Study of Dickkopf-1 (DKK-1) Gene Expression in Hepatocellular Carcinoma Patients.
Hepatocellular Carcinoma (HCC) is the sixth most common cancer in the world. Dickkopf -1 (DKK-1) protein is a new biomarker used in conjunction with Alpha Fetoprotein (AFP) to differentiate HCC from "non-malignant" liver disease. DKK-1 is an inhibitor of Wnt/β-catenin signaling pathway which is involved in embryogenesis and has been implicated in tumorigenesis in many tissues. To investigate the level of DKK-1 gene expression in the peripheral blood of patients with HCC who had a history of Hepatitis C Virus (HCV) and schistosomal infections. This "cross-sectional" study was carried out in the Tropical Medicine Department of Tanta University Hospital on 50 patients with HCC and 10 healthy volunteers served as control. All patients were tested for HCV antibodies and "anti-schistosomal" antibodies. All groups were tested for DKK-1 gene expression which was measured with quantitative real-time PCR. DKK-1 gene was over-expressed in HCC patients than in the control group with mean 3.269±4.762 versus 1.00 in controls (p< 0.005). "Over- expression" of DKK-1 was found in: 8/20 of patients with negative serology for both infections (40%; p<0.001), 7/18 of patients with positive anti-HCV antibodies (38.89%; p<0.001) and 11/12 of patients with positive anti-schistosomal antibodies (91.66%; p<0.001). There was no statistically significant correlation between DKK-1 expression and HCV infection (p=0.139) but there was significant correlation between the gene expression and schistosomal infection (p<0.001). These data suggest the role of DKK-1 over-expression in HCC development in patients with combined HCV and schistosomal infections and that induction of the Wnt pathway or using DKK-1 antagonist may represent a key advance in the area of genetic prevention of HCC in these "high-risk" patients.


The protective effect of a novel compound RU 18492, on galactosamine-induced hepatotoxicity in the rat.
A novel compound N-(2-hydroxyethyl)-maleopimarimidyl morpholide (RU 18 492), was investigated for its effect against galactosamine induced hepatitis in rats. It was found to be active in reducing both serum transaminase levels and morphological changes due to the hepatotoxin. The protection was shown to be dose related and a significant reduction in liver damage was seen at doses of 50-400 mg/kg p.o. RU 18 492 was ineffective against carbon tetrachloride induced hepatitis in rats. It was suggested that the protective effect of RU 18 492 was due to its ability to stimulate the smooth endoplasmic reticulum thus inducing a more rapid detoxification of galactosamine.


Early Antibiotic Exposure and Adverse Outcomes in Very Preterm Infants at Low Risk of Early-Onset Sepsis: The EPIPAGE-2 Cohort Study.
To assess the association between early empirical antibiotics and neonatal adverse outcomes in very preterm infants without risk factors for early-onset sepsis (EOS). This is a secondary analysis of the EPIPAGE-2 study, a prospective national population-based cohort that included all liveborn infants at 22-31 completed weeks of gestation in France in 2011. Infants at high risk of EOS (ie, born after preterm labor or preterm premature rupture of membranes or from a mother who had clinical chorioamnionitis or had received antibiotics during the last 72 hours) were excluded. Early antibiotic exposure was defined as antibiotic therapy started at day 0 or day 1 of life, irrespective of the duration and type of antibiotics. We compared treated and untreated patients using inverse probability of treatment weighting based on estimated propensity scores. Among 648 very preterm infants at low risk of EOS, 173 (26.2%) had received early antibiotic treatment. Early antibiotic exposure was not associated with death or late-onset sepsis or necrotizing enterocolitis (OR, 1.04; 95% CI, 0.72-1.50); however, it was associated with higher odds of severe cerebral lesions (OR, 2.71; 95% CI, 1.25-5.86) and moderate-severe bronchopulmonary dysplasia (BPD) (OR, 2.30; 95% CI, 1.21-4.38). Early empirical antibiotic therapy administrated in very preterm infants at low risk of EOS was associated with a higher risk of severe cerebral lesions and moderate-severe BPD.


Fast regulation of axonal growth cone motility by electrical activity.
Axonal growth cones are responsible for the correct guidance of developing axons and the establishment of functional neural networks. They are highly motile because of fast and continuous rearrangements of their actin-rich cytoskeleton. Here we have used live imaging of axonal growth cones of hippocampal neurons in culture and quantified their motility with a temporal resolution of 2 s. Using novel methods of analysis of growth cone dynamics, we show that transient activation of kainate receptors by bath-applied kainate induced a fast and reversible growth cone stalling. This effect depends on electrical activity and can be mimicked by the transient discharge of action potentials elicited in the neuron by intracellular current injections at the somatic level through a patch pipette. Growth cone stalling induced by electrical stimulation is mediated by calcium entry from the extracellular medium as well as by calcium release from intracellular stores that define spatially restricted microdomains directly affecting cytoskeletal dynamics. We propose that growth cone motility is dynamically controlled by transient bursts of spontaneous electrical activity, which constitutes a prominent feature of developing neural networks in vivo.


