On optimal treatment regimes selection for mean survival time.
In clinical studies with time-to-event as a primary endpoint, one main interest is to find the best treatment strategy to maximize patients' mean survival time. Due to patient's heterogeneity in response to treatments, great efforts have been devoted to developing optimal treatment regimes by integrating individuals' clinical and genetic information. A main challenge arises in the selection of important variables that can help to build reliable and interpretable optimal treatment regimes as the dimension of predictors may be high. In this paper, we propose a robust loss-based estimation framework that can be easily coupled with shrinkage penalties for both estimation of optimal treatment regimes and variable selection. The asymptotic properties of the proposed estimators are studied. Moreover, a model-free estimator of restricted mean survival time under the derived optimal treatment regime is developed, and its asymptotic property is studied. Simulations are conducted to assess the empirical performance of the proposed method for parameter estimation, variable selection, and optimal treatment decision. An application to an AIDS clinical trial data set is given to illustrate the method.


The Association Between Patient Safety Culture and Accreditation at Primary Care Centers in Kuwait: A Country-Wide Multi-Method Study.
A culture of patient safety is essential for the continual improvement of service and reducing errors. This study aims to examine how the scores of patient safety culture items impact accreditation compliance percentages in primary care settings in Kuwait. A cross-sectional and a retrospective quantitative approaches were used on 5288 employees at 75 primary healthcare centers across Kuwait. Patient safety culture questionnaires were distributed to clinical and administrative staff with at least one year of experience. We also examined the percentages of compliance with accreditation standards by the studied centers. Participant socio-demographic and setting demographic data were studied using univariate (means, standard deviations, frequencies, percentages) and bivariate (Student's t-tests, ANOVA F-tests, Mann-Whitney U-test, Kruskal-Wallis H-test, Spearman correlation) analyses to examine how trends in the patient safety culture items and accreditation compliance differ across the primary healthcare center demographic groups. The relationship between patient safety culture composites and percentages of accreditation compliance was investigated. To predict the factors for accreditation compliance, we used multivariate regression analysis. The responses of 5288 employees (response rate= 76.5%) from 75 centers were included. Our analysis revealed that the demographics of the setting affected the accreditation compliance percentage more than the percentages of patient safety culture positive ratings. Furthermore, the composites "Patient Care Tracking/Follow-up", "Communication about Error", "Organizational Learning" and "Overall Perceptions of Patient Safety & Quality" had statistically significant weak-to-moderate positive correlations (r = 0.272-0.373) with percentage scores for compliance with various accreditation standards. Regression analysis indicated that the patient safety culture items accounted for 35% to 38% of the variability in accreditation compliance scores. The safety practices adopted by a primary healthcare center reflect its culture, and this has an impact on its accreditation performance. Our interpretation of the findings is expected to help healthcare leaders to better understand the relationship between safety culture and performance and to develop realistic reform strategies.


The effect of subscapularis muscle contraction on coaptation of anteroinferior glenohumeral ligament-labrum complex after Bankart repair.
Facilitation of healing is important for the anteroinferior glenohumeral ligament-labrum complex (AIGHL-LC) after Bankart repair in shoulder dislocation. The purpose of this study was to investigate the effect of subscapularis muscle loading on contact area and contact pressure between the subscapularis and AIGHL-LC and between the glenoid bone and the AIGHL-LC following Bankart repair. Twenty-two fresh-frozen cadaveric shoulders were used. They were attached to a shoulder-positioning device to which a compression force was applied. Loads applied to the supraspinatus, infraspinatus, and teres minor tendons were held constant. The loads applied to the subscapularis tendon were set at 0, 10, 20, and 30 Newton (N). Contact pressure and area between the subscapularis and the AIGHL-LC were measured with the arm at 4 rotational positions: 60° and 30° internal, neutral, and 30° external. After the Bankart lesion was created, the contact area and pressure between the AIGHL-LC and glenoid bone were measured while Bankart repair was performed with or without loading of the subscapularis. The contact area and pressures with 10, 20, and 30 N of subscapularis loadings were significantly greater than with 0 N of subscapularis loading at 60° internal rotation and 30° external rotation (P < .05). After Bankart repair, contact area and pressure with subscapularis loading between the AIGHL-LC and glenoid bone were significantly greater than without subscapularis loading (P < .01). We conclude that isometric contraction exercises of the subscapularis might facilitate healing of the AIGHL-LC after Bankart repair.


Percutaneous Instrumentation of a Complex Lumbar Spine Fracture with Bilateral Pedicle Dissociation: Case Report and Technical Note.
Complex traumatic lumbar spine fractures are difficult to manage and typically occur in younger patients. Surgical immobilization for unstable fractures is an accepted treatment but can lead to future adjacent-level disease. Furthermore, large variations in fracture morphology create significant difficulties when attempting fixation. Therefore, a surgical approach that considers both long-term outcomes and fracture type is of utmost importance. We present a novel technique for percutaneous fixation without interbody or posterolateral fusion in a young patient with bilateral pedicle dissociations and an acute-onset incomplete neurologic deficit. A 20-year-old man involved in a motorcycle accident presented with unilateral right lower extremity paresis and sensory loss with intact rectal tone and no saddle anesthesia. Lumbar computed tomography (CT) demonstrated L2 and L3 fractures associated with bilateral pedicle dislocations. Lumbar magnetic resonance imaging showed draping of the conus medullaris/cauda equina anteriorly over the kyphotic deformity at L2 with minimal associated canal stenosis at L2 and L3. He was treated with emergent percutaneous fixation of the fracture segment without interbody or posterolateral fusion. Decompression was not performed because of the negligible amount of canal stenosis and high likelihood of cerebrospinal fluid leakage due to dural tears from the fractures. Surgical fixation of the L2 vertebra was achieved by cannulating the left pedicle with an oversized tap while holding the right pedicle in place with a normal tap and then driving screws into the left and right pedicles, respectively, thus reducing the free-floating fracture segment. At 18 months after surgery, a follow-up CT demonstrated good cortication across the prior pedicle fractures, and the instrumentation was removed without any obvious signs of instability or disruption of the alignment at the thoracolumbar junction. We present a novel technique for percutaneous reduction and fixation of bilateral pedicle fractures with significant dissociation from the vertebral body, associated neural compression from the kyphotic deformity, and minimal spinal canal stenosis. Furthermore, we argue that early fixation and reduction of the fracture prevented irreversible neurologic compromise, and the absence of interbody or posterolateral fusion ultimately preserved the spinal mobility of the patient once the hardware was removed.


The Wnt/beta-catenin signaling pathway as a target in drug discovery.
The cell signaling cascades provoked by Wnt proteins (the Wnt signaling pathways), which are well conserved through evolution, play crucial roles to maintain homeostasis of a variety of tissues such as skin, blood, intestine, and brain, as well as to regulate proliferation, morphology, motility, and fate of cells during embryonic development. Among these pathways, the signal transduction through beta-catenin (the Wnt/beta-catenin signaling pathway) has been most intensively studied because this signal regulates the expression of a number of genes essential for cell proliferation and differentiation and also this pathway is perturbed in a number of diseases such as cancers, bone diseases, and cardiovascular diseases. However, there is no therapeutic agents that can selectively modulate the Wnt/beta-catenin signaling pathway, although some existing drugs (e.g., non-steroidal anti-inflammatory drugs, vitamins, and imatinib mesylate) have been suggested to inhibit this pathway. Here we provide an overview of the Wnt/beta-catenin signaling pathway: its roles in physiology and pathology and the possibility as a target in development of new drugs.


[Validation of therapeutic test in the diagnosis and treatment of Kala-Azar in children].
The author studied the value of therapeutic test for the diagnosis and treatment of kala-azar in children by comparing two groups of patients. The identification of Leishmania donovani in smears of bone marrow confirmed the diagnosis in all fifty seven patients of the control group. In another group with sixteen children, the diagnosis was based on the successful response to the treatment with meglumine antimonate. In none of the patients of the second group the protozoa was identified and the decision to give the specific treatment was based on epidemiological, clinical, biochemical and hematological criteria for the disease. Statistical analysis was performed by Student t test to compare clinical, biochemical and hematological variables between the groups. None of the differences between the groups was statistically significant, suggesting that the therapeutic test was valid, and all patients who received treatment had kala-azar.


Primary sequence of Xenopus laevis Na+-K+-ATPase and its localization in A6 kidney cells.
Polyclonal antibodies raised against the alpha- and beta-subunits of amphibian kidney Na+-K+-ATPase were used to screen an expression library from Xenopus laevis kidney epithelial cells (A6 cell line). cDNAs coding for each Na+-K+-ATPase subunit were identified and used to isolate near full-length cDNAs. The complete nucleotide sequence and the deduced amino acid sequence were determined. The alpha-subunit is an alpha (alpha I)-isoform. The alpha- and beta-subunits are more closely related to the mammalian and avian than the fish sequences. Antibodies raised against the fusion proteins produced by the two clones served to immunoprecipitate proteins from biosynthetically labeled or selectively surface-radioiodinated A6 cells grown on a porous substrate. The alpha- and the beta-subunits of Na+-K+-ATPase were found associated early in the course of biosynthesis and were restricted to the basolateral plasma membrane.


Transcriptomic Profiling of Duodenal Epithelium Reveals Temporally Dynamic Impacts of Direct Duodenal Starch-Infusion During Dry Period of Dairy Cattle.
Previous research has demonstrated a positive relationship between dietary Metabolisable Energy Intake (MEI) and increased maintenance energy costs associated with the visceral tissues. Limitations in understanding this relationship include a lack of access to samples to assess regulatory control of the putative response gastrointestinal tissues to nutrients. This experiment was conducted with a single nutrient (starch hydrolysate) infused (7 d) directly into the intestine to mimic typical changes in post-ruminal starch delivery in dairy production settings. Duodenal epithelial samples collected via biopsy were evaluated using next-generation sequencing technology (RNA-Seq) to validate the use of this approach for the profiling and comparison of the transcriptome of cattle intestinal epithelial tissues. Samples of intestinal epithelial tissue were collected prior to and during the infusion of starch hydrolysate. Biopsies were collected on day 0 before and day 1, day 3, and day 7 during the infusion. Additionally, samples were collected on day 1 and day 7 after infusion was discontinued (Day 8 and Day14 of the experiment). Evaluation of RNA-seq data revealed dynamic changes in global gene expression during infusion. On day 7 of the infusion, 1490 genes were found to be differentially expressed (DE) compared to the day 0 control samples with FDR p < 0.05, vs. 105 genes on day 1 and 246 genes on Day 3. However, on day 8, after infusion was terminated for 24 h, only 428 genes were identified as differentially expressed compared to day 0 and only 107 genes continued to be identified by Day 14. Thus, the apparent differential expression of these genes is putatively a result of the single nutrient infused. Further, performing function and pathway analysis of the identified DE genes using IPA, we observe changes in digestive system development, and function pathways are among the primary functions of the DE genes, as well as immune response elements. Finally, primary transcription regulators such as PTH, JUN, WNT, and TNFRSF11B were identified as the activated upstream regulators for specific future focus. Using a serial biopsy approach we are able to identify differentially expressed genes from cow duodenal epithelial tissue in response to a short-term perturbation with infused starch hydrolysate.


