shown that the AS11 phenotype is not due to a deficiency in the capacity to elongate 18:1 or to an increase in the relative delta 15 or delta 12 desaturase activities. Indeed, the ratio of desaturase/elongase activities measured in vitro is virtually identical in developing WT and AS11 seed homogenates. Rather, the fatty acid phenotype of AS11 is the result of reduced diacylglycerol acyltransferase activity throughout development, such that triacylglycerol biosynthesis is reduced. This leads to a reduction in 20:1 biosynthesis during seed development, leaving more 18:1 available for desaturation. Thus, we have demonstrated that changes to triacylglycerol biosynthesis can result in dramatic changes in fatty acid composition and, in particular, in the accumulation of VLCFAs in seed storage lipids.


Evidence of Environmental Changes Caused by Chinese Island-Building.
This paper quantifies environmental effects of island-building operations in the South China Sea, which result from dredging and can negatively impact marine flora, fauna, and ecosystems. The extent of the damage caused by island-creation is believed to be large, as the South China Sea reefs support the largest concentration of marine biodiversity on Earth. Through use of satellite imagery, we investigate the island-construction on Mischief Reef in the South China Sea, showing backscatter increases of up to 350% in waters surrounding the reef, with plumes of excess sediment exceeding 250 km2 at times during island-construction, and the cumulative area impacted by dredging exceeding 1,200 km2. Comparison of satellite-derived chlorophyll-a, backscatter, absorption and remote sensing reflectance at 412 nm suggest that dredging activities led to a decrease in biological health of the region resulting from the smothering of natural benthic habitats and reef complexes with sediment. We anticipate this ex post facto quantification of the connectivity between island-construction, large particulate plumes and a decrease in absorption related to marine life in the water column to establish a starting point for further study into ecosystem impact. The potential associations between these damages and a long-term reduction in ocean life and resources could serve inter-governmental bodies with a baseline metric for evaluating the level of damage caused. This may result in both forward-looking deterrent policies that limit island-building as well as backward-looking compensation.


Different Doses and Routes of Administration of Methimazole Affect Thyroid Status in Methimazole-induced Hypothyroidism in Rats.
It is agreed that methimazole (MMI) can be administered to induce hypothyroidism. However, there are conflicting data about its effect on thyroid function and development in rats through different administrations. In the present study, we established and compared differences of the rat hypothyroid model induced by MMI added to drinking water or given through an intragastric tube. Sixty-four male Wistar rats were randomly divided into seven groups. Methimazole was added to the drinking water (0.025%, 0.04% or 0.1% wt/vol), or through intragastric gavage (5 mg/100 g body weight (bw) or 8 mg/100 g bw) one time each day for 21 days. The rats were weighed every seven days. Blood samples were taken in order to detect the concentrations of serum triiodothyronine (T3), thyroxine (T4) and thyrotropin (TSH) at the end of the experiments. Our results indicate that the effect of methimazole on a rat's thyroid function and body weight is similar in both the group given 0.1% concentration in drinking water and the group which received 8 mg/100 g bw once daily through the intragastric tube. Also, a similar effect was observed in the 0.025%, 0.04% and 5 mg/100 g bw groups. These findings suggest that a relationship between the concentration of MMI by oral administration and the dose of it through intragastric administration could exist, and may contribute to inducing hypothyroidism in rats.


Neuronal model with distributed delay: analysis and simulation study for gamma distribution memory kernel.
A single neuronal model incorporating distributed delay (memory)is proposed. The stochastic model has been formulated as a Stochastic Integro-Differential Equation (SIDE) which results in the underlying process being non-Markovian. A detailed analysis of the model when the distributed delay kernel has exponential form (weak delay) has been carried out. The selection of exponential kernel has enabled the transformation of the non-Markovian model to a Markovian model in an extended state space. For the study of First Passage Time (FPT) with exponential delay kernel, the model has been transformed to a system of coupled Stochastic Differential Equations (SDEs) in two-dimensional state space. Simulation studies of the SDEs provide insight into the effect of weak delay kernel on the Inter-Spike Interval(ISI) distribution. A measure based on Jensen-Shannon divergence is proposed which can be used to make a choice between two competing models viz. distributed delay model vis-á-vis LIF model. An interesting feature of the model is that the behavior of (CV(t))((ISI)) (Coefficient of Variation) of the ISI distribution with respect to memory kernel time constant parameter η reveals that neuron can switch from a bursting state to non-bursting state as the noise intensity parameter changes. The membrane potential exhibits decaying auto-correlation structure with or without damped oscillatory behavior depending on the choice of parameters. This behavior is in agreement with empirically observed pattern of spike count in a fixed time window. The power spectral density derived from the auto-correlation function is found to exhibit single and double peaks. The model is also examined for the case of strong delay with memory kernel having the form of Gamma distribution. In contrast to fast decay of damped oscillations of the ISI distribution for the model with weak delay kernel, the decay of damped oscillations is found to be slower for the model with strong delay kernel.