Structural and functional basis of the serine protease-like hepatocyte growth factor beta-chain in Met binding and signaling.
Hepatocyte growth factor (HGF), a plasminogen-related growth factor, is the ligand for Met, a receptor tyrosine kinase implicated in development, tissue regeneration, and invasive tumor growth. HGF acquires signaling activity only upon proteolytic cleavage of single-chain HGF into its alpha/beta heterodimer, similar to zymogen activation of structurally related serine proteases. Although both chains are required for activation, only the alpha-chain binds Met with high affinity. Recently, we reported that the protease-like HGF beta-chain binds to Met with low affinity (Stamos, J., Lazarus, R. A., Yao, X., Kirchhofer, D., and Wiesmann, C. (2004) EMBO J. 23, 2325-2335). Here we demonstrate that the zymogen-like form of HGF beta also binds Met, albeit with 14-fold lower affinity than the protease-like form, suggesting optimal interactions result from conformational changes upon cleavage of the single-chain form. Extensive mutagenesis of the HGF beta region corresponding to the active site and activation domain of serine proteases showed that 17 of the 38 purified two-chain HGF mutants resulted in impaired cell migration or Met phosphorylation but no loss in Met binding. However, reduced biological activities were well correlated with reduced Met binding of corresponding mutants of HGF beta itself in assays eliminating dominant alpha-chain binding contributions. Moreover, the crystal structure of HGF beta determined at 2.53 A resolution provides a structural context for the mutagenesis data. The functional Met binding site is centered on the "active site region" including "triad" residues Gln(534) [c57], Asp(578) [c102], and Tyr(673) [c195] and neighboring "activation domain" residues Val(692), Pro(693), Gly(694), Arg(695), and Gly(696) [c214-c219]. Together they define a region that bears remarkable resemblance to substrate processing regions of serine proteases. Models of HGF-dependent Met receptor activation are discussed.


Looking back at the future: why Hillarycare failed.
The current Democratic Party candidates for U.S. president, including Hillary Rodham Clinton, have committed themselves to establishing universal health care that will guarantee access to care in time of need, a basic human right still denied in the United States. This commitment is partly a response to the U.S. population's high levels of dissatisfaction (now at unprecedented levels) with the way health care is funded and organized. The article analyzes why a similar commitment by President Bill Clinton in 1992 failed, and challenges some of the main explanations for that failure put forward by protagonists of the White House health care reform task force (chaired by Hillary Clinton). The author emphasizes that the primary reason for the failure was the lack of political will to confront major players in medical care funding, especially the insurance companies and large employers. He postulates that unless such political will exists and unless the system of funding electoral campaigns undergoes major reform-reducing or eliminating the power of financial and economic lobbies in the political process-the United States will not have universal health care. It is a worrisome sign that these lobbies are financing the campaigns of many of today's presidential candidates.


Inclusion body myositis: clonal expansions of muscle-infiltrating T cells persist over time.
Inclusion body myositis (IBM) is a chronic inflammatory myopathy. The muscle histology is characterized by infiltration of T cells, which invade and apparently destroy muscle fibres. This study was performed to investigate whether predominant clones of muscle-infiltrating T cells are identical in different muscles and whether they persist over time in IBM. By reverse transcriptase-polymerase chain reaction, 25 T-cell receptor (TCR) variable beta (Vbeta) chain families and the complementarity-determining region 3 (CDR3) of the TCR were analysed in two different muscle biopsies of four patients with IBM. In two of the patients, the muscle biopsies were obtained from different muscles at one time point, whereas in two patients, the second biopsy was obtained 9 years after the first biopsy. T cells expressing predominant Vbeta families were analysed for clonality by fragment length analysis of the CDR3. Predominant Vbeta families were analysed by DNA sequencing to identify identical clones. Immunohistochemical staining of Vbeta families was performed to study the distribution of T cells expressing identified predominant Vbeta families. The muscle-infiltrating lymphocytes showed restricted expression of TCR Vbeta families. DNA sequencing proved that clonally expanded T cells were identical in different muscles and persisted 9 years after the first biopsy. Immunohistochemical analysis with Vbeta family-specific antibodies demonstrated the endomysial localization of these T cells in inflammatory cell infiltrates. Our results show that in IBM there is clonal restriction of TCR expression in muscle-infiltrating lymphocytes. Identical T-cell clones predominate in different muscles, and these clones persist for many years. These results indicate an important, continuous, antigen-driven inflammatory reaction in IBM.