Intranasal and intramuscular proteosome-staphylococcal enterotoxin B (SEB) toxoid vaccines: immunogenicity and efficacy against lethal SEB intoxication in mice.
Intranasal or intramuscular (i.m.) immunization of mice and i.m. immunization of rabbits with formalinized staphylococcal enterotoxin B (SEB) toxoid in saline elicited higher anti-SEB serum immunoglobulin G (IgG) titers when the toxoid was formulated with proteosomes. In addition, intranasal immunization of mice with this proteosome-toxoid vaccine elicited high levels of anti-SEB IgA in lung and intestinal secretions, whereas the toxoid without proteosomes did not. Two i.m. immunizations with proteosome-toxoid plus alum also induced higher murine serum responses than alum-adjuvanted toxoid without proteosomes. Furthermore, proteosome-toxoid delivered intranasally in saline or i.m. with either saline or alum afforded significant protection against lethal SEB challenge in two D-galactosamine-sensitized murine models of SEB intoxication, i.e., the previously described i.m. challenge model and a new respiratory challenge model of mucosal SEB exposure. Efficacy correlated with the induction of high serum levels of anti-SEB IgG. In contrast, intranasal or i.m. immunization with toxoid in saline without proteosomes was not significantly protective in either challenge model. Proteosome-toxoid plus alum given i.m. also elicited more significant protection against respiratory challenge than the alum-adjuvanted toxoid alone. The capacity of proteosomes to enhance both i.m. and intranasal immunogenicity and efficacy of SEB toxoid indicates that testing such proteosome-SEB toxoid vaccines in the nonhuman primate aerosol challenge model of SEB intoxication prior to immunogenicity trials in humans is warranted. These data expand the applicability of the proteosome mucosal vaccine delivery system to protein toxoids and suggest that respiratory delivery of proteosome vaccines may be practical for enhancement of both mucosal and systemic immunity against toxic or infectious diseases.


[Cerebral actinomycosis. Anatomo-clinical case report and review of the Brazilian literature].
A case of central nervous system actinomycosis is reported. A 33-year-old male complained of headache, vomiting and blurred vision lasting for eight days prior to admission. On examination, a right hemiparesis, as well an intracranial hypertension were detected. The cerebrospinal fluid showed mild lymphomononuclear hypercytosis. Necropsy disclosed three abscess in the cerebral hemispheres, in addition to moderate cerebral edema on the left side but without purulent leptomeningitis. Actinomyces filaments and granules were demonstrated in the cerebral and lung abscessess. The Brazilian literature on actinomycosis is reviewed and six published cases with nervous system involvement were found. Relevant clinical and anatomical aspects of the cases and of the present one are discussed.


Primary aldosteronism: functional histopathology and long-term follow-up after unilateral adrenalectomy.
To investigate the long-term outcome after unilateral adrenalectomy in patients with primary aldosteronism (PA) and to establish the role of functional pathology for the final diagnosis of aldosterone-producing adenoma (APA) or hyperplasia. A single-centre, retrospective cohort study in a hospital setting. Consecutive patients with PA, n = 120, who underwent unilateral adrenalectomy between 1985 and 2010. Preoperative and postoperative data were analysed. To indicate the site of aldosterone secretion in the resected adrenal, we added functional methods to routine histopathology, using in situ hybridization and immunohistochemistry to detect the presence of enzymes needed for aldosterone (CYP11B2) and cortisol (CYP11B1, CYP17A1) synthesis. The median follow-up was 5 years and the cure rate of PA 91%. Hypertension was improved in 97% and normalized in 38%. Functional histopathology changed the final diagnosis from APA to hyperplasia in 6 cases (7%). Five of these had no expression of or staining for aldosterone synthase in the adenoma, but only in nodules in the adjacent cortex. All except one APA patient were cured of PA. They had lower preoperative serum potassium and higher 24-h urinary aldosterone than patients with hyperplasia. Among patients with hyperplasia 16 of 26 were cured. Most patients were cured of PA by unilateral adrenalectomy. Almost all noncured benefitted from the operation as the blood pressure improved. Functional histopathology proved helpful in the distinction between APA and hyperplasia, and we recommend that functional histopathology should be added to routine histopathology to improve the diagnostic evaluation and aid in tailoring the follow-up.


[The evaluation of prognostic value of biomarkers in patients with chronic cardiac insufficiency].
Nowadays, the laboratory markers of myocardial dysfunction (cerebral natriuretic peptide - NT-proBNP); instability of atherosclerotic plaque (highly sensitive C-reactive protein - hsCRP); damages of cardiac muscle (highly sensitive cardiac tropine I - hs-cTnl) play a key role in diagnostic, course prognostics and verification of risk of unfavorable outcomes in patients with chronic cardiac insufficiency. The article presents the results of study of dynamics of levels of NT-proBNP, hsCRP, hs-cTnl in 71 patients with chronic cardiac insufficiency of II and III functional class (according classification of New-York association of cardiologists - NYHA). The comparison was made concerning analyzed laboratory markers with fraction of output of left ventricle of heart and index of body mass relatively to their prognostic role inpatients with chronic cardiac insufficiency with different clinical conditions and various outcomes of disease. It is demonstrated that level of hs-cTnl is the most valuable in respect to unfavorable prognosis of course of chronic cardiac insufficiency and risk of lethal outcome. The level of NT-proBNP has invert correlation relationship with fraction of output of left ventricle of heart and can be considered as a laboratory indicator of functional condition of myocardium in patients with chronic cardiac insufficiency.


Adult congenital heart disease: experience with the surgical approach.
To report the institution experience with the surgical treatment of adults with congenital heart disease due to the increasing number of these patients and the need for a better discussion of the subject. Retrospective analysis describing demographic data, risk factors and results. 191 patients between 16 and 74 years old were operated on. Primary correction was done in 171 cases, 93 (55%) for atrial septal defect repair. Among 20 (12%) reoperations, pulmonary valve replacement was done in six cases. The mean intensive care and hospital stay were 2.7 and 8.5 days respectively, significantly greater for the reoperated cases (P=0.001). The mean bypass and clamping times were 68.6 and 44.7 minutes respectively, greater for the reoperated cases (P<0.0001 and P=0.0003 respectively). Hospital mortality was 4.2% and male sex, functional class III-IV and older age at operation were predictive risk factors. Significant complications were more frequent in the reoperated cases (P<0.003), mainly atrial flutter and fibrillation. Among 183 patients discharged, 149 (82%) are being followed and atrial flutter and fibrillation are common. The mean functional class value improved significantly after operation (1.66 to 1.11; P<0.0001). The estimated survival was 96.2% in six years. Heart surgery in adults with congenital heart disease can be accomplished with low mortality and functional class improvement. Immediate and late complications are frequent. Multicenter studies are important to better characterize this patient population in the country.


Top-down effects of a terrestrial frog on forest nutrient dynamics.
Many studies have found top-down effects of predators on prey, but few studies have linked top-down effects of vertebrate predators to nutrient cycling rates in terrestrial systems. In this study, large and significant effects of a terrestrial frog, Eleutherodactylus coqui (coquí), were recorded on nutrient concentrations and fluxes in a subtropical wet forest. In a manipulative experiment, coquís at natural densities were contained in or excluded from 1 m3 enclosures for 4 months. Chemistry of leaf wash (throughfall), foliage, and decomposed leaf litter in the enclosures were measured as indicators of coquí effects on nutrient cycling. Coquí exclusion decreased elemental concentrations in leaf washes by 83% for dissolved organic C, 71% for NH4+, 33% for NO3-, 60% for dissolved organic N, and between 60 and 100% for Ca, Fe, Mg, Mn, P, K, and Zn. Coquí exclusion had no effect on foliar chemistry of plants transplanted into the enclosures. However, coquí exclusion decreased nutrient availability in decomposing mixed leaf litter by 12% and 14% for K and P, respectively, and increased C:N ratios by 13%. Changes in nutrient concentrations that occurred with coquí exclusion appear to be due to concentrations of nutrients in coquí waste products and population turnover. The results supported our hypothesis that coquís have an observable effect on nutrient dynamics in this forest. We suggest that the primary mechanism through which they have this effect is through the conversion of insects into nutrient forms that are more readily available for microbes and plants. The potential for higher trophic level species to affect nutrient cycling through this mechanism should not be overlooked.


An experiment on standardized cell culture assay in assessing the activities of Composite Artemisia Capillaris Tablets against hepatitis B virus replication in vitro.
To explore the activities of Composite Artemisia Capillaris Tablet (CACT) against hepatitis B virus replication in vitro. By means of radioimmunoassay (RIA), Dot blot and Southern blot, the surface and e antigen production of 2.2.15 cells, HBV DNA in 2.2.15 cell culture medium and that in 2.2.15 cells were examined respectively. HBsAg, HBeAg values of 2.2.15 cells treated by CACT were lower than those of the control, the HBV DNA quantities in culture medium and in 2.2.15 cells decreased as compared with those cells with no treatment by CACT given to them. CACT could inhibit HBV DNA replication, showing its potential antiviral activity in hepatitis B treatment.