Spin-echo small-angle neutron scattering (SESANS) studies of diblock copolymer nanoparticles.
Poly(glycerol monomethacrylate)-poly(benzyl methacrylate) (PGMA-PBzMA) diblock copolymer nanoparticles were synthesized via polymerization-induced self-assembly (PISA) using reversible addition-fragmentation chain-transfer (RAFT) aqueous emulsion polymerization in D2O. Such PISA syntheses produce sterically-stabilized nanoparticles in situ and can be performed at relatively high copolymer concentrations (up to 50 wt%). This PGMA-PBzMA formulation is known to form only spherical nanoparticles in water using aqueous emulsion polymerization (Macromolecules, 2014, 47, 5613-5623), which makes it an ideal model system for exploring new characterization methods. The polymer micelles were characterized using small-angle X-ray scattering (SAXS) and a recently developed form of neutron scattering, spin-echo small-angle neutron scattering (SESANS). As far as we are aware, this is the first report of a study of polymer micelles by SESANS, and the data agree well with reciprocal-space scattering. Using this technique enables characterization of the concentrated, as synthesized dispersions directly without dilution, and this will provide a method to study self-assembled polymer systems that have concentration dependent morphologies, while still maintaining the advantages of scattering techniques.


Interaural time-delay sensitivity in bilateral cochlear implant users: effects of pulse rate, modulation rate, and place of stimulation.
Electrical interaural time delay (ITD) discrimination was measured using 300-ms bursts applied to binaural pitch matched electrodes at basal, mid, and apical locations in each ear. Six bilateral implant users, who had previously shown good ITD sensitivity at a pulse rate of 100 pulses per second (pps), were assessed. Thresholds were measured as a function of pulse rate between 100 and 1,000 Hz, as well as modulation rate over that same range for high-rate pulse trains at 6,000 pps. Results were similar for all three places of stimulation and showed decreasing ITD sensitivity as either pulse rate or modulation rate increased, although the extent of that effect varied across subjects. The results support a model comprising a common ITD mechanism for high- and low-frequency places of stimulation, which, for electrical stimulation, is rate-limited in the same way across electrodes because peripheral temporal responses are largely place invariant. Overall, ITD sensitivity was somewhat better with unmodulated pulse trains than with high-rate pulse trains modulated at matched rates, although comparisons at individual rates showed that difference to be significant only at 300 Hz. Electrodes presenting with the lowest thresholds at 600 Hz were further assessed using bursts with a ramped onset of 10 ms. The slower rise time resulted in decreased performance in four of the listeners, but not in the two best performers, indicating that those two could use ongoing cues at 600 Hz. Performance at each place was also measured using single-pulse stimuli. Comparison of those data with the unmodulated 300-ms burst thresholds showed that on average, the addition of ongoing cues beyond the onset enhanced overall ITD sensitivity at 100 and 300 Hz, but not at 600 Hz. At 1,000 Hz, the added ongoing cues actually decreased performance. That result is attributed to the introduction of ambiguous cues within the physiologically relevant range and increased dichotic firing.


Development and validation of a deep-learning-based pediatric early warning system: A single-center study.
Early detection and prompt intervention for clinically deteriorating events are needed to improve clinical outcomes. There have been several attempts at this, including the introduction of rapid response teams (RRTs) with early warning scores. We developed a deep-learning-based pediatric early warning system (pDEWS) and validated its performance. This single-center retrospective observational cohort study reviewed, 50,019 pediatric patients admitted to the general ward in a tertiary-care academic children's hospital from January 2012 to December 2018. They were split by admission date into a derivation and a validation cohort. We developed a pDEWS for the early prediction of cardiopulmonary arrest and unexpected ward-to-pediatric intensive care unit (PICU) transfer. Then, we validated this system by comparing modified pediatric early warning score (PEWS), random forest (RF); an ensemble model of multiple decision trees and logistic regression (LR); a statistical model that uses a logistic function. For predicting cardiopulmonary arrest, the pDEWS (area under the receiver operating characteristic curve (AUROC), 0.923) outperformed modified PEWS (AUROC, 0.769) and reduced the mean alarm count per day (MACPD) and number needed to examine (NNE) by 82.0% (from 46.7 to 8.4 MACPD) and 89.5% (from 0.303 to 0.807), respectively. Furthermore, for predicting unexpected ward-to-PICU transfer pDEWS also showed superior performance compared to existing methods. Our study showed that pDEWS was superior to the modified PEWS and prediction models using RF and LR. This study demonstrates that the integration of the pDEWS into RRTs could increase operational efficiency and improve clinical outcomes.