New light on an old debate: does the RCN-PtCl2 bond include any back-donation? RCN←PtCl2 backbonding vs. the IR νC[triple bond, length as m-dash]N blue-shift dichotomy in organonitriles-platinum(ii) complexes. A thorough density functional theory - energy decomposition analysis study.
For a series of organonitrile [RCN (R = Me, CF3, Ph, CH3Ph, CF3Ph)] ligands, the nature of the N-Pt bond in the related cis-/trans-(RCN)2PtCl2 complexes has been computationally investigated by Density Functional Theory. A fragment based bond analysis has been performed in the canonical Kohn-Sham molecular orbitals framework, and it has been ultimately assessed that this bond is characterized both by N→Pt σ and by N←Pt π contributions. Voronoi Deformation Density charges further confirms the occurrence of N←Pt π interactions. Moreover, the Energy Decomposition Analysis-Natural Orbital for Chemical Valence (EDA-NOCV) method shows that the strength of the N←Pt π interaction is not negligible by contributing to about 30-40% of the total orbital interaction. Finally, the well-known νC[triple bond, length as m-dash]N blue-shift occurring upon coordination to PtII, has been thoroughly investigated by exploiting the EDA-NOCV and by evaluating νC[triple bond, length as m-dash]N and force constants. The origin of the νC[triple bond, length as m-dash]N blue-shift in these systems has been discussed on the basis of the CN bond polarization. N←Pt π backbonding causes only a systematic decrease of the observed νC[triple bond, length as m-dash]N blue-shift when compared to the one calculated for RCN-X (X = H+, alkaline, Lewis acids) herein reported (X = purely σ acceptors).


Treatment options for hyponatremia in heart failure.
Hyponatremia is independently associated with adverse outcomes in patients with congestive heart failure (CHF). The primary cause of hyponatremia in CHF is the inappropriate secretion of the antidiuretic hormone, arginine vasopressin (AVP). The binding of AVP to V(2) receptors in the renal collecting duct promotes water retention, a process that can lead to dilutional hyponatremia as well as increased ventricular preload. AVP could also exacerbate the course of CHF by interacting with V(1A) receptors on vascular smooth muscle cells and myocytes. Conventional treatment of hyponatremia in CHF is based largely on water restriction, which is neither effective nor well tolerated. Current research is exploring V(2)- and dual V(1A)/V(2)-receptor antagonism for the treatment of hyponatremia, as well as for the congestion and edema associated with CHF, since AVP-receptor antagonists may offer benefits in comparison to conventional loop diuretics. Clinical trials in patients with hyponatremia and CHF using both selective and nonselective vasopressin antagonists have demonstrated the effectiveness of these agents in correcting this common electrolyte abnormality.


Optimization of the Conical Angle Design in Conical Implant-Abutment Connections: A Pilot Study Based on the Finite Element Method.
Conical implant-abutment connections are popular for their excellent connection stability, which is attributable to frictional resistance in the connection. However, conical angles, the inherent design parameter of conical connections, exert opposing effects on 2 influencing factors of the connection stability: frictional resistance and abutment rigidity. This pilot study employed an optimization approach through the finite element method to obtain an optimal conical angle for the highest connection stability in an Ankylos-based conical connection system. A nonlinear 3-dimensional finite element parametric model was developed according to the geometry of the Ankylos system (conical half angle = 5.7°) by using the ANSYS 11.0 software. Optimization algorithms were conducted to obtain the optimal conical half angle and achieve the minimal value of maximum von Mises stress in the abutment, which represents the highest connection stability. The optimal conical half angle obtained was 10.1°. Compared with the original design (5.7°), the optimal design demonstrated an increased rigidity of abutment (36.4%) and implant (25.5%), a decreased microgap at the implant-abutment interface (62.3%), a decreased contact pressure (37.9%) with a more uniform stress distribution in the connection, and a decreased stress in the cortical bone (4.5%). In conclusion, the methodology of design optimization to determine the optimal conical angle of the Ankylos-based system is feasible. Because of the heterogeneity of different systems, more studies should be conducted to define the optimal conical angle in various conical connection designs.


Randomized, open-label phase 2 study comparing frontline dovitinib versus sorafenib in patients with advanced hepatocellular carcinoma.
Angiogenesis inhibition by the vascular endothelial growth factor receptor (VEGFR) and platelet-derived growth factor receptor (PDGFR) inhibitor sorafenib provides survival benefit in hepatocellular carcinoma (HCC); however, angiogenic escape from sorafenib may occur due to angiogenesis-associated fibroblast growth factor receptor (FGFR) pathway activation. In addition to VEGFR and PDGFR, dovitinib inhibits FGFR. Frontline oral dovitinib (500 mg/day, 5 days on, 2 days off; n = 82) versus sorafenib (400 mg twice daily; n = 83) was evaluated in an open-label, randomized phase 2 study of Asian-Pacific patients with advanced HCC. The primary and key secondary endpoints were overall survival (OS) and time to tumor progression (TTP) as determined by a local investigator, respectively. Patients included in the study were ineligible for surgical and/or locoregional therapies or had disease progression after receiving these therapies. The median OS (95% confidence interval [CI]) was 8.0 (6.6-9.1) months for dovitinib and 8.4 (5.4-11.3) months for sorafenib. The median TTP (95% CI) per investigator assessment was 4.1 (2.8-4.2) months and 4.1 (2.8-4.3) months for dovitinib and sorafenib, respectively. Common any-cause adverse events included diarrhea (62%), decreased appetite (43%), nausea (41%), vomiting (41%), fatigue (35%), rash (34%), and pyrexia (30%) for dovitinib and palmar-plantar erythrodysesthesia syndrome (66%) and decreased appetite (31%) for sorafenib. Subgroup analysis revealed a significantly higher median OS for patients in the dovitinib arm who had baseline plasma soluble VEGFR1 (sVEGFR1) and hepatocyte growth factor (HGF) below median levels versus at or above the median levels (median OS [95% CI]: sVEGFR1, 11.2 [9.0-13.8] and 5.7 [4.3-7.0] months, respectively [P = .0002]; HGF, 11.2 [8.9-13.8] and 5.9 [5.0-7.6] months, respectively [P = 0.0002]). Dovitinib was well tolerated, but activity was not greater than sorafenib as a frontline systemic therapy for HCC. Based on these data, no subsequent phase 3 study has been planned. (Hepatology 2016;64:774-784).