Characteristics of self-regulation of the epidemic process of infection caused by the Epstein-Barr virus (Herpesviridae: Lymphocryptovirus, HHV-4).
Among the available scientific literature, there are no publications addressing processes of self-regulation in the parasite-host population systems with reference to chronic infections, including the infection caused by the Epstein-Barr virus (EBV infection). The aim of the study is to assess manifestations of the epidemic process of chronic EBV infection through the lens of the basic tenets of the theory of self-regulation of parasitic systems. The study was performed using data from scientific publications selected from such database sources as Scopus, Web of Science, Cochrane Library, PubMed, CyberLeninka, RSCI, etc. The list of analyzed publications included published articles of the authors of this study, reporting the results of the retrospective epidemiological analysis of the incidence of infectious mononucleosis in Russia in general and in Moscow in particular, as well as the results of the laboratory tests regarding the detection frequency of specific antibodies to EBV proteins. The chronic course of EBV infection promotes a close long-term interaction between the pathogen and the host. The genetic variability of the pathogen and the functions of specific and nonspecific human immune defense systems play a key role in the interaction between two heterogeneous populations and underlie their phasal self-transformation. A variety of social and natural factors (adverse chemical, physical, biological, climatic impacts, etc.) trigger the reactivation of chronic EBV infection, thus providing the continuous existence of additional sources of infection in the host population. The analysis of the manifestations of chronic EBV infection in the context of the theory of self-regulation of parasitic systems promotes the understanding of the factors underlying the unevenness of its epidemic process. The obtained data can be adjusted for other infections having similar transmission mechanisms and virus life cycles (including other herpes infections) to map out strategies to control the epidemic process of chronic infections spread by aerosol transmission of the pathogen.


The effect of fresh gas flow and anesthetic technique on the ability to control acute hemodynamic responses during surgery.
We evaluated the effect of the fresh gas flow (FGF) rate and the anesthetic technique on the ability to control the acute hyperdynamic response to a specific surgical stimulus during surgery in 90 consenting ASA physical status I-III patients undergoing lower abdominal procedures. After the administration of midazolam 2 mg IV, anesthesia was induced in all patients with propofol 1.5 mg/kg IV and fentanyl 1 microg/kg IV and was initially maintained with desflurane or isoflurane, 0.7 minimum alveolar anesthetic concentration, at total FGF rates of either 1 or 3 L/min. In response to the surgical stimulation of skin incision and retropubic dissection, an increase in mean arterial pressure (MAP) >20% above the preincision baseline MAP value provoked a stepwise increase in the inspired concentration of the volatile anesthetic or the IV administration of a variable-rate infusion of esmolol. At both FGF rates, the acute hemodynamic response to surgical stimulation was more efficiently treated by increasing the inspired concentration of desflurane than isoflurane. At 1 L/min, the average time to control the increase in MAP was significantly shorter with desflurane (17+/-12 min) compared with isoflurane (29+/-16 min), with 60% of the patients in the isoflurane group requiring rescue therapy. When an esmolol infusion was used to control the increase in MAP, supplementation with fentanyl was required in 40% and 53% of patients anesthetized with desflurane and isoflurane, respectively. In conclusion, desflurane provided more rapid and reliable control of acute hemodynamic responses to surgical stimulation than isoflurane or esmolol when the volatile anesthetics were administered at low FGF rates. At low fresh gas flow rates (1 L/min), desflurane more successfully and rapidly controlled the acute hemodynamic responses to painful surgical stimuli than isoflurane.


Estimating neural activity from visual areas using functionally defined EEG templates.
Electroencephalography (EEG) is a common and inexpensive method to record neural activity in humans. However, it lacks spatial resolution making it difficult to determine which areas of the brain are responsible for the observed EEG response. Here we present a new easy-to-use method that relies on EEG topographical templates. Using MRI and fMRI scans of 50 participants, we simulated how the activity in each visual area appears on the scalp and averaged this signal to produce functionally defined EEG templates. Once created, these templates can be used to estimate how much each visual area contributes to the observed EEG activity. We tested this method on extensive simulations and on real data. The proposed procedure is as good as bespoke individual source localization methods, robust to a wide range of factors, and has several strengths. First, because it does not rely on individual brain scans, it is inexpensive and can be used on any EEG data set, past or present. Second, the results are readily interpretable in terms of functional brain regions and can be compared across neuroimaging techniques. Finally, this method is easy to understand, simple to use and expandable to other brain sources.


Linking crop traits to transcriptome differences in a progeny population of tetraploid potato.
Potato is the third most consumed crop in the world. Breeding for traits such as yield, product quality and pathogen resistance are main priorities. Identifying molecular signatures of these and other important traits is important in future breeding efforts. In this study, a progeny population from a cross between a breeding line, SW93-1015, and a cultivar, Désirée, was studied by trait analysis and RNA-seq in order to develop understanding of segregating traits at the molecular level and identify transcripts with expressional correlation to these traits. Transcript markers with predictive value for field performance applicable under controlled environments would be of great value for plant breeding. A total of 34 progeny lines from SW93-1015 and Désirée were phenotyped for 17 different traits in a field in Nordic climate conditions and controlled climate settings. A master transcriptome was constructed with all 34 progeny lines and the parents through a de novo assembly of RNA-seq reads. Gene expression data obtained in a controlled environment from the 34 lines was correlated to traits by different similarity indices, including Pearson and Spearman, as well as DUO, which calculates the co-occurrence between high and low values for gene expression and trait. Our study linked transcripts to traits such as yield, growth rate, high laying tubers, late and tuber blight, tuber greening and early flowering. We found several transcripts associated to late blight resistance and transcripts encoding receptors were associated to Dickeya solani susceptibility. Transcript levels of a UBX-domain protein was negatively associated to yield and a GLABRA2 expression modulator was negatively associated to growth rate. In our study, we identify 100's of transcripts, putatively linked based on expression with 17 traits of potato, representing both well-known and novel associations. This approach can be used to link the transcriptome to traits. We explore the possibility of associating the level of transcript expression from controlled, optimal environments to traits in a progeny population with different methods introducing the application of DUO for the first time on transcriptome data. We verify the expression pattern for five of the putative transcript markers in another progeny population.


A novel probe for the non-invasive detection of tumor-associated inflammation.
A novel dual-mode contrast agent was formulated through the addition of an optical near infrared (NIR) probe to a perfluorocarbon (PFC)-based 19F magnetic resonance imaging (MRI) agent, which labels inflammatory cells in situ. A single PFC-NIR imaging agent enables both a qualitative, rapid optical monitoring of an inflammatory state and a quantitative, detailed and tissue-depth independent magnetic resonance imaging (MRI). The feasibility of in vivo optical imaging of the inflammatory response was demonstrated in a subcutaneous murine breast carcinoma model. Ex vivo optical imaging was used to quantify the PFC-NIR signal in the tumor and organs, and results correlated well with quantitative 19F NMR analyses of intact tissues. 19F MRI was employed to construct a three-dimensional image of the cellular microenvironment at the tumor site. Flow cytometry of isolated tumor cells was used to identify the cellular localization of the PFC-NIR probe within the tumor microenvironment. Contrast is achieved through the labeling of host cells involved in the immune response, but not tumor cells. The major cellular reservoir of the imaging agent were tumor-infiltrating CD11b+ F4/80low Gr-1low cells, a cell subset sharing immunophenotypic features with myeloid-derived suppressor cells (MDSCs). These cells are recruited to sites of inflammation and are implicated in immune evasion and tumor progression. This PFC-NIR contrast agent coupled to non-invasive, quantitative imaging techniques could serve as a valuable tool for evaluating novel anticancer agents.


Development of an ANN optimized mucoadhesive buccal tablet containing flurbiprofen and lidocaine for dental pain.
A novel mucoadhesive buccal tablet containing flurbiprofen (FLB) and lidocaine HCl (LID) was prepared to relieve dental pain. Tablet formulations (F1-F9) were prepared using variable quantities of mucoadhesive agents, hydroxypropyl methyl cellulose (HPMC) and sodium alginate (SA). The formulations were evaluated for their physicochemical properties, mucoadhesive strength and mucoadhesion time, swellability index and in vitro release of active agents. Release of both drugs depended on the relative ratio of HPMC:SA. However, mucoadhesive strength and mucoadhesion time were better in formulations, containing higher proportions of HPMC compared to SA. An artificial neural network (ANN) approach was applied to optimise formulations based on known effective parameters (i.e., mucoadhesive strength, mucoadhesion time and drug release), which proved valuable. This study indicates that an effective buccal tablet formulation of flurbiprofen and lidocaine can be prepared via an optimized ANN approach.


Distinct interactions stabilize EGFR dimers and higher-order oligomers in cell membranes.
The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase (RTK) with important roles in many cellular processes as well as cancer and other diseases. EGF binding promotes EGFR dimerization and autophosphorylation through interactions that are well understood structurally. However, it is not clear how these dimers relate to higher-order EGFR oligomers detected at the cell surface. We used single-particle tracking (SPT) and Förster resonance energy transfer (FRET) imaging to examine how each domain within EGFR contributes to receptor dimerization and the rate of its diffusion in the cell membrane. We show that the EGFR extracellular region is sufficient to drive receptor dimerization, but that the EGF-induced EGFR slow-down seen by SPT requires formation of higher order oligomers, mediated in part by the intracellular tyrosine kinase domain - but only when in its active conformation. Our data thus provide important insight into higher-order EGFR interactions required for EGF signaling.


Isolation, culture, and characterization of chicken intestinal epithelial cells.
Enterocytes exert an absorptive and protective function in the intestine, and they encounter many different challenging factors such as feed, bacteria, and parasites. An intestinal epithelial in vitro model can help to understand how enterocytes are affected by these factors and contribute to the development of strategies against pathogens. The present study describes a novel method to culture and maintain primary chicken enterocytes and their characterization by immunofluorescence and biomolecular approaches. Starting from 19-day-old chicken embryos it was possible to isolate viable intestinal cell aggregates that can expand and produce a self-maintaining intestinal epithelial cell population that survives until 12 days in culture. These cells resulted positive in immunofluorescence to Cytokeratin 18, Zonula occludens 1, Villin, and Occludin that are common intestinal epithelial markers, and negative to Vimentin that is expressed by endothelial cells. Cells were cultured also on Transwell® permeable supports and trans-epithelial electrical resistance, was measured. This value gradually increased reaching 64 Ω*cm2 7 days after seeding and it remained stable until day 12. Based on these results it was confirmed that it is possible to isolate and maintain chicken intestinal epithelial cells in culture and that they can be suitable as in vitro intestinal model for further studies.