A short-term clinical study of marginal bone level change around microthreaded and platform-switched implants.
The marginal bone levels around implants following restoration are used as a reference for evaluating implant success and survival. Two design concepts that can reduce crestal bone resorption are the microthread and platform-switching concepts. The aims of this study were to analyze the placement of microthreaded and platform-switched implants and their short-term survival rate, as well as the level of bone around the implants. The subjects of this study were 27 patients (79 implants) undergoing treatment with microthreaded and platform-switched implants between October 2008 and July 2009 in the Dental Hospital of Yonsei University Department of Periodontology. The patients received follow-up care more than 6 months after the final setting of the prosthesis, at which time periapical radiographs were taken. The marginal bone level was measured from the reference point to the lowest observed point of contact between the marginal bone and the fixture. Comparisons were made between radiographs taken at the time of fixture installation and those taken at the follow-up visit. During the study period (average of 11.8 months after fixture installation and 7.4 months after the prosthesis delivery), the short-term survival rate of microthreaded and platform-switched implants was 100% and the marginal bone loss around implants was 0.16±0.08 mm, the latter of which is lower than the previously reported values. This short-term clinical study has demonstrated the successful survival rates of a microthread and platform-switched implant system, and that this system is associated with reduced marginal bone loss.


Helicobacter pylori-associated ammonia production enhances neutrophil-dependent gastric mucosal cell injury.
The role of neutrophil and its chlorinated oxidant were investigated in Helicobacter pylori-induced gastric mucosal injury in vitro. Luminol-dependent chemiluminescence (ChL) was used to detect neutrophil-derived oxidants. ChL activity was significantly elevated when neutrophils were incubated in H. pylori, indicating that H. pylori actually elicits oxidative burst of neutrophils. To assess whether H. pylori-activated neutrophils exert the cytotoxicity for gastric mucosal cells, rabbit gastric mucosal cell was monolayered in culture wells and labeled with a fluorescence dye, 2',7'-bis(2-carboxyethyl)-5(6)carboxy-fluorescein, which is retained in the intracellular space as long as the cell membrane is intact. Labeled cells were coincubated with neutrophils and H. pylori. We inferred from the cytotoxicity index (specific %cytotoxicity), which was calculated from fluorometrical measurements of supernatant and lysate, that the mucosal cells were significantly damaged by H. pylori-activated neutrophils. This injury was largely attenuated by eliminating urea from the incubation mixture or by acetohydroxamic acid, a potent urease inhibitor. Additionally, the scavengers of neutrophil-derived oxidants, including taurine, methionine, and catalase, also attenuated this injury. Cultured mucosal cells that were exposed to the solution containing monochloramine (an oxidant yielded by reaction of hypochlorous acid and ammonia) were highly damaged compared with cells exposed to hypochlorous acid or hydrogen peroxide at physiological concentrations. These data suggest that H. pylori-activated neutrophils promote gastric mucosal cell injury and that monochloramine plays a unique and important role in this process.


Identifying targets of the rough homeobox gene of Drosophila: evidence that rhomboid functions in eye development.
In order to identify potential target genes of the rough homeodomain protein, which is known to specify some aspects of the R2/R5 photoreceptor subtype in the Drosophila eye, we have carried out a search for enhancer trap lines whose expression is rough-dependent. We crossed 101 enhancer traps that are expressed in the developing eye into a rough mutant background, and have identified seven lines that have altered expression patterns. One of these putative rough target genes is rhomboid, a gene known to be required for dorsoventral patterning and development of some of the nervous system in the embryo. We have examined the role of rhomboid in eye development and find that, while mutant clones have only a subtle phenotype, ectopic expression of the gene causes the non-neuronal mystery cells to be transformed into photoreceptors. We propose that rhomboid is a part of a partially redundant network of genes that specify photoreceptor cell fate.


High spatial resolution brain functional MRI using submillimeter balanced steady-state free precession acquisition.
One of the technical advantages of functional magnetic resonance imaging (fMRI) is its precise localization of changes from neuronal activities. While current practice of fMRI acquisition at voxel size around 3 × 3 × 3 mm(3) achieves satisfactory results in studies of basic brain functions, higher spatial resolution is required in order to resolve finer cortical structures. This study investigated spatial resolution effects on brain fMRI experiments using balanced steady-state free precession (bSSFP) imaging with 0.37 mm(3) voxel volume at 3.0 T. In fMRI experiments, full and unilateral visual field 5 Hz flashing checkerboard stimulations were given to healthy subjects. The bSSFP imaging experiments were performed at three different frequency offsets to widen the coverage, with functional activations in the primary visual cortex analyzed using the general linear model. Variations of the spatial resolution were achieved by removing outer k-space data components. Results show that a reduction in voxel volume from 3.44 × 3.44 × 2 mm(3) to 0.43 × 0.43 × 2 mm(3) has resulted in an increase of the functional activation signals from (7.7 ± 1.7)% to (20.9 ± 2.0)% at 3.0 T, despite of the threefold SNR decreases in the original images, leading to nearly invariant functional contrast-to-noise ratios (fCNR) even at high spatial resolution. Activation signals aligning nicely with gray matter sulci at high spatial resolution would, on the other hand, have possibly been mistaken as noise at low spatial resolution. It is concluded that the bSSFP sequence is a plausible technique for fMRI investigations at submillimeter voxel widths without compromising fCNR. The reduction of partial volume averaging with nonactivated brain tissues to retain fCNR is uniquely suitable for high spatial resolution applications such as the resolving of columnar organization in the brain.