A chimeric streptokinase with unexpected fibrinolytic selectivity.
Chimeric 59D8-SK was designed to confer fibrin-selectivity to streptokinase by fusion of the Fab fragment of anti-fibrin antibody 59D8 to the N-terminus of streptokinase (SK: Ile1-Lys414). It was expressed in a mouse hybridoma cell line and purified by affinity chromatography on a 59D8-antigen column. Chimeric 59D8-SK is a disulfide-linked heterodimer composed of an antibody light chain (Mr 27,000) and a N-glycosylated chimeric heavy chain (M(r) 90,000). The fibrin targeting by 59D8 increased plasma clot lysis by 2-fold, but connecting 59D8 to SK has provided 59D8-SK several unique properties: (i) 59D8-SK activated human Glu-plasminogen with a significant lag period that coincided with limited proteolysis of 59D8-SK similar to that observed for wild-type SK. In a kinetic study, both gave very similar kinetic parameters for the activation of Glu-plasminogen even though 59D8-SK was N-glycosylated in its SK portion; (ii) 59D8-SK was relatively inactive in human plasma, compared to SK, but it became activated in the presence of clots; (iii) 59D8-SK lysed clots slowly but completely whereas SK lysed clots rapidly but incompletely. Even though the mechanism behind these new properties is not fully understood, they are characteristics of a second-generation plasminogen activator.


Rapamycin reverses elevated mTORC1 signaling in lamin A/C-deficient mice, rescues cardiac and skeletal muscle function, and extends survival.
Mutations in LMNA, the gene that encodes A-type lamins, cause multiple diseases including dystrophies of the skeletal muscle and fat, dilated cardiomyopathy, and progeria-like syndromes (collectively termed laminopathies). Reduced A-type lamin function, however, is most commonly associated with skeletal muscle dystrophy and dilated cardiomyopathy rather than lipodystrophy or progeria. The mechanisms underlying these diseases are only beginning to be unraveled. We report that mice deficient in Lmna, which corresponds to the human gene LMNA, have enhanced mTORC1 (mammalian target of rapamycin complex 1) signaling specifically in tissues linked to pathology, namely, cardiac and skeletal muscle. Pharmacologic reversal of elevated mTORC1 signaling by rapamycin improves cardiac and skeletal muscle function and enhances survival in mice lacking A-type lamins. At the cellular level, rapamycin decreases the number of myocytes with abnormal desmin accumulation and decreases the amount of desmin in both muscle and cardiac tissue of Lmna(-/-) mice. In addition, inhibition of mTORC1 signaling with rapamycin improves defective autophagic-mediated degradation in Lmna(-/-) mice. Together, these findings point to aberrant mTORC1 signaling as a mechanistic component of laminopathies associated with reduced A-type lamin function and offer a potential therapeutic approach, namely, the use of rapamycin-related mTORC1 inhibitors.


[X-ray endovascular surgery in the trauma and orthopedic unit].
The review of X-ray endovascular surgical methods applied in the practice of FSF CITO named after N.N. Priorov since 1970 is presented. The complex evaluation of the altered conditions of performing angiographic examinations for the time present was carried out. The main regulations of anesthesiological provisions for angiographic studies in conditions of an in-patient trauma department are described. The methods of interventional X-ray surgery became use in patients with vertebral column and spinal medulla injuries, traumas with the threat of pulmonary thromboembolism and in patients with bone pathology. More than 200 patients with a traumatic disease of the spinal medulla and scoliotic deformities were investigated. In all patients spinal angiography with the evaluation of the spinal cord blood supply was performed. The results of more than 450 angiographic examinations the patients with bone pathology are presented. In 83 cases X-ray endovascular embolizations of the regional blood flow were done. 63 Cava Filters were implanted in patients with floating pelvic and leg vein thrombosis. Own expertise of the use of tentative Cava Filters is described. These and other data testify of X-ray endovascular methods prove to be in demand in traumatological and orthopedic practice. Their application will open new possibilities for traumatologists and orthopedists in the process of preventive measures, diagnostics and treatment the patients