Imaging inflammation in mouse colon using a rapid stage-scanning confocal fluorescence microscope.
Large area confocal microscopy may provide fast, high-resolution image acquisition for evaluation of tissue in pre-clinical studies with reduced tissue processing in comparison to histology. We present a rapid beam and stage-scanning confocal fluorescence microscope to image cellular and tissue features along the length of the entire excised mouse colon. The beam is scanned at 8,333 lines/sec by a polygon scanning mirror while the specimen is scanned in the orthogonal axis by a motorized translation stage with a maximum speed of 7 mm/sec. A single 1 × 60 mm(2) field of view image spanning the length of the mouse colon is acquired in 10 s. Z-projection images generated from axial image stacks allow high resolution imaging of the surface of non-flat specimens. In contrast to the uniform size, shape, and distribution of colon crypts in confocal images of normal colon, confocal images of chronic bowel inflammation exhibit heterogeneous tissue structure with localized severe crypt distortion.


Efficacy of detergents in removing Salmonella and Shigella spp. from the surface of fresh produce.
Fresh produce has been implicated in several foodborne disease outbreaks. Produce surfaces can be primary sites of contamination during production and handling. One approach to reduce contamination is to treat fresh produce with rinsing agents. In this study, different detergent agents were used at 22 and 40 degrees C to determine their efficacy in removing Salmonella and Shigella spp. from the surfaces of strawberries, tomatoes, and green-leaf lettuce. Produce was inoculated at 22 degrees C with a cocktail of nalidixic acid-resistant organisms (6 to 6.5 log CFU/ml). After air drying for 1 h, samples were rinsed with either 0.1% Tween 80, 0.1% sodium lauryl sulfate (SLS), or water (control) at 22 or 40 degrees C. Rinse solutions were spiral plated onto tryptic soy agar supplemented with 50 mg of nalidixic acid per liter. In trials involving strawberries and lettuce, Salmonella and Shigella were removed at levels of 4 and 3 log CFU/ml, respectively, except from Salmonella-inoculated strawberries rinsed with SLS, for which minimal removal rates were 1.5 log CFU/ml at 22 degrees C and < 1 log CFU/ml at 40 degrees C. When whole strawberries were analyzed after rinsing with SLS, few organisms were recovered. This result suggests that SLS may have a lethal or sublethal effect on Salmonella, especially when a 40 degrees C solution is used. Salmonella and Shigella removal rates for tomatoes were 1 and 1.5 log CFU/ml lower, respectively, than those for strawberries or lettuce. Overall, detergents were no more effective in removing organisms from produce than water was. The detergents examined would not constitute effective overall produce rinse treatments.


Pharmacokinetics of dexmedetomidine during administration of vatinoxan in male neutered cats anesthetized with isoflurane.
Dexmedetomidine is an alpha-2 adrenoceptor agonist, and vatinoxan is an alpha-2 antagonist believed to poorly cross the blood-brain barrier in cats. Dexmedetomidine-vatinoxan combinations are of interest in anesthetized cats because the anesthetic sparing effect of dexmedetomidine may be preserved while vatinoxan attenuates the adverse cardiovascular effects of dexmedetomidine. The aim of this study was to characterize the pharmacokinetics of dexmedetomidine in cats during administration of isoflurane and vatinoxan. Six healthy adult male castrated cats were anesthetized with isoflurane in oxygen. Vatinoxan was administered using a target-controlled infusion system intended to maintain a plasma concentration of 4 µg/ml. Dexmedetomidine, 35 µg/kg was administered intravenously over 5 min. Plasma dexmedetomidine and vatinoxan concentrations were measured at selected time points ranging from prior to 8 hr after dexmedetomidine administration using liquid chromatography/tandem mass spectrometry. Compartment models were fitted to the time-concentration data using nonlinear mixed-effect modeling. A three-compartment model best fitted the data. Typical value (% interindividual variability) for the three-compartment volumes (ml/kg), the metabolic clearance and the two intercompartment distribution clearances (ml min-1 kg-1 ) were 168 (259), 318 (35), 1,425 (18), 12.4 (31), 39.1 (18), and 29.6 (17), respectively. Mean ± standard deviation plasma vatinoxan concentration was 2.6 ± 0.6 µg/ml.


Trimodal gadolinium-gold microcapsules containing pancreatic islet cells restore normoglycemia in diabetic mice and can be tracked by using US, CT, and positive-contrast MR imaging.
To develop microcapsules that immunoprotect pancreatic islet cells for treatment of type I diabetes and enable multimodal cellular imaging of transplanted islet cells. All animal experiments were approved by the institutional animal care and use committee. Gold nanoparticles functionalized with DTDTPA (dithiolated diethylenetriaminepentaacetic acid):gadolinium chelates (GG) were coencapsulated with pancreatic islet cells by using protamine sulfate as a clinical-grade alginate cross linker. Conventional poly-l-lysine-cross-linked microcapsules and unencapsulated islets were included as controls. The viability and glucose responsiveness of islet cells were assessed in vitro, and in vivo insulin (C-peptide) secretion was monitored for 6 weeks in (streptozotocin-induced) diabetic mice with (n = 7) or without (n = 8) intraabdominally engrafted islet cells. Five nondiabetic mice were included as controls. Differences between samples were calculated by using a nonparametric Wilcoxon Mann-Whitney method. To adjust for multiple comparisons, a significance level of P < .01 was chosen. Generalized estimating equations were used to model cell function over time. Three mice with engrafted capsules were imaged in vivo with high-field-strength (9.4-T) magnetic resonance (MR) imaging, micro-computed tomography (CT), and 40-MHz ultrasonography (US). Encapsulated human pancreatic islets were functional in vitro for at least 2 weeks after encapsulation. Blood glucose levels in the diabetic mice transplanted with GG-labeled encapsulated mouse βTC6 insulinoma cells returned to normal within 1 week after transplantation, and normoglycemia was sustained for at least 6 weeks without the use of immunosuppressive drugs. GG microcapsules could be readily visualized with positive-contrast high-field-strength MR imaging, micro-CT, and US both in vitro and in vivo. Cell encapsulation with GG provides a means of trimodal noninvasive tracking of engrafted cells.


Glucose metabolism gene expression patterns and tumor uptake of ¹⁸F-fluorodeoxyglucose after radiation treatment.
To investigate whether radiation treatment influences the expression of glucose metabolism genes and compromises the potential use of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) as a tool to monitor the early response of head and neck cancer xenografts to radiation therapy (RT). Low passage head and neck squamous cancer cells (UT14) were injected to the flanks of female nu/nu mice to generate xenografts. After tumors reached a size of 500 mm(3) they were treated with either sham RT or 15 Gy in 1 fraction. At different time points, days 3, 9, and 16 for controls and days 4, 7, 12, 21, 30, and 40 after irradiation, 2 to 3 mice were assessed with dynamic FDG-PET acquisition over 2 hours. Immediately after the FDG-PET the tumors were harvested for global gene expression analysis and immunohistochemical evaluation of GLUT1 and HK2. Different analytic parameters were used to process the dynamic PET data. Radiation had no effect on key genes involved in FDG uptake and metabolism but did alter other genes in the HIF1α and glucose transport-related pathways. In contrast to the lack of effect on gene expression, changes in the protein expression patterns of the key genes GLUT1/SLC2A1 and HK2 were observed after radiation treatment. The changes in GLUT1 protein expression showed some correlation with dynamic FDG-PET parameters, such as the kinetic index. (18)F-fluorodeoxyglucose positron emission tomography changes after RT would seem to represent an altered metabolic state and not a direct effect on the key genes regulating FDG uptake and metabolism.


Measuring ovarian function in young cancer survivors.
It is known that young female cancer survivors are at higher risk for decreased fertility and early menopause. Because the risk of these outcomes varies by age, cancer type, treatment regimen and other patient-specific characteristics, there is a need for valid tools for measuring and predicting reproductive function in this population. Ovarian reserve tests (ORT) include serum and ultrasound biomarkers used in characterizing healthy ovarian aging and predicting ovarian reserve prior to fertility treatment. This review summarizes efforts to translate ORT to use in female cancer patients. To date, small longitudinal and cross-sectional studies of young cancer patients demonstrate that gonadotoxic chemotherapy impact ORT. While follicle stimulating hormone (FSH), anti-Mullerian hormone (AMH), inhibin B, antral follicle count (AFC) and ovarian volume have all been demonstrated to change with exposure to gonadotoxic therapy, FSH, AMH and AFC appear to be the most sensitive. More data are needed to determine the long-term effect of hormonal agents from tamoxifen to GnRH agonists on ORT. The effect of unilateral oophorectomy acutely or in the long-term on ORT is not known. There are some early data postulating that cancer itself may impair ovarian reserve. Among cancer survivors, ORT correlate with amenorrhea. Even in young survivors with normal menstrual cycles, hormone and ultrasound measures of ovarian reserve suggest decreased underlying ovarian reserve than age-matched healthy women. More studies are needed to determine the optimal time to test ORT in the large number of young survivors on combined estrogen and progesterone contraceptives. A significant utility of ORT would be to be able to predict fertility for young survivors, and there are no data on this outcome to date. The goal of future studies of ORT in young girls and women with cancer is to determine their utility as surrogate measures of ovarian function or predictors of infertility or ovarian failure. To do so, large-scale data need to be collected through cooperative group mechanisms. Epidemiologically, studies need to move from association studies to develop ORT into appropriate screening and predictive tests.


Spiro Rhodamine-Perylene Compact Electron Donor-Acceptor Dyads: Conformation Restriction, Charge Separation, and Spin-Orbit Charge Transfer Intersystem Crossing.
Spiro rhodamine (Rho)-perylene (Pery) electron donor-acceptor dyads were prepared to study the spin-orbit charge transfer intersystem crossing (SOCT-ISC) in these rigid and sterically congested molecular systems. The electron-donor Rho (lactam form) moiety is attached via the N-C bond to the electron acceptor at either 1- or 3-position of the Pery moiety (Rho-Pery-1 and Rho-Pery-3). Severe torsion of the Pery moiety in Rho-Pery-1 was observed. The fluorescence of the two dyads is significantly quenched in polar solvents, and the singlet oxygen quantum yields (ΦΔ) are strongly dependent on solvent polarity (4-36%). Femtosecond transient absorption spectra demonstrate that charge separation (CS) takes 0.51 ps in Rho-Pery-1 and 5.75 ps in Rho-Pery-3, and the charge recombination (CR)-induced ISC is slow (>3 ns). Nanosecond transient absorption spectra indicate that the formation of triplet states via SOCT-ISC takes 24-75 ns for Rho-Pery-1 and 6-15 ns for Rho-Pery-3, and the distorted π-framework of the Pery moiety results in a shorter triplet lifetime of 19.9 vs 291 μs for the planar analogue. Time-resolved electron paramagnetic resonance spectroscopy confirms the SOCT-ISC mechanism.