Expression of the c-myc oncogene and the presence of HPV 18: possible surrogate markers for cervical cancer?
The study aims to evaluate the cause of cervical cancer in a cohort of patients and to establish whether or not human papillomavirus (HPV) is the leading risk factor and to determine whether or not c-myc oncogene over-expression is a predicative marker for the disease. Cone biopsy samples are examined from 53 patients diagnosed with either adenocarcinoma or squamous cell carcinoma of the cervix. Results showed that 19% of the patients studied were positive for high-grade HPV 18 DNA by polymerase chain reaction (PCR). For the c-myc gene expression, only three (23%) of the 13 control slides were positive. Of 49 known cervical cancer patients examined, 41% were positive, 51% were negative and 8% were doubtful. Of those who were positive for HPV, only two were positive for a mutation in the c-myc gene and one slide gave a doubtful result. P value for hysterectomy patients was 0.23 and for cancer patients was 0.48. In the cervical cancer patients studied, the HPV 18 prevalence rate was very low compared to that found in other studies. Therefore, the presence of HPV and expression of the c-myc oncogene cannot be used as surrogate markers for cervical cancer.


Neurotensin Induces Presynaptic Depression of D2 Dopamine Autoreceptor-Mediated Neurotransmission in Midbrain Dopaminergic Neurons.
Increased dopaminergic signaling is a hallmark of severe mesencephalic pathologies such as schizophrenia and psychostimulant abuse. Activity of midbrain dopaminergic neurons is under strict control of inhibitory D2 autoreceptors. Application of the modulatory peptide neurotensin (NT) to midbrain dopaminergic neurons transiently increases activity by decreasing D2 dopamine autoreceptor function, yet little is known about the mechanisms that underlie long-lasting effects. Here, we performed patch-clamp electrophysiology and fast-scan cyclic voltammetry in mouse brain slices to determine the effects of NT on dopamine autoreceptor-mediated neurotransmission. Application of the active peptide fragment NT8-13 produced synaptic depression that exhibited short- and long-term components. Sustained depression of D2 autoreceptor signaling required activation of the type 2 NT receptor and the protein phosphatase calcineurin. NT application increased paired-pulse ratios and decreased extracellular levels of somatodendritic dopamine, consistent with a decrease in presynaptic dopamine release. Surprisingly, we observed that electrically induced long-term depression of dopaminergic neurotransmission that we reported previously was also dependent on type 2 NT receptors and calcineurin. Because electrically induced depression, but not NT-induced depression, was blocked by postsynaptic calcium chelation, our findings suggest that endogenous NT may act through a local circuit to decrease presynaptic dopamine release. The current research provides a mechanism through which augmented NT release can produce a long-lasting increase in membrane excitability of midbrain dopamine neurons. Whereas plasticity of glutamate synapses in the brain has been studied extensively, demonstrations of plasticity at dopaminergic synapses have been more elusive. By quantifying inhibitory neurotransmission between midbrain dopaminergic neurons in brain slices from mice we have discovered that the modulatory peptide neurotensin can induce a persistent synaptic depression by decreasing dopamine release. This depression of inhibitory synaptic input would be expected to increase excitability of dopaminergic neurons. Induction of the plasticity can be pharmacologically blocked by antagonists of either the protein phosphatase calcineurin or neurotensin receptors, and persists surprisingly long after a brief exposure to the peptide. Since neurotensin-dopamine interactions have been implicated in hyperdopaminergic pathologies, these findings describe a synaptic mechanism that could contribute to addiction and/or schizophrenia.


Ventriculoatrial conduction in patients without high-grade AV block: when is it present?
Ventriculoatrial (VA) conduction is a critical component in many arrhythmias, has a diagnostic value in electrophysiology study (EPS), and is implicated in pacemaker-mediated arrhythmias. This study sought to characterize retrograde conduction during EPS and to utilize it as a diagnostic tool in patients without AV block. Patients with intact AV conduction undergoing EPS were included in this study to systematically evaluate baseline VA conduction. If absent, parahisian pacing was used to determine the level of block (nodal or infranodal). Recovery of VA conduction with increased sympathetic activity was assessed with isoproterenol infusion. Baseline characteristics and electrophysiological data were collected and analyzed. Among the 801 patients studied, VA conduction was present in 98% (81% at baseline and 17% after isoproterenol infusion). Parahisian pacing demonstrated that the block was at the AV node level among 150 patients with VA dissociation at baseline. Among patients presenting with supraventricular tachycardia (SVT), 98.7% with atrioventricular nodal reentrant tachycardia (AVNRT) had VA conduction at baseline versus 82.7% presenting with other SVT (atrial fibrillation excluded), P < 0.001. Thus, the absence of VA conduction at baseline during an EPS for SVT carries a negative predictive value (NPV) of 96.9% for AVNRT. VA conduction is present in most patients (98%) with intact AV conduction. The level of VA dissociation when present at baseline is always at the level of the AV node. Patients with SVT presenting for EPS that lacked VA conduction at baseline were unlikely to have AVNRT.