Leu85 in the beta1-beta2 linker of ASIC1 slows activation and decreases the apparent proton affinity by stabilizing a closed conformation.
Acid-sensing ion channels (ASICs) are proton-activated channels expressed in neurons of the central and peripheral nervous systems where they modulate neuronal activity in response to external increases in proton concentration. The size of ASIC1 currents evoked by a given local acidification is determined by the number of channels in the plasma membrane and by the apparent proton affinities for activation and steady-state desensitization of the channel. Thus, the magnitude of the pH drop and the value of the baseline pH both are functionally important. Recent characterization of ASIC1s from an increasing number of species has made evident that proton affinities of these channels vary across vertebrates. We found that in species with high baseline plasma pH, e.g. frog, shark, and fish, ASIC1 has high proton affinity compared with the mammalian channel. The beta1-beta2 linker in the extracellular domain, specifically by the substitution M85L, determines the interspecies differences in proton affinities and also the time course of ASIC1 macroscopic currents. The mechanism underlying these observations is a delay in channel opening after application of protons, most likely by stabilizing a closed conformation that decreases the apparent affinity to protons and also slows the rise and decay phases of the current. Together, the results suggest evolutionary adaptation of ASIC1 to match the value of the species-specific plasma pH. At the molecular level, adaptation is achieved by substitutions of nonionizable residues rather than by modification of the channel proton sensor.


Positive selection for unpreferred codon usage in eukaryotic genomes.
Natural selection has traditionally been understood as a force responsible for pushing genes to states of higher translational efficiency, whereas lower translational efficiency has been explained by neutral mutation and genetic drift. We looked for evidence of directional selection resulting in increased unpreferred codon usage (and presumably reduced translational efficiency) in three divergent clusters of eukaryotic genomes using a simple optimal-codon-based metric (Kp/Ku). Here we show that for some genes natural selection is indeed responsible for causing accelerated unpreferred codon substitution, and document the scope of this selection. In Cryptococcus and to a lesser extent Drosophila, we find many genes showing a statistically significant signal of selection for unpreferred codon usage in one or more lineages. We did not find evidence for this type of selection in Saccharomyces. The signal of positive selection observed from unpreferred synonymous codon substitutions is coincident in Cryptococcus and Drosophila with the distribution of upstream open reading frames (uORFs), another genic feature known to reduce translational efficiency. Functional enrichment analysis of genes exhibiting low Kp/Ku ratios reveals that genes in regulatory roles are particularly subject to this type of selection. Through genome-wide scans, we find recent selection for unpreferred codon usage at approximately 1% of genetic loci in a Cryptococcus and several genes in Drosophila. Unpreferred codons can impede translation efficiency, and we find that genes with translation-impeding uORFs are enriched for this selection signal. We find that regulatory genes are particularly likely to be subject to selection for unpreferred codon usage. Given that expression noise can propagate through regulatory cascades, and that low translational efficiency can reduce expression noise, this finding supports the hypothesis that translational efficiency may be suppressed in some cases to reduce stochastic noise in gene expression.


A modified strategy for sequence specific assignment of protein NMR spectra based on amino acid type selective experiments.
The determination of the three-dimensional structure of a protein or the study of protein-ligand interactions requires the assignment of all relevant nuclei as an initial step. This is nowadays almost exclusively performed using triple-resonance experiments. The conventional strategy utilizes one or more pairs of three dimensional spectra to obtain redundant information and thus reliable assignments. Here, a modified strategy for obtaining sequence specific assignments based on two dimensional amino acid type selective triple-resonance experiments is proposed. These experiments can be recorded with good resolution in a relatively short time. They provide very specific and redundant information, in particular on sequential connectivities, that drastically increases the ease and reliability of the assignment procedure, done either manually or in an automated fashion. The new strategy is demonstrated with the protein domain PB1 from yeast CDC24p.


Condensing-enriched magnetic photonic barcodes on superhydrophobic surface for ultrasensitive multiple detection.
The ultrasensitive detection of multiple nucleic acids has great significance in a broad range of medical fields since nucleic acid samples are usually of small volumes and highly diluted. Herein, we present a novel magnetic photonic crystal (PhC) barcode-integrated condensing-enriched superhydrophobic platform for the ultrasensitive multiple miRNA detection. Droplets containing targets could suck the hydrogel PhC barcodes without losing their targets due to the difference in wettability between the hydrophilic PhC barcodes and hydrophobic substrate. During the evaporation of water from the droplet microreactors, these targets could be effectively enriched in the barcodes for achieving higher sensitivities. As the encoding signals of the PhC barcodes are the characteristic reflection peaks generated by their ordered physical nanostructures, the barcodes are stable and free from any fluorescent background and photobleaching; thus, they are accurate for distinguishing different targets. In addition, with the integration of magnetic nanoparticles into the hydrogel PhC barcodes, they could be imparted with the features of immobilization during change of medium and flexible movement for controlling the reaction, both of which could facilitate the detection processes. Based on this platform, we have demonstrated that the detection limit of multiple miRNAs is improved by about three orders. Thus, our platform is an ideal alternative for the ultrasensitive simultaneous multiplex analysis in medical fields.