Meningoencephalitis caused by concurrent infection with canine distemper virus and a unique Sarcocystis sp. in a gray fox.
A deceased 9-wk-old male gray fox (Urocyon cinereoargenteus) with a history of decreased ambulation and diarrhea was submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory. No significant gross findings were evident on postmortem examination. Histologically, the cerebrum and brainstem had mild necrotizing meningoencephalitis with protozoal schizonts and merozoites. Additionally, glial cells contained intracytoplasmic and intranuclear viral inclusion bodies. Sections of the cerebrum were positive for canine distemper virus (CDV) and negative for Sarcocystis neurona on immunohistochemistry. Bayesian analysis revealed that this Sarcocystis sp. clustered most closely with a clade of unnamed Sarcocystis sp. found in viperid snakes, with a posterior probability of 99%. CDV likely played a significant role in the expression of clinical sarcocystosis in this gray fox.


Rodent Ectoparasites in the Middle East: A Systematic Review and Meta-Analysis.
Rodents carry many ectoparasites, such as ticks, lice, fleas, and mites, which have potential public health importance. Middle Eastern countries are hotspots for many emerging and re-emerging infectious diseases, such as plague, leishmaniasis, Crimean Congo hemorrhagic fever, and Q fever, due to their ecological, socioeconomic, and political diversity. Rodent ectoparasites can act as vectors for many of these pathogens. Knowledge of rodent ectoparasites is of prime importance in controlling rodent ectoparasite-borne zoonotic diseases in this region. The current systematic review and meta-analysis performs a comprehensive synthesis of the available knowledge, providing an evidence-based overview of the ectoparasites detected on rodents in Middle Eastern countries. Following a systematic search in Pubmed, Scopus, and Web of Science, a total of 113 published articles on rodent ectoparasites were studied and analyzed. A total of 87 rodent species were documented, from which Mus musculus, Rattus norvegicus, and Rattus rattus were found to be the most common. Fleas were the most reported ectoparasites (87 articles), followed by mites (53), ticks (44), and lice (25). Xenopsylla cheopis, Polyplax spinulosa, Ornithonyssus bacoti, and Hyalomma rhipicephaloides were the most commonly described fleas, lice, mites, and ticks, respectively. Based on the reviewed articles, the median flea, louse, mite, and tick indices were highest in Israel (4.15), Egypt (1.39), Egypt (1.27), and Saudi Arabia (1.17), respectively. Quantitative meta-analysis, using a random-effects model, determined the overall pooled flea prevalence in the Middle East as 40% (95% CI: 25-55, I2 = 100%, p < 0.00001), ranging between 13% (95% CI: 0-30, I2 = 95%, p < 0.00001) in Iran and 59% (95% CI: 42-77, I2 = 75%, p < 0.00001) in Israel. The overall pooled louse prevalence was found to be 30% (95% CI: 13-47, I2 = 100%, p < 0.00001), ranging between 25% in Iran (95% CI: 1-50, I2 = 99%) and 38% in Egypt (95% CI: 7-68, I2 = 100%). In the case of mites, the pooled prevalence in this region was 33% (95% CI: 11-55, I2 = 100%, p < 0.00001), where the country-specific prevalence estimates were 30% in Iran (95% CI: 4-56, I2 = 99%) and 32% in Egypt (95% CI: 0-76, I2 = 100%). For ticks, the overall prevalence was found to be 25% (95% CI: 2-47, I2 = 100%, p < 0.00001), ranging from 16% in Iran (95% CI: 7-25, I2 = 74%) to 42% in Egypt (95% CI: 1-85, I2 = 100%). The control of rodent ectoparasites should be considered to reduce their adverse effects. Using the One Health strategy, rodent control, and precisely control of the most common rodent species, i.e., Mus musculus, Rattus norvegicus, and Rattus rattus, should be considered to control the rodent-borne ectoparasites in this region.


The Association Between Cancer and Spousal Rate of Memory Decline: A Negative Control Study to Evaluate (Unmeasured) Social Confounding of the Cancer-memory Relationship.
Cancer diagnoses are associated with better long-term memory in older adults, possibly reflecting a range of social confounders that increase cancer risk but improve memory. We used spouse's memory as a negative control outcome to evaluate this possible confounding, since spouses share social characteristics and environments, and individuals' cancers are unlikely to cause better memory among their spouses. We estimated the association of an individual's incident cancer diagnosis (exposure) with their own (primary outcome) and their spouse's (negative control outcome) memory decline in 3601 couples from 1998 to 2014 in the Health and Retirement Study, using linear mixed-effects models. Incident cancer predicted better long-term memory for the diagnosed individual. We observed no association between an individual's cancer diagnosis and rate of spousal memory decline. This negative control study suggests that the inverse association between incident cancer and rate of memory decline is unlikely to be attributable to social/behavioral factors shared between spouses.


NMR constrained solution structures for laminin peptide 11. Analogs define structural requirements for inhibition of tumor cell invasion of basement membrane matrix.
Peptide 11, CDPGYIGSR-NH2, is a segment of laminin which blocks tumor cell invasion. A high affinity laminin receptor in tumor cells is thought to be blocked by the carboxyl-terminal YIGSR, and conformational energy calculations suggest that the glycine in YIGSR allows an important conformational bend. We replaced the YIGSR glycine residue in peptide 11 with either D-alanine or L-alanine to allow or disfavor the proposed glycine bend. We found the Gly7-->D-Ala7 analog to be equal to peptide 11 in inhibiting tumor cell invasion of basement membrane matrix. The Gly7-->L-Ala7 analog was much less capable of invasion inhibition. Two-dimensional 1H-1H NMR was used to study the solution conformations of the peptide 11 analogs. NOESY experiments revealed close NH-NH contacts in peptide 11 and the D-Ala7 analog, but not in the L-Ala7 analog. Molecular dynamics generated low energy structures with excellent NOE agreement for peptide 11 and its analogs. Both peptide 11 and the D-Ala7 analog, but not the less active L-Ala7 analog, were predicted to have similar bends around Gly7 or D-Ala7. These results suggest that a bend in the YIGSR region of peptide 11 may be important for the binding of laminin to its metastasis-associated receptor.


Short-term patient-reported outcomes following concomitant hip arthroscopy and the endoscopic modified shelf procedure for the treatment of acetabular dysplasia and intra-articular pathology.
The shelf procedure is a treatment of acetabular dysplasia, with the aim of increasing weight-bearing acetabular coverage. Although several shelf techniques have been described, the endoscopic procedure with concomitant hip arthroscopy is a new, less invasive alternative. Outcomes following this procedure are scarce. The purpose of this study was to report short-term patient-reported outcomes (PROs) following concomitant hip arthroscopy and endoscopic modified shelf procedure in the setting of acetabular dysplasia and labral tears. Patients that met extraordinarily selective surgical indications and underwent the abovementioned surgery between February 2016 and October 2019 and had minimum 1-year follow-up were included. There were five females with a mean age of 40.18 ± 5.05 years and follow-up of 21.55 ± 8.68 months. The lateral center-edge angle increased from 15.80° to 23.20° (P = 0.003), and vertical center-edge angle increased from 16.60° to 23.60° (P < 0.001). The Tönnis angle decreased from 15.40° to 3.74° (P < 0.001). The alpha angle decreased from 58.46° to 40.70° (P < 0.001). PROs demonstrated significant improvement at latest follow-up (modified Harris Hip Score, P = 0.042; Non-Arthritic Hip Score, P < 0.001; Hip Outcome Score-Sports Specific Subscale, P = 0.035; Visual Analog Scale, P < 0.001; International Hip Outcome Tool-12, P = 0.043), and satisfaction was 8.60 ± 0.89. No secondary surgeries were reported. Concomitant hip arthroscopy and endoscopic modified shelf procedure appears to be a safe and effective procedure for patients with acetabular dysplasia and labral tears yielding favorable outcomes and satisfaction at short-term follow-up.


Preferences of patients and clinicians for treatment of Graves' disease: a discrete choice experiment.
Treatment options for Graves' disease (GD) consist of antithyroid drugs (ATD), radioactive iodine (RAI) and total thyroidectomy (TT). Guidelines recommend to discuss these options with patients, taking into account patients' preferences. This study aims to evaluate and compare patients' and clinicians' preferences and the trade-offs made in choosing treatment. A discrete choice experiment (DCE) was performed with GD patients with a first diagnosis or recurrence in the previous year, and with clinicians. Participants were offered hypothetical treatment options which differed in type of treatment, rates of remission, severe side effects, permanent voice changes and hypocalcemia. Preference heterogeneity was assessed by latent-class analysis. In this study, 286 (82%) patients and 61 (18%) clinicians participated in the DCE. All treatment characteristics had a significant effect on treatment choice (P < 0.05). Remission rate was the most important determinant and explained 37 and 35% of choices in patients and clinicians, respectively. Both patients and clinicians preferred ATD over surgery and RAI. A strong negative preference toward RAI treatment was observed in a subclass of patients, whereas clinicians preferred RAI over surgery. For both patients and clinicians, remission rate was the most important determinant of treatment choice and ATD was the most preferred treatment option. Patients had a negative preference toward RAI compared to alternatives, whereas clinicians preferred RAI over surgery. Clinicians should be aware that their personal attitude toward RAI differs from that of their patients. This study on patients' and clinicians' preferences can support shared decision making and thereby improve clinical treatment.


Study of miRNA interactome in active rheumatoid arthritis patients reveals key pathogenic roles of dysbiosis in the infection-immune network.
To characterize serum microRNA (miR) and the miR interactome of active RA patients in RA aetiology and pathogenesis. The differentially expressed miRs (DEmiRs) in serum of naïve active RA patients (NARAPs, n = 9, into three pools) vs healthy controls (HCs, n = 15, into five pools) were identified with Agilent human miR microarray analysis. Candidate driver genes in epigenetic and pathogenic signalling pathway modules for RA were analysed using miRTarBase and a molecular complex detection algorithm. The interactome of these DEmiRs in RA pathogenesis were further characterized with gene ontology and Kyoto Encyclopaedia of Genes and Genomes. Three upregulated DEmiRs (hsa-miR-187-5p, -4532, -4516) and eight downregulated DEmiRs (hsa-miR-125a-3p, -575, -191-3p, -6865-3p, -197-3p, -6886-3p, -1237-3p, -4436b-5p) were identified in NARAPs. Interactomic analysis from heterogeneous experimentally validated sources yielded 1719 miR-target interactions containing 5.67% strong and 94.33% less strong experimental evidence. Gene ontology and Kyoto Encyclopaedia of Genes and Genomes analyses allocated the upregulated DEmiRs in the infection modules and the downregulated DEmiRs in the immune signalling pathways. Specifically, these DEmiRs revealed the significant contributions of the intestinal microbiome dysbiosis in the infection-inflammation-immune network for activation of T cells, immune pathways of IL-17, Toll-like receptor, TNF, Janus kinase-signal transducer and activator of transcription, osteoclast cell differentiation pathway and IgA production to the active RA pathogenesis. Our experiment-based interactomic study of DEmiRs in serum of NARAPs revealed novel clinically relevant miRs interactomes in the infection-inflammation-immune network of RA. These results provide valuable resources for understanding the integrated function of the miR network in RA pathogenesis and the application of circulating miRs as biomarkers for early aetiologic RA diagnosis.