Revising the Diagnosis of Idiopathic Uveitis by Peripheral Blood Transcriptomics.
To test the hypothesis that idiopathic uveitis can be categorized into subtypes based on gene expression from blood. Case control study. We applied RNA-Seq to peripheral blood from patients with uveitis associated with 1 of 4 systemic diseases, including axial spondyloarthritis (n = 17), sarcoidosis (n = 13), inflammatory bowel disease (n = 12), tubulo-interstitial nephritis with uveitis (n = 10), or idiopathic uveitis (n = 38) as well as 18 healthy control subjects evaluated predominantly at Oregon Health and Science University. A high-dimensional negative binomial regression model implemented in the edgeR R package compared each disease group with the control subjects. The 20 most distinctive genes for each diagnosis were extracted. Of 80 genes, there were 75 unique genes. A classification algorithm was developed by fitting a gradient boosting tree with 5-fold cross-validation. Messenger RNA from subjects with idiopathic uveitis were analyzed to see if any fit clinically and by gene expression pattern with one of the diagnosable entities. For uveitis associated with a diagnosable systemic disease, gene expression profiling achieved an overall accuracy of 85% (balanced average of sensitivity plus specificity, P < .001). Although most patients with idiopathic uveitis presumably have none of these 4 associated systemic diseases, gene expression profiles helped to reclassify 11 of 38 subjects. Peripheral blood gene expression profiling is a potential adjunct in accurate differential diagnosis of the cause of uveitis. Validation of these results and characterization of the gene expression profile from additional discrete diagnoses could enhance the value of these observations.


The diversification of terpene emissions in Mediterranean oaks: lessons from a study of Quercus suber, Quercus canariensis and its hybrid Quercus afares.
Interspecific gene flow is common in oaks. In the Mediterranean, this process produced geographical differentiations and new species, which may have contributed to the diversification of the production of volatile terpenes in the oak species of this region. The endemic North African deciduous oak Quercus afares (Pomel) is considered to be a stabilized hybrid between the evergreen Quercus suber (L.) and the deciduous Quercus canariensis (Willd.), presumably being monoterpene and isoprene emitters, respectively. In a common garden experiment, we examined the terpene emission capacities, terpene synthase (TPS) activities and nuclear genetic markers in 52 trees of these three oak species. All but one of the Q. suber and Q. canariensis trees were found to be genetically pure, whereas most Q. afares trees possessed a mixed genotype with a predominance of Q. suber alleles. Analysis of the foliar terpene emissions and TPS activities revealed that all the Q. canariensis trees strongly produced isoprene while all the Q. suber trees were strong monoterpene producers. Quercus afares trees produced monoterpenes as well but at more variable and significantly lower rates, and with a monoterpene pattern different than that observed in Q. suber. Among 17 individuals tested, one Q. afares tree emitted only an insignificant amount of terpenes. No mixed isoprene/monoterpene emitter was detected. Our results suggest that the capacity and pattern of volatile terpene production in Algerian Q. afares populations have strongly diverged from those of its parental species and became quantitatively and qualitatively reduced, including the complete suppression of isoprene production.


Physician Order Entry Clerical Support Improves Physician Satisfaction and Productivity.
To examine the impact of clerical support personnel for physician order entry on physician satisfaction, productivity, timeliness with electronic health record (EHR) documentation, and physician attitudes. All seven part-time physicians at an academic general internal medicine practice were included in this quasi-experimental (single group, pre- and postintervention) mixed-methods study. One full-time clerical support staff member was trained and hired to enter physician orders in the EHR and conduct previsit planning. Physician satisfaction, productivity, timeliness with EHR documentation, and physician attitudes toward the intervention were measured. Four months after the intervention, physicians reported improvements in overall quality of life (good quality, 71%-100%), personal balance (43%-71%), and burnout (weekly, 43%-14%; callousness, 14%-0%). Matched for quarter, productivity increased: work relative value unit (wRVU) per session increased by 20.5% (before, April-June 2014; after, April-June 2015; range -9.2% to 27.5%). Physicians reported feeling more supported, more focused on patient care, and less stressed and fatigued after the intervention. This study supports the use of physician order entry clerical personnel as a simple, cost-effective intervention to improve the work lives of primary care physicians.


Fixed budgets as a cost containment measure for pharmaceuticals.
In Västerbotten County, Sweden, there are two health centers which (in contrast to all other health centers in the region) bear strict responsibility over their pharmaceutical budget. This study examined whether the prices and quantities of pharmaceuticals prescribed by physicians working at these health centers differ significantly from those prescribed by physicians at health centers with open-ended budgets. Estimation results using matching methods, which allows us to compare similar patients at the different health centers, show that the introduction of fixed pharmaceutical budgets did not affect physicians' prescription behavior, indicating that fixed budgets may not be an efficient measure to reduce costs. Another explanation is that the health centers under study already had taken measures to contain costs, making it hard to further reduce costs.