Depresión en el adulto mayor intervenido quirúrgicamente.
To determine the degree of depression in elderly adults after surgery and its relation with the duration of anesthesia. We conducted an observational, comparative, prospective and longitudinal study. We included 73 elderly adults aged 60 scheduled for different surgical procedures. Their degree of depression was evaluated prior to and after the surgery with the short version of the Yasavage Geriatric Depression Scale. They were classified according to the score: no depression (0-5), mild depression (6-9) and established depression (10-15). The relation of depression with anesthesia duration was determined. The sample size was calculated for proportions. Descriptive statistics were used as well as χ2 (p < 0.05). In the first evaluation 47 patients (64%) were not depressed, 21 (29%) had mild depression and 5 (7%) had established depression. In the second evaluation, we found that 44 patients (60%) were not depressed, 21 (29%) had mild depression and 8 (11%) had established depression. The relation between depression and anesthesia duration was χ2 = 0.81. We did not establish a relation between depression and anesthesia duration in surgically intervened elderly adults.


[Thrombosis in ovarian hyperstimulation syndrome].
Ovarian hyperstimulation syndrome is a rare complication of IVF treatment. In severe ovarian hyperstimulation syndrome there is an increased risk of thrombo-embolic events. Venous thrombotic complications tend to occur several weeks after the resolution of ovarian hyperstimulation syndrome. Predominant sites of venous thrombosis in OHSS are the upper extremities. A 34-year old woman presented to the emergency department with complaints about a painful swelling in the neck and a bluish discoloration of the right arm. She had been admitted to the hospital six weeks before, because of shortness of breath from pleural effusion due to ovarian hyperstimulation syndrome. The diagnosis of jugular vein and subclavian vein thrombosis was made. She was treated with therapeutic doses of low-molecular-weight heparin. Venous thrombosis is a rare but serious complication of ovarian hyperstimulation syndrome. For patients who require admission because of severe ovarian hyperstimulation syndrome, thromboprophylaxis should be continued several weeks after discharge.


Th17 Cells, Glucocorticoid Resistance, and Depression.
Depression is a severe mental disorder that disrupts mood and social behavior and is one of the most common neuropsychological symptoms of other somatic diseases. During the study of the disease, a number of theories were put forward (monoamine, inflammatory, vascular theories, etc.), but none of those theories fully explain the pathogenesis of the disease. Steroid resistance is a characteristic feature of depression and can affect not only brain cells but also immune cells. T-helper cells 17 type (Th17) are known for their resistance to the inhibitory effects of glucocorticoids. Unlike the inhibitory effect on other subpopulations of T-helper cells, glucocorticoids can enhance the differentiation of Th17 lymphocytes, their migration to the inflammation, and the production of IL-17A, IL-21, and IL-23 in GC-resistant disease. According to the latest data, in depression, especially the treatment-resistant type, the number of Th17 cells in the blood and the production of IL-17A is increased, which correlates with the severity of the disease. However, there is still a significant gap in knowledge regarding the exact mechanisms by which Th17 cells can influence neuroinflammation in depression. In this review, we discuss the mutual effect of glucocorticoid resistance and Th17 lymphocytes on the pathogenesis of depression.


Chebulic Acid Prevents Hypoxia Insult via Nrf2/ARE Pathway in Ischemic Stroke.
Excessive reactive oxygen species (ROS) production contributes to brain ischemia/reperfusion (I/R) injury through many mechanisms including inflammation, apoptosis, and cellular necrosis. Chebulic acid (CA) isolated from Terminalia chebula has been found to have various biological effects, such as antioxidants. In this study, we investigated the mechanism of the anti-hypoxic neuroprotective effect of CA in vitro and in vivo. The results showed that CA could protect against oxygen-glucose deprivation/reoxygenation (OGD/R) induced neurotoxicity in SH-SY5Y cells, as evidenced by the enhancement of cell viability and improvement of total superoxide dismutase (T-SOD) in SH-SY5Y cells. CA also attenuated OGD/R-induced elevations of malondialdehyde (MDA) and ROS in SH-SY5Y cells. Nuclear factor-E2-related factor 2 (Nrf2) is one of the key regulators of endogenous antioxidant defense. CA acted as antioxidants indirectly by upregulating antioxidant-responsive-element (ARE) and Nrf2 nuclear translocation to relieve OGD/R-induced oxidative damage. Furthermore, the results showed that CA treatment resulted in a significant decrease in ischemic infarct volume and improved performance in the motor ability of mice 24 h after stroke. This study provides a new niche targeting drug to oppose ischemic stroke and reveals the promising potential of CA for the control of ischemic stroke in humans.