Osteoblast-Osteoclast Communication and Bone Homeostasis.
Bone remodeling is tightly regulated by a cross-talk between bone-forming osteoblasts and bone-resorbing osteoclasts. Osteoblasts and osteoclasts communicate with each other to regulate cellular behavior, survival and differentiation through direct cell-to-cell contact or through secretory proteins. A direct interaction between osteoblasts and osteoclasts allows bidirectional transduction of activation signals through EFNB2-EPHB4, FASL-FAS or SEMA3A-NRP1, regulating differentiation and survival of osteoblasts or osteoclasts. Alternatively, osteoblasts produce a range of different secretory molecules, including M-CSF, RANKL/OPG, WNT5A, and WNT16, that promote or suppress osteoclast differentiation and development. Osteoclasts also influence osteoblast formation and differentiation through secretion of soluble factors, including S1P, SEMA4D, CTHRC1 and C3. Here we review the current knowledge regarding membrane bound- and soluble factors governing cross-talk between osteoblasts and osteoclasts.


Personal experience on treatment of colorectal liver metastases: a multidisciplinary approach.
Diagnosed colorectal cancer patients presents liver metastases at presentation in 25% of cases, an another 25% of patients will develop hepatic recurrence during follow-up period. Natural history of disease in untreated patients is sad without long term survivors. The only chance of curative treatment is surgical resection. Aim of this study was to evaluate recurrence and survival in the multimodal approach of colorectal liver metastases patients. From 1990 to 2003, 212 colorectal cancer liver metastases patients were observed at the "Regina Elena" cancer institute of Rome, the 48.8% of cases were synchronous. Of 185 patients with minimum follow-up of 12 months, 80 were surgically resected, while 105 were not. Major resection was possible in 45.3% of cases and 66.1% of them were radicals. Tweny were treated by surgery alone, 65% were resected after i.v. chemotherapy, in the last 10% resection followed HAI chemotherapy. Of non resected patients, in the 36.2% i.v. chemotherapy was administered, 26.1% were submitted to HAI chemotherapy, in 18.9% others treatments were carried out and 18.8 were untreated. Operative mortality was contained to 2.5%. The hepatic recurrence was observed in 50.5% of cases while extrahepatic was in the 13.6%. In the resected cases the 5-yr. Overall survival was 38.8%, the disease-free survival decreased to 14.7% with a median time to progression of 15 months. The liver recurrence-free survival was 31.3%. Really bad prognopsis was observed in the non resected patients with none survived at 5 years and 3-yr overall survival of 18.7%. In selected patients the multimodality approach to treatment of hepatic metastases showed satisfactory results in terms of local and distal control of disease. Long term survivors were observed in the our experience.


Three-dimensional structure of favin: saccharide binding-cyclic permutation in leguminous lectins.
The three-dimensional structure of favin, the glucose- and mannose-binding lectin of Vicia faba (vetch, broad bean), has been determined at a resolution of 2.8 angstroms by molecular replacement. The crystals contain specifically bound glucose and provide the first high-resolution view of specific saccharide binding in a leguminous lectin. The structure is similar to those of concanavalin A (Con A) and green pea lectin; differences from Con A show that minimal changes are needed to accommodate the cyclic permutation in amino acid sequence between the two molecules. The molecule is an ellipsoidal dimer dominated by extensive beta structures. Each protomer contains binding sites for two divalent metal ions (Mn2+ and Ca2+) and a specific saccharide. Glucose is bound by favin in a cleft in the molecular surface and has noncovalent contacts primarily with two peptide loops, one of which contains several metal ion ligands. The specific carbohydrate-binding site is similar to that of Con A in location and general peptide folding, despite several differences in specific amino acid residues.


Collaborative-controlled LASSO for constructing propensity score-based estimators in high-dimensional data.
Propensity score-based estimators are increasingly used for causal inference in observational studies. However, model selection for propensity score estimation in high-dimensional data has received little attention. In these settings, propensity score models have traditionally been selected based on the goodness-of-fit for the treatment mechanism itself, without consideration of the causal parameter of interest. Collaborative minimum loss-based estimation is a novel methodology for causal inference that takes into account information on the causal parameter of interest when selecting a propensity score model. This "collaborative learning" considers variable associations with both treatment and outcome when selecting a propensity score model in order to minimize a bias-variance tradeoff in the estimated treatment effect. In this study, we introduce a novel approach for collaborative model selection when using the LASSO estimator for propensity score estimation in high-dimensional covariate settings. To demonstrate the importance of selecting the propensity score model collaboratively, we designed quasi-experiments based on a real electronic healthcare database, where only the potential outcomes were manually generated, and the treatment and baseline covariates remained unchanged. Results showed that the collaborative minimum loss-based estimation algorithm outperformed other competing estimators for both point estimation and confidence interval coverage. In addition, the propensity score model selected by collaborative minimum loss-based estimation could be applied to other propensity score-based estimators, which also resulted in substantive improvement for both point estimation and confidence interval coverage. We illustrate the discussed concepts through an empirical example comparing the effects of non-selective nonsteroidal anti-inflammatory drugs with selective COX-2 inhibitors on gastrointestinal complications in a population of Medicare beneficiaries.


How thyroid disease presents in the elderly.
Some of the symptoms and signs of hypothyroidism and hyperthyroidism in elderly patients may be mistakenly attributed to "old age." Weight loss, muscle weakness, tremor, angina, congestive heart failure--all signs of hyperthyroidism--are also concomitants of aging. Fatigue, sluggishness, withdrawal behavior, senile atrophic skin changes--all signs of hypothroidism--are also a part of the normal aging process. Although screening elderly people for thyroid disease is economically unsound, the physician should maintain a high index of suspicion of its presence. Laboratory tests must be interpreted with extra care. Values of 131I uptake, serum T4 and T3, thyroid-stimulating hormone, and thyrotropin-releasing hormone are all helpful in diagnosis. Thyroid disease is easily treated in elderly patients, and results often are dramatic. Propranolol is effective in thyrotoxic patients when symptoms require prompt relief. The definitive treatment, however, is 131I; antithyroid drugs are difficult to manage. Hypothyroidism is easily treated with T4.


Effects of Acute Alcohol Exposure on Layer 5 Pyramidal Neurons of Juvenile Mice.
Early-onset drinking during childhood or preadolescence is a serious social problem. Yet, most of the basic neurobiological research on the acute effects of ethanol has been carried out on adult or early postnatal animals. We studied the effect of alcohol exposure on the basic electrophysiological properties and cell viability of layer 5 pyramidal neurons from the somatosensory cortex of juvenile (P21-P23) C57BL/6N mice. After bath application of 50 mM ethanol to acute slices of the somatosensory cortex, no adverse effects were detected on cells survival, whereas the input resistance and firing rate of layer 5 neurons were significantly reduced. While the effect on the input resistance was reversible, the depressing effect on cell firing remained stable after 6 min of alcohol exposure. Ethanol application did not result in any significant change of mIPSC frequency, amplitude, and rise time. A slight increase of mIPSC decay time was observed after 6 min of ethanol exposure. The molecular mechanisms leading to these alterations and their significance for the physiology of the cerebral cortex are briefly discussed.


[Cost-effectiveness of a TLC-NOSF polyurethane foam dressing].
Chronic wounds represent a drain on the Spanish health system, nowdays is necessary an optimization of the resources used and that is for this that is necessary justify the use of the products over others through cost-effective studies for to show the economic benefit to professionals and the life quality of patient. This article compares the use of a new technology for format polyurethane foam, TLC-NOSF, with the most commonly used products for treating wounds. This comparison is made using a cost-effectiveness model (Markov Model). The results demonstrate that treatment with polyurethane foam dressing with TLC-NOSF are cost-effective versus treatments with polyurethane foams most commonly used in Spain.


Spectrum of Phenotypic, Genetic, and Functional Characteristics in Patients With Epilepsy With KCNC2 Pathogenic Variants.
KCNC2 encodes Kv3.2, a member of the Shaw-related (Kv3) voltage-gated potassium channel subfamily, which is important for sustained high-frequency firing and optimized energy efficiency of action potentials in the brain. The objective of this study was to analyze the clinical phenotype, genetic background, and biophysical function of disease-associated Kv3.2 variants. Individuals with KCNC2 variants detected by exome sequencing were selected for clinical, further genetic, and functional analysis. Cases were referred through clinical and research collaborations. Selected de novo variants were examined electrophysiologically in Xenopus laevis oocytes. We identified novel KCNC2 variants in 18 patients with various forms of epilepsy, including genetic generalized epilepsy (GGE), developmental and epileptic encephalopathy (DEE) including early-onset absence epilepsy, focal epilepsy, and myoclonic-atonic epilepsy. Of the 18 variants, 10 were de novo and 8 were classified as modifying variants. Eight drug-responsive patients became seizure-free using valproic acid as monotherapy or in combination, including severe DEE cases. Functional analysis of 4 variants demonstrated gain of function in 3 severely affected DEE cases and loss of function in 1 case with a milder phenotype (GGE) as the underlying pathomechanisms. These findings implicate KCNC2 as a novel causative gene for epilepsy and emphasize the critical role of KV3.2 in the regulation of brain excitability.


IVF and ICSI in male infertility: update on outcomes, risks, and costs.
Assisted reproductive technology with intracytoplasmic sperm injection (ICSI) is becoming an international panacea for couples struggling with infertility. The increasing popularity of these techniques and the data generated has given us a better understanding of the efficacy, consequences and costs of these procedures. There still remain many unanswered questions and controversies surrounding the use of IVF and ICSI. Increased experience, better refinement of these techniques and clearer indications for IVF and ICSI will inevitably minimize the risks associated with this procedure.