Complex choristoma of the gyrus rectus: a distinct clinicopathologic entity?
We report 2 cases of an unusual central nervous system lesion involving the gyrus rectus and characterized by well-differentiated epithelial and mesenchymal components. One patient was a 30-year-old woman, and the other was a 42-year-old man. Both presented with seizures. Radiologically, the lesions were solid, with ill-defined margins and increased signal on T2-weighted MRI. Both were contrast enhancing. There was no significant mass effect or edema. Histopathologically, the epithelial component was characterized by a tubule-forming, stratified epithelium. The lumina of the tubules were lined, in some areas, by a distinct layer of mature keratinocytes. Smooth muscle actin-immunoreactive myoepithelial cells were present in the basal layer, indicating glandular differentiation. Some lumina were cystically dilated and contained anucleate squames and/or PAS-positive material. In some areas, the epithelial structures were separated by reactive glial tissue resembling pilocytic astrocytoma, with prominent, piloid processes, Rosenthal fibers, and gemistocytic astrocytes. In other areas, glial nuclei formed small clusters reminiscent of subependymoma, but no glioma was identified in either case. Tissue resembling mature adipose was present focally. No mitotic figures were identified. Follow-up interval of up to 2 years showed no evidence of recurrent tumor. By location and histologic features, this lesion appears to represent a rare, distinct clinicopathologic entity, perhaps derived from misplaced cranial neural crest cells.


Genetic Interactions Between Glycine max and Sclerotinia sclerotiorumUsing a Straw Inoculation Method.
Genetic interactions for disease response between cultivars of Glycine max and isolates of Scle-rotinia sclerotiorum were evaluated in controlled-environment inoculations of five soybean cultivars with four genetically unique isolates of S. sclerotiorum. The objective of this study was to identify host-pathogen interactions using isolates of the pathogen which had different geographical and crop-wise distribution as well as a different DNA-based fingerprint. To do so, 4-week-old soybean plants were inoculated with individual isolates of S. sclerotiorum using a straw inoculation method. Inoculated plants were incubated for 48 h in continuous leaf wetness and rated for disease severity 1 and 2 weeks after inoculation. Significant differences in disease severity were detected among the soybean cultivars, and NK S08-80 consistently had the lowest disease severity among the five cultivars tested. No significant differences in disease severity were observed among pathogen isolates and no significant interactions were detected between soybean cultivars and pathogen isolates. These results suggest the following interpretations: (i) either the clonal genotype of the four pathogen isolates as determined by mycelial compatibility and DNA fingerprint was not associated with level of virulence on the five soybean cultivars or (ii) the soybean cultivars themselves were not capable of revealing any differences in virulence among isolates that would be related to their genetic fingerprint or regional distribution. The results of this study are consistent with the practice of considering different isolates of S. scle-rotiorum sampled from soybean in the same geographical region as equivalent for the evaluation of soybean cultivars for resistance to Sclerotinia stem rot.


Structure of a glycerol-conducting channel and the basis for its selectivity.
Membrane channel proteins of the aquaporin family are highly selective for permeation of specific small molecules, with absolute exclusion of ions and charged solutes and without dissipation of the electrochemical potential across the cell membrane. We report the crystal structure of the Escherichia coli glycerol facilitator (GlpF) with its primary permeant substrate glycerol at 2.2 angstrom resolution. Glycerol molecules line up in an amphipathic channel in single file. In the narrow selectivity filter of the channel the glycerol alkyl backbone is wedged against a hydrophobic corner, and successive hydroxyl groups form hydrogen bonds with a pair of acceptor, and donor atoms. Two conserved aspartic acid-proline-alanine motifs form a key interface between two gene-duplicated segments that each encode three-and-one-half membrane-spanning helices around the channel. This structure elucidates the mechanism of selective permeability for linear carbohydrates and suggests how ions and water are excluded.


Geometric control of the cell cycle.
How do cells sense their own size and shape? And how does this information regulate progression of the cell cycle? Our group, in parallel to that of Paul Nurse, have recently demonstrated that fission yeast cells use a novel geometry-sensing mechanism to couple cell length perception with entry into mitosis. These rod-shaped cells measure their own length by using a medially-placed sensor, Cdr2, that reads a protein gradient emanating from cell tips, Pom1, to control entry into mitosis. Budding yeast cells use a similar molecular sensor to delay entry into mitosis in response to defects in bud morphogenesis. Metazoan cells also modulate cell proliferation in response to their own shape by sensing tension. Here I discuss the recent results obtained for the fission yeast system and compare them to the strategies used by these other organisms to perceive their own morphology.