Northeastern Conference on Avian Diseases from 1928-2021: 93 Years of Contributions to Organized Avian Medicine.
Bacillary white diarrhea in chickens was a major disease concern for the poultry industry during the early 1900s. Drs. L. F. Rettger and W. R. Hinshaw organized a meeting in 1928 to discuss methods for controlling this disease. In this meeting, representatives of five northeastern states discussed approaches to test for the presence of the etiological agent of bacillary white diarrhea, namely, Salmonella enterica subsp. enterica serovar Pullorum. Meeting attendees decided to have a yearly meeting of the Northeastern Conference of Laboratory Workers in Bacillary White Diarrhea. The next year, the name was changed to Conference of Laboratory Workers in Pullorum Disease Eradication, which was changed to Northeastern Conference on Avian Diseases (NECAD) in 1957. Not only has NECAD been important for the control of pullorum disease but also, starting with the fifth Annual Conference in 1932, other poultry diseases became an official part of the program. As such, NECAD served for a long time as the premier organization to present new information on avian diseases. The success of NECAD was based on the work of the many committees, which are described in detail in this review. For example, the antigen committee started officially in 1929 and remained active until at least 1987. The main task of this committee was to evaluate Salmonella Pullorum strains to be used by all participants in the pullorum antibody testing programs. NECAD started as a closed organization with participants from universities and government organizations but did not allow representatives from commercial groups until 1968 when all American Association of Avian Pathologists (AAAP) members in the Northeastern United States could participate. The journal Avian Diseases started with discussions by Dr. P. P. Levine with M. S. Cover, H. L. Chute, R. F. Gentry, E. Jungherr, and H. Van Roekel about the idea that NECAD would sponsor a journal dealing specifically with avian diseases. During the first few years of Avian Diseases publication, many articles including abstracts came from the NECAD Annual Conferences. The importance of NECAD as a precursor for other regional meetings and the AAAP and as a forum for graduate students to present their research are described. Several recipients of the award for the best paper presented by a graduate student have continued to work in avian disease research. The decline in the participation of scientists in the late 1990s and early 2000s was discussed extensively in 2006 and led to a merger of the NECAD meeting with the Pennsylvania Poultry Sales and Service Conference. Due to the COVID-19 pandemic, the 92nd Annual Conference was a virtual meeting in 2020. Fortunately, the 93rd Annual Conference in 2021 was an in-person meeting held in State College, PA.


Imaging system for nerve and fiber tract morphometry: components, approaches, performance, and results.
This report describes the imaging system that was developed to recognize, count, size, and evaluate shapes of transverse myelinated fiber (MF) profiles in nerves and fiber tracts automatically and by operator interaction. Automatic analysis, without operator interaction, will either miss a variable percentage of small MFs (using thresholds that discriminate against all other tissue profiles) or include most MFs but spuriously detect other tissue profiles (using more sensitive thresholds). Systems such as this must therefore be operator-interactive. The system that was developed will, in a favorable histologic section, automatically detect and border myelin in more than 85% of MFs without inclusion of other tissue elements. The remainder of the MFs are then identified by the operator with the digitizer pen, and the myelin is automatically bordered or, in rare cases, drawn in. Very reliable, reproducible, and rapid measurements of MF number, size, and shape can be obtained with this system. From evaluation of the size and shape of MFs in semithin sections in the light microscope as compared with the same fibers in adjacent thin sections in the electron microscope, measurements by this system can be expressed as if they were obtained on thin sections. Evaluations can be analyzed statistically, with results printed out, displayed on a video screen, and graphed. Such a system will be useful in evaluating and following morphologic changes in number, size, and shape of MFs in development, aging, regeneration, neurotoxicity, and various diseases.


Unusual structural features and assembly of gap and pleated septate junctions in embryonic cockroach CNS.
Junctional assembly in the developing CNS in cockroach embryos has been studied during the last half of neurogenesis. Atypical linear tracts of gap junctions are found to develop between attenuated cytoplasmic glial cell processes and their overlying perineurial cells during the last third of development. During both perineurial and glial gap-junctional formation, 13 nm E face (EF) intramembrane particles (IMPs), such as are characteristic of arthropod gap junctions, are seen initially as free IMPs; these then become arranged in loose irregular clusters or alignments and finally are aggregated in plaques. P face ridges (or EF grooves), typical of tight junctions, are found on the same perineurial membrane face as assembling gap-junctional PF pits (or EF particles). Pleated separate junctions also develop between adjacent perineurial processes during the last third of embryogenesis; these form by the apparent migration of individual 8 nm PF IMPs into meandering rows, which then become aligned in numerous orderly parallel stacks. Although all these junctions occur on the same perineurial membrane face, the IMPs that form the different junctional types never appear to be confused during junctional assembly. The cues to signal the advent of these precise patterns, however, are unknown.