Genomic variation within the maize stiff-stalk heterotic germplasm pool.
The stiff-stalk heterotic group in Maize (Zea mays L.) is an important source of inbreds used in U.S. commercial hybrid production. Founder inbreds B14, B37, B73, and, to a lesser extent, B84, are found in the pedigrees of a majority of commercial seed parent inbred lines. We created high-quality genome assemblies of B84 and four expired Plant Variety Protection (ex-PVP) lines LH145 representing B14, NKH8431 of mixed descent, PHB47 representing B37, and PHJ40, which is a Pioneer Hi-Bred International (PHI) early stiff-stalk type. Sequence was generated using long-read sequencing achieving highly contiguous assemblies of 2.13-2.18 Gbp with N50 scaffold lengths >200 Mbp. Inbred-specific gene annotations were generated using a core five-tissue gene expression atlas, whereas transposable element (TE) annotation was conducted using de novo and homology-directed methodologies. Compared with the reference inbred B73, synteny analyses revealed extensive collinearity across the five stiff-stalk genomes, although unique components of the maize pangenome were detected. Comparison of this set of stiff-stalk inbreds with the original Iowa Stiff Stalk Synthetic breeding population revealed that these inbreds represent only a proportion of variation in the original stiff-stalk pool and there are highly conserved haplotypes in released public and ex-Plant Variety Protection inbreds. Despite the reduction in variation from the original stiff-stalk population, substantial genetic and genomic variation was identified supporting the potential for continued breeding success in this pool. The assemblies described here represent stiff-stalk inbreds that have historical and commercial relevance and provide further insight into the emerging maize pangenome.


Chemical bonding in pentaerythritol at very low temperature or at high pressure: an experimental and theoretical study.
Chemical bonding in the pentaerythritol crystal based on the experimental electron density at 15 (1) K, and theoretical calculations at the experimental molecular geometries obtained at room and low (15 K) temperatures have been analyzed and compared in terms of the topological analysis. Topological electron-density features corresponding to the high-pressure (1.15 GPa) geometry are also reported. In addition to the bond critical points (CPs) within the molecular layers, CPs between the atoms of different molecular layers have been located and the bonding character of these relatively weak interactions discussed. Atomic charges and energies have been integrated over the atomic basins delimited by the zero-flux surfaces, and the intermolecular interaction energies have been calculated. The interaction between molecular layers in the crystal becomes stronger both at very low temperature and high pressure, as demonstrated by the more negative intermolecular interaction energies, higher electron density and energy density values at the CPs, and sharper electronic-energy density profiles.


Characterization of a small vesicular cholesterol carrier in human gallbladder bile.
Cholesterol phospholipid vesicles play an important role in the nucleation of cholesterol in bile. Recent studies have identified an additional vesicle population in human bile. In this study, the role of these small vesicles as cholesterol carriers was examined. Gallbladder bile was obtained from 60 patients at cholecystectomy. Large vesicles, small vesicles, lamellae, and mixed micelles were separated using gel filtration chromatography. Small vesicles were present in bile from the majority of patients both with and without cholesterol gallstones, whereas the void volume vesicle fraction was found almost exclusively in bile from patients with cholesterol gallstones. Both large vesicular and small vesicular cholesterol increased as total bile cholesterol concentration increased; however, the cholesterol-phospholipid ratio in the large vesicle fraction from patients with cholesterol stones was significantly greater than the ratio in small vesicles (1.6 +/- 0.3 vs. 1.0 < or = 0.1, p < 0.05). Whole bile cholesterol crystal appearance time was correlated significantly with the percentage of cholesterol transported by large vesicles (r = 0.63, p < 0.001) but not with the percentage of cholesterol present in small vesicles. Finally, large vesicles isolated by gel filtration chromatography formed cholesterol crystals faster than small vesicles (5.3 +/- 2 vs. 17.4 +/- 4 days, p < 0.01). These data suggest that a heterogenous population of vesicles is present in human gallbladder bile. As bile becomes saturated with cholesterol, it increasingly is solubilized by both small and large vesicles. The small vesicles have relatively less cholesterol and are more stable than the larger variety, from which cholesterol is most likely to precipitate.


The stability of family decisions to consent or refuse organ donation: would you do it again?
Past organ donation research has studied attitudes toward donation, predictors of signing donor cards, and distinguishing characteristics of donors vs. nondonors. The current study is the first to examine predictors of family members' satisfaction with the decision to consent or refuse donation of a dying loved one's organs or tissue. This study surveyed 225 family members who had been approached to donate the organs or tissue of a dying loved one. Participants were surveyed about demographic characteristics, medical/hospital factors, previous knowledge of transplantation, the request process, religion, and characteristics of the deceased and of the recipient. Discriminant analyses were conducted to characterise four specific groups: a) donors who would donate again; b) donors who would not donate again; c) nondonors who would now donate; and d) nondonors who still would not donate. Three significant discriminant functions emerged discriminating donors from nondonors, those who were satisfied with their decision from those who were not, and people who would now donate from those who would not. Donation was predicated by formal education, being married, volunteerism, signing donor cards, and having personal conversations about donation. Subsequent satisfaction was predicted by comfort and confidence during the decision-making process, familiarity with medical center, and understanding of brain death. A willingness to now donate was predicted by personal discussions about donation. People should be encouraged not only to sign donor cards, but to have discussions with family about wishes. Individuals should be encouraged to seek the help of family and friends during the decision, and be aware of the need of social support from family and friends during and after the decision.


Patellofemoral Pain Syndrome Modifies the Movement of the Rearfoot, but It Does Not Alter Plantar Pressure Distribution.
Objective To compare the plantar pressure distribution and the kinematics of the rearfoot on the stance phase of subjects with or without patellofemoral pain syndrome (PFPS). Methods A total of 26 subjects with PFPS and 31 clinically healthy subjects, who were paired regarding age, height and mass, participated in the study. The plantar pressure distribution (peak pressure) was assessed in six plantar regions, as well as the kinematics of the rearfoot (maximum eversion angle, percentage of the stance phase when the maximum angle was reached, and percentage of the stance phase in which the rearfoot was in eversion). The data were analyzed by descriptive and inferential statistics, with a significance level of p ≤ 0.05. Results The pressure on the six plantar regions analyzed and the magnitude of the maximum eversion angle of the rearfoot when walking on flat surfaces did not present differences among the subjects with PFPS. However, the PFPS subjects showed, when walking, an earlier maximum eversion angle of the rearfoot than the subjects on the control group, and stayed less time with the rearfoot in eversion. Conclusion The PFPS seems to be related to modifications on the temporal pattern on the kinematics of the rearfoot.


Capsule endoscopy and enteroscopy: modern modalities to investigate the small bowel in paediatrics.
Historically the small bowel has been considered a technically difficult area to examine because of its length (3-5 metres), location and tortuosity. Capsule endoscopy and enteroscopy have revolutionised the investigation pathway of the small bowel in adults. They are now developing increasingly important roles as modalities of investigation in paediatrics. This review appraises the current literature to define the clinical indications and practical aspects of capsule endoscopy and enteroscopy that are of interest to the clinician.


BRAF status modulates Interelukin-8 expression through a CHOP-dependent mechanism in colorectal cancer.
Inflammation might substantially contribute to the limited therapeutic success of current systemic therapies in colorectal cancer (CRC). Amongst cytokines involved in CRC biology, the proinflammatory chemokine IL-8 has recently emerged as a potential prognostic/predictive biomarker. Here, we show that BRAF mutations and PTEN-loss are associated with high IL-8 levels in CRC models in vitro and that BRAF/MEK/ERK, but not PI3K/mTOR, targeting controls its production in different genetic contexts. In particular, we identified a BRAF/ERK2/CHOP axis affecting IL-8 transcription, through regulation of CHOP subcellular localization, and response to targeted inhibitors. Moreover, RNA Pol II and an open chromatin status in the CHOP-binding region of the IL-8 gene promoter cooperate towards increased IL-8 expression, after a selective BRAF inhibition. Overall, our data show that IL-8 production is finely and differentially regulated depending on the tumor genetic context and might be targeted for therapeutic purposes in molecularly defined subgroups of CRC patients.


Apolipoprotein AI levels are increased in part as a consequence of reduced catabolism in nephrotic rats.
Apolipoprotein AI (apo AI) synthesis, measured as the turnover of 125I-labeled apo AI-labeled high-density lipoprotein (HDL), was increased significantly in rats with Heymann nephritis (HN) vs. control Sprague-Dawley (SD) rats. However, fractional apo AI catabolic rate was also significantly less in HN vs. SD. We used 125I-apo AI tyramine cellobiose HDL, a marker retained at the catabolic site, to establish where apo AI catabolism decreased in six HN rats, seven rats with adriamycin (Adria)-induced nephrosis, and six control SD. Total renal apo AI catabolism, plus urinary losses, were the same in all three groups, despite significant urinary apo AI in HN and Adria rats. Apo AI catabolism was reduced in skin in both nephrotic groups, accounting for approximately 44% of reduced in apo AI catabolism. Thus a significant fraction of apo AI is catabolized in skin of normal male rats. Reduced apo AI catabolism in skin contributes to increased plasma levels in nephrotic rats.


Primary perineuritis, a rare but treatable neuropathy: Review of perineurial anatomy, clinicopathological features, and differential diagnosis.
The perineurium surrounds each fascicle in peripheral nerves, forming part of the blood-nerve barrier. We describe its normal anatomy and function. "Perineuritis" refers to both a nonspecific histopathological finding and more specific clinicopathological entity, primary perineuritis (PP). Patients with PP are often assumed to have nonsystemic vasculitic neuropathy until nerve biopsy is performed. We systematically reviewed the literature on PP and developed a differential diagnosis for histopathologically defined perineuritis. We searched PubMed, Embase, Scopus, and Web of Science for "perineuritis." We identified 20 cases (11 M/9F) of PP: progressive, unexplained neuropathy with biopsy showing perineuritis without vasculitis or other known predisposing condition. Patients ranged in age from 18 to 75 (mean 53.7) y and had symptoms 2-24 (median 4.5) mo before diagnosis. Neuropathy was usually sensory-motor (15/20), painful (18/19), multifocal (16/20), and distal-predominant (16/17) with legs more affected than arms. Truncal numbness occurred in 6/17; 10/18 had elevated cerebrospinal fluid (CSF) protein. Electromyography (EMG) and nerve conduction studies (NCS) demonstrated primarily axonal changes. Nerve biopsies showed T-cell-predominant inflammation, widening, and fibrosis of perineurium; infiltrates in epineurium in 10/20 and endoneurium in 7/20; and non-uniform axonal degeneration. Six had epithelioid cells. 19/20 received corticosteroids, 8 with additional immunomodulators; 18/19 improved. Two patients did not respond to intravenous immunoglobulin (IVIg). At final follow-up, 13/16 patients had mild and 2/16 moderate disability; 1/16 died. Secondary causes of perineuritis include leprosy, vasculitis, neurosarcoidosis, neuroborreliosis, neurolymphomatosis, toxic oil syndrome, eosinophilia-myalgia syndrome, and rarer conditions. PP appears to be an immune-mediated, corticosteroid-responsive disorder. It mimics nonsystemic vasculitic neuropathy. Cases with epithelioid cells might represent peripheral nervous system (PNS)-restricted forms of sarcoidosis.