Individual differences in alcohol drinking frequency are associated with electrophysiological responses to unexpected nonrewards.
It has been suggested that alcohol use is related to sensitivity of the reward system. Although there are several studies using self-reported measures supportive of this notion, objective biological data in humans on this issue are lacking. This study is designed to test whether alcohol drinking frequency is associated with electrophysiological indices of reward processing. In a passive gambling task, stimuli predicted the presence (reward) and absence (nonreward) of rewards resulting in P2 and medial frontal negativity (MFN) indices of reward processing. Forty-seven undergraduate students were asked about their habitual drinking frequency and the P2 and MFN to stimuli predicting reward were measured. Most importantly, the MFN to unpredicted nonrewards at the frontal midline (Fz) location correlated significantly with drinking frequency, with frequent drinkers showing larger MFN amplitudes. The results did not show a significant association between frequency and alcohol drinking and P2. Although several studies showing increased reward-sensitivity in addictive behaviors, the present results indicate that, in frequent alcohol drinkers, electrophysiological responsiveness is particularly activated by unpredicted nonrewards. In general, this may point to the involvement of the reward system in alcohol drinking frequency. More specifically, the results demonstrate an increased vulnerability of high frequency drinkers to signals of (frustrative) nonrewards.


Evolutionary Origin of OwlRep, a Megasatellite DNA Associated with Adaptation of Owl Monkeys to Nocturnal Lifestyle.
Rod cells of many nocturnal mammals have a "non-standard" nuclear architecture, which is called the inverted nuclear architecture. Heterochromatin localizes to the central region of the nucleus. This leads to an efficient light transmission to the outer segments of photoreceptors. Rod cells of diurnal mammals have the conventional nuclear architecture. Owl monkeys (genus Aotus) are the only taxon of simian primates that has a nocturnal or cathemeral lifestyle, and this adaptation is widely thought to be secondary. Their rod cells were shown to exhibit an intermediate chromatin distribution: a spherical heterochromatin block was found in the central region of the nucleus although it was less complete than that of typical nocturnal mammals. We recently demonstrated that the primary DNA component of this heterochromatin block was OwlRep, a megasatellite DNA consisting of 187-bp-long repeat units. However, the origin of OwlRep was not known. Here we show that OwlRep was derived from HSAT6, a simple repeat sequence found in the centromere regions of human chromosomes. HSAT6 occurs widely in primates, suggesting that it was already present in the last common ancestor of extant primates. Notably, Strepsirrhini and Tarsiformes apparently carry a single HSAT6 copy, whereas many species of Simiiformes contain multiple copies. Comparison of nucleotide sequences of these copies revealed the entire process of the OwlRep formation. HSAT6, with or without flanking sequences, was segmentally duplicated in New World monkeys. Then, in the owl monkey linage after its divergence from other New World monkeys, a copy of HSAT6 was tandemly amplified, eventually forming a megasatellite DNA.


Combined iron oxide nanoparticle ferumoxytol and gadolinium contrast enhanced MRI define glioblastoma pseudoprogression.
Noninvasively differentiating therapy-induced pseudoprogression from recurrent disease in patients with glioblastoma is prospectively difficult due to the current lack of a biologically specific imaging metric. Ferumoxytol iron oxide nanoparticle MRI contrast characterizes innate immunity mediated neuroinflammation; therefore, we hypothesized that combined ferumoxytol and gadolinium enhanced MRI could serve as a biomarker of glioblastoma pseudoprogression. In this institutional review board-approved, retrospective study, we analyzed ferumoxytol and gadolinium contrast enhanced T1-weighted 3T MRI in 45 patients with glioblastoma over multiple clinical timepoints. Isocitrate dehydrogenase 1 (IDH-1) mutational status was characterized by exome sequencing. Sum of products diameter measurements were calculated according to Response Assessment in Neuro-Oncology criteria from both gadolinium and ferumoxytol enhanced sequences. Enhancement mismatch was calculated as the natural log of the ferumoxytol to gadolinium sum of products diameter ratio. Analysis of variance and Student's t-test assessed differences in mismatch ratios. P-value <0.05 indicated statistical significance. With the development of pseudoprogression we observed a significantly elevated mismatch ratio compared with disease recurrence (P < 0.01) within IDH-1 wild type patients. Patients with IDH-1 mutation demonstrated significantly reduced mismatch ratio with the development of pseudoprogression compared with disease recurrence (P < 0.01). Receiver operator curve analysis demonstrated 100% sensitivity and specificity for the use of mismatch ratios as a diagnostic biomarker of pseudoprogression. Our study suggests that ferumoxytol to gadolinium contrast mismatch ratios are an MRI biomarker for the diagnosis of pseudoprogression in patients with glioblastoma. This may be due to the unique characterization of therapy-induced neuroinflammation.


Exercise intervention to prevent falls and enhance mobility in community dwellers after stroke: a protocol for a randomised controlled trial.
Stroke is the most common disabling neurological condition in adults. Falls and poor mobility are major contributors to stroke-related disability. Falls are more frequent and more likely to result in injury among stroke survivors than among the general older population. Currently there is good evidence that exercise can enhance mobility after stroke, yet ongoing exercise programs for general community-based stroke survivors are not routinely available. This randomised controlled trial will investigate whether exercise can reduce fall rates and increase mobility and physical activity levels in stroke survivors. Three hundred and fifty community dwelling stroke survivors will be recruited. Participants will have no medical contradictions to exercise and be cognitively and physically able to complete the assessments and exercise program. After the completion of the pre-test assessment, participants will be randomly allocated to one of two intervention groups. Both intervention groups will participate in weekly group-based exercises and a home program for twelve months. In the lower limb intervention group, individualised programs of weight-bearing balance and strengthening exercises will be prescribed. The upper limb/cognition group will receive exercises aimed at management and improvement of function of the affected upper limb and cognition carried out in the seated position. The primary outcome measures will be falls (measured with 12 month calendars) and mobility. Secondary outcome measures will be risk of falling, physical activity levels, community participation, quality of life, health service utilisation, upper limb function and cognition. This study aims to establish and evaluate community-based sustainable exercise programs for stroke survivors. We will determine the effects of the exercise programs in preventing falls and enhancing mobility among people following stroke. This program, if found to be effective, has the potential to be implemented within existing community services. The protocol for this study is registered with the Australian New Zealand Clinical Trials Registry (ACTRN12606000479505).


Geostatistical model-based estimates of Schistosomiasis prevalence among individuals aged ≤ 20 years in West Africa.
Schistosomiasis is a water-based disease that is believed to affect over 200 million people with an estimated 97% of the infections concentrated in Africa. However, these statistics are largely based on population re-adjusted data originally published by Utroska and colleagues more than 20 years ago. Hence, these estimates are outdated due to large-scale preventive chemotherapy programs, improved sanitation, water resources development and management, among other reasons. For planning, coordination, and evaluation of control activities, it is essential to possess reliable schistosomiasis prevalence maps. We analyzed survey data compiled on a newly established open-access global neglected tropical diseases database (i) to create smooth empirical prevalence maps for Schistosoma mansoni and S. haematobium for individuals aged ≤ 20 years in West Africa, including Cameroon, and (ii) to derive country-specific prevalence estimates. We used Bayesian geostatistical models based on environmental predictors to take into account potential clustering due to common spatially structured exposures. Prediction at unobserved locations was facilitated by joint kriging. Our models revealed that 50.8 million individuals aged ≤ 20 years in West Africa are infected with either S. mansoni, or S. haematobium, or both species concurrently. The country prevalence estimates ranged between 0.5% (The Gambia) and 37.1% (Liberia) for S. mansoni, and between 17.6% (The Gambia) and 51.6% (Sierra Leone) for S. haematobium. We observed that the combined prevalence for both schistosome species is two-fold lower in Gambia than previously reported, while we found an almost two-fold higher estimate for Liberia (58.3%) than reported before (30.0%). Our predictions are likely to overestimate overall country prevalence, since modeling was based on children and adolescents up to the age of 20 years who are at highest risk of infection. We present the first empirical estimates for S. mansoni and S. haematobium prevalence at high spatial resolution throughout West Africa. Our prediction maps allow prioritizing of interventions in a spatially explicit manner, and will be useful for monitoring and evaluation of schistosomiasis control programs.


Pilocytic astrocytoma of the posterior fossa: a follow-up study in 15 patients.
The extent of resection in pilocytic astrocytoma of the posterior fossa remains undefined, as the problem of hydrocephalus has not yet been solved. We retrospectively reviewed the data from 15 patients with a pilocytic astrocytoma of the posterior fossa to evaluate the impact of surgical technique, in terms of resection extent, by serial magnetic resonance imaging (MRI) examinations. In addition, the issue of hydrocephalus was considered and related to the different treatment modalities. Macroscopic, gross, total resection of the tumor was performed in all 15 patients. Follow-up was obtained in 14 patients for a period ranging between 11 and 119 months (median, 41.5 months). The ages of patients ranged from two to 13 years (mean, 7 +/- 3 years). All patients underwent serial MRI examinations in the first month, every six months for the first two years and then yearly. Outcome was good in 12 patients who had no neurologic deficit and fair in two patients who were slightly handicapped but had an independent life. There were four patients with an abnormally persistent enhancement on MRI, with a median follow-up of 30 months. One of these patients had progressively increasing size of the enhancement. Three of them had the same size of enhancement during the follow-up period. The MRI findings showed residual tumors in four patients. One of them had tumor regrowth one year after surgery. There were 11 cases with pilocytic astrocytoma and hydrocephalus. Five patients were treated with tumor removal and external CSF drainage. Six patients underwent tumor removal only, without perioperative cerebrospinal fluid (CSF) drainage. Only one patient had a permanent ventriculoperitoneal shunt. Our study illustrated that the extent of tumor resection of pilocytic astrocytoma can be defined by postoperative serial MRI examinations. Long-term follow-up with MRI seems mandatory in cases with abnormal enhancement. Hydrocephalus is a common finding in patients with a pilocytic astrocytoma. A permanent ventriculoperitoneal shunt is required only in patients with postoperative hydrocephalus.