Modeling the Etiology of Adolescent Substance Use: A Test of the Social Development Model.
The social development model is a general theory of human behavior that seeks to explain antisocial behaviors through specification of predictive developmental relationships. It incorporates the effects of empirical predictors ("risk factors" and "protective factors") for antisocial behavior and attempts to synthesize the most strongly supported propositions of control theory, social learning theory, and differential association theory. This article examines the power of social development model constructs measured at ages 9 to 10 and 13 to 14 to predict drug use at ages 17 to 18. The sample of 590 is from the longitudinal panel of the Seattle Social Development Project, which in 1985 sampled fifth grade students from high crime neighborhoods in Seattle, Washington. Structural equation modeling techniques were used to examine the fit of the model to the data. Although all but one path coefficient were significant and in the expected direction, the model did not fit the data as well as expected (CFI=.87). We next specified second-order factors for each path to capture the substantial common variance in the constructs' opportunities, involvement, and rewards. This model fit the data well (CFI=.90). We conclude that the social development model provides an acceptable fit to predict drug use at ages 17 to 18. Implications for the temporal nature of key constructs and for prevention are discussed.


Progressive Multifocal Leukoencephalopathy Diagnosed by Brain Biopsy, not by the DNA Test for JC Virus.
Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by a lytic infection of oligodendrocytes due to the presence of JC polyomavirus (JCV). The disease occurs mostly in immunocompromised patients and is associated with a high mortality rate. The diagnosis of PML is based on a polymerase chain reaction (PCR) assay for JC viral DNA in cerebrospinal fluid (CSF). However, case reports of the diagnosis of PML established with brain biopsy despite negative JCV CSF PCR analysis when clinical and neuroimaging features are suggestive of PML have been published. A 44-year-old male with a 6-year history of acquired immunodeficiency syndrome developed mental confusion and memory impairment despite 3 months of highly active antiretroviral therapy. Magnetic resonance imaging revealed multiple subcortical white matter lesions in bilateral hemispheres and subcortical nuclei including the thalamus and basal ganglia. JCV DNA was not detected in CSF study, but a brain biopsy showed a high JCV DNA titer. The diagnosis of PML was established with brain biopsy. An early brain biopsy may be important in the diagnosis of PML in patients with clinical manifestations and neuroimaging findings if JCV DNA is undetectable in the CSF PCR.


Sequence analysis of p44 homologs expressed by Anaplasma phagocytophilum in infected ticks feeding on naive hosts and in mice infected by tick attachment.
The 44-kDa immunodominant outer membrane proteins (P44 proteins) of Anaplasma phagocytophilum are encoded by the p44 polymorphic multigene family. The present study examined p44 expression and analyzed the cDNA sequences of various p44 transcripts from the spleens and blood of mice infected by the bites of ticks infected with the A. phagocytophilum NTN-1 strain or of naturally infected nymphal ticks and in the salivary glands and midgut tissues of these ticks. A total of 300 p44 cDNAs were subjected to sequence analysis. Of these, 40 distinct p44 species were found, and all of these had orthologs in the A. phagocytophilum HZ strain genome that shared 95 to 100% base sequence identity. The number of unique p44 species expressed in mouse blood was greater than that for mouse spleens. Higher numbers of different p44 transcripts were also expressed in the salivary glands of ticks than in the midgut tissues. Variations in the sequences of the same p44 cDNA species within a single A. phagocytophilum strain and among different strains were concentrated in the conserved regions flanking the central hypervariable region of p44 genes. No mosaic sequences derived from two or more p44 species were found within the p44 hypervariable region. The conservation of the hypervariable region of each p44 cDNA species of A. phagocytophilum in naturally infected ticks and in different geographic isolates suggests that each A. phagocytophilum genome carries a set of p44 paralogs to be expressed. Thus, a large but restricted repertoire of p44 hypervariable sequences exists in A. phagocytophilum strains in the Northeastern United States.


The potential impact of the National Osteoporosis Foundation guidance on treatment eligibility in the USA: an update in NHANES 2005-2008.
This analysis of National Health and Nutrition Examination Survey (NHANES) 2005-2008 data describes the prevalence of risk factors for osteoporosis and the proportions of men and postmenopausal women age 50 years and older who are candidates for treatment to lower fracture risk, according to the new Fracture Risk Assessment Tool (FRAX)-based National Osteoporosis Foundation Clinician's Guide. It is important to update estimates of the proportions of the older US population considered eligible for pharmacologic treatment for osteoporosis for purposes of understanding the health care burden of this disease. This is a cross-sectional study of the NHANES 2005-2008 data in 3,608 men and women aged 50 years and older. Variables in the analysis included race/ethnicity, age, lumbar spine and femoral neck bone mineral density, risk factor profiles, and FRAX 10-year fracture probabilities. The prevalence of osteoporosis of the femoral neck ranged from 6.0% in non-Hispanic black to 12.6% in Mexican American women. Spinal osteoporosis was more prevalent among Mexican American women (24.4%) than among either non-Hispanic blacks (5.3%) or non-Hispanic whites (10.9%). Treatment eligibility was similar in Mexican American and non-Hispanic white women (32.0% and 32.8%) and higher than it was in non-Hispanic black women (11.0%). Treatment eligibility among men was 21.1% in non-Hispanic whites, 12.6% in Mexican Americans, and 3.0% in non-Hispanic blacks. Nineteen percent of older men and 30% of older women in the USA are at sufficient risk for fracture to warrant consideration for pharmacotherapy.


How should we sequence and combine novel therapies in CLL?
With the recent approval of several effective and well-tolerated novel agents (NAs), including ibrutinib, idelalisib, venetoclax, and obinutuzumab, patients with chronic lymphocytic leukemia (CLL) have more therapeutic options than ever before. The availability of these agents is both an important advance for patients but also a challenge for practicing hematologist/oncologists to learn how best to sequence NAs, both with respect to chemoimmunotherapy (CIT) and to other NAs. The sequencing of NAs in clinical practice should be guided both by an individual patient's prognostic markers, such as FISH and immunoglobulin heavy chain variable region (IGHV)-mutation status, as well as the patient's medical comorbidities and goals of care. For older, frailer patients with lower-risk CLL prognostic markers, NA monotherapy may remain a mainstay of CLL treatment for years to come. For younger, fitter patients and those with higher-risk CLL, such as del(17p) or unmutated IGHV, combination approaches may prove to be more valuable than NA monotherapy. Trials are currently evaluating the efficacy of several such combination approaches, including NA plus anti-CD20 monoclonal antibody, NA plus NA (with or without anti-CD20 monoclonal antibody), and NA plus CIT. Given the tremendous efficacy of the already approved NAs, as well as the promising data for next generation NAs, the development of well-tolerated, highly effective combination strategies with curative potential for patients with CLL has become a realistic goal.


Prenatal androgen influences on the brain: A review, critique, and illustration of research on congenital adrenal hyperplasia.
Sex hormones, especially androgens, contribute to sex and gender differences in the brain and behavior. Organizational effects are particularly important because they are thought to be permanent, reflecting hormone exposure during sensitive periods of development. In human beings, they are often studied with natural experiments in which sex hormones are dissociated from other biopsychosocial aspects of development, such as genes and experiences. Indeed, the greatest evidence for organizational effects on sex differences in human behavior comes from studies of females with congenital adrenal hyperplasia (CAH), who have heightened prenatal androgen exposure, female-typical rearing, and masculinized toy play, activity and career interests, spatial skills, and some personal characteristics. Interestingly, however, neuroimaging studies of females with CAH have revealed few neural mechanisms underlying these hormone-behavior links, with the exception of emotion processing; studies have instead shown reduced gray matter volumes and reduced white matter integrity most consistent with other disease-related processes. The goals of this narrative review are to: (a) describe methods for studying prenatal androgen influences, while offering a brief overview of behavioral outcomes; (b) provide a critical methodological review of neuroimaging research on females with CAH; (c) present an illustrative analysis that overcomes methodological limitations of previous work, focusing on person-specific neural reward networks (and their associations with sensation seeking) in women with CAH and their unaffected sisters in order to inform future research questions and approaches that are most likely to reveal organizational hormone effects on brain structure and function.


Effect of a Mineral-Microbial Deodorizing Preparation on the Functions of Internal Organs and the Immune System in Commercial Poultry.
Animal production is identified as one of the main sources of high concentrations of odours, which are related to air pollution, health problems of living organisms and indirect negative impact on production results. One common method for reducing emissions of ammonia is using preparations containing probiotics and hygroscopic or disinfecting compounds. This study was undertaken in order to determine the impact of innovative mineral-microbial deodorizing preparation, which reduces odorous gases, applying to the litter once a week in poultry houses on the physiological status of breeder chickens, broiler chickens and turkeys. Samples were collected after slaughter and analyzed using ELISA tests, flow cytometry and biochemical methods. Biochemical markers of the liver and kidney profile (ALT, AST, LDH, ALP, CK, TP, CALC, PHOS) and the titers of specific antibodies against AEV, aMPV, AAvV-1, IBDV, HEV, BA were analyzed in serum samples. The percentage contribution of T and B lymphocyte subpopulations was determined in the samples of tracheal mucosa, blood, and spleen. No significant differences were found between the control and experimental group with regard to all the analyzed parameters, with some exceptions for biochemistry. The results of our study indicated that mineral-microbial deodorizing preparation did not affect the physiological status of birds.