Actions of angiotensin II on the brain: mechanisms and physiologic role.
Angiotensin II acts on the brain to produce a variety of effects including elevation of arterial blood pressure, increased release of vasopressin and ACTH, stimulation of drinking and sodium appetite, and natriuresis. Many, and possibly all, of these effects can be produced by centrally administered angiotensin II or by circulating angiotensin II, which appear to act at common receptor sites located in the circumventricular organs. Whether these effects are normally produced by blood-borne angiotensin II formed by the renal renin-angiotensin system, by angiotensin II formed centrally by the putative brain renin-angiotensin system, or by both, remains to be determined. A large body of information concerning the site and mechanism of these different central actions of angiotensin II is available, and the physiologic significance of these actions is beginning to be understood. Nevertheless, much additional research will be required before the actions of angiotensin II on the brain are completely understood.


Phenotypic analysis of Neurospora crassa gene deletion strains.
Phenotypic analysis of Neurospora crassa knockout (KO) mutants was used as a vehicle to introduce students to laboratory research. The availability of gene deletion strains was the impetus for the development of a program designed to introduce beginning science students to basic microbiology, genetics, microscopy and beginning bioinformatics. The goal was to provide a research experience, acquire laboratory skills and phenotype hundreds of KO mutants. The data provided by the students was used to build a phenotype database at the Broad Institute at Harvard/MIT for the fungal scientific community. Each mutant analysis consists of five assays that examine growth and morphology, asexual and sexual development using wild-type (parental) strains as a reference. This information indicates how loss of each gene impacts these basic and important processes.


[The yin and the yang of autophagy in cancer cells].
Autophagy is a self-cannibalism process essential for tissue homeostasis, which can be activated following different environmental stressful conditions. In normal cells, autophagy could act as a brake to prevent tumorigenesis, but cancer cells are able to hijack this process to their own benefit, to promote tumor growth and/or tumor resistance to anti-cancer therapies. Scientists and clinicians attempt to modulate this process to improve therapies, using autophagy inhibitors or activators, some of them being tested currently in clinical trials against several types of tumors. Thus, it appears that autophagy is at the center of a showdown between cancer cells and anti-cancer therapies. In this review, we focus on the mechanisms by which autophagy could be either the yin or the yang of cancers.


The effect of dose and cytochrome P450 induction on the metabolism and disposition of the food-borne carcinogen 2-amino-3,8-dimethylimidazo[4,5-f] quinoxaline (MeIQx) in the rat.
The effect of dose and cytochrome P450 induction on the metabolism and disposition of the food-borne carcinogen 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) was investigated in the male Sprague-Dawley rat. Animals were given MeIQx by gavage at doses of 0.01, 0.2 or 20 mg/kg body wt. The phase II conjugates, MeIQx-N2 sulfamate and MeIQx-N2 glucuronide were the predominant metabolites found in urine of non-induced animals at the highest dose treatment. Animals induced with polychlorinated-biphenyl (PCB) produced greater amounts of metabolites hydroxylated at the 5 position of MeIQx which were excreted as glucuronide or sulfate conjugates. At the lowest dose studied, the urinary excretion profile was nearly identical for both animal groups and cytochrome P450 induction had little influence on metabolism. In contrast to high dose exposure, where sulfamate formation was a major route of detoxification, N2 glucuronide formation was the most important metabolic pathway for elimination of MeIQx at low doses. Liver microsomes transformed MeIQx to the genotoxic metabolite 2-hydroxyamino-3,8-dimethylimidazo[4,5-f]quinoxaline (HNOH-MeIQx) and N-hydroxylase activity was 20-fold greater in microsomes obtained from PCB-treated animals than in untreated control animals. The increase in N-hydroxylase activity was discerned in vivo through formation of the metastable N-glucuronide conjugate of HNOH-MeIQx (MeIQx-[HO-N]-Gl). This metabolite accounted for approximately 3% of the dose in bile of PCB-treated rats. In contrast, in the non-induced rat, MeIQx-[HO-N]-Gl was preferentially excreted in urine and accounted for approximately 0.2-1% of the total dose. These results demonstrate that the metabolism of MeIQx in the rat is influenced by both dose and cytochrome P450 induction. The absence of intestinal tumors in the non-induced rat may be partially attributed to the low levels of formation and poor biliary excretion of the N-glucuronide conjugate of the genotoxic metabolite HNOH-MeIQx.


Iterative Design of Visual Analytics for a Clinician-in-the-Loop Smart Home.
In order to meet the health needs of the coming "age wave," technology needs to be designed that supports remote health monitoring and assessment. In this study we design clinician in the loop (CIL), a clinician-in-the-loop visual interface, that provides clinicians with patient behavior patterns, derived from smart home data. A total of 60 experienced nurses participated in an iterative design of an interactive graphical interface for remote behavior monitoring. Results of the study indicate that usability of the system improves over multiple iterations of participatory design. In addition, the resulting interface is useful for identifying behavior patterns that are indicative of chronic health conditions and unexpected health events. This technology offers the potential to support self-management and chronic conditions, even for individuals living in remote locations.


Cavity enhanced absorption spectroscopy using a broadband prism cavity and a supercontinuum source.
We report the design and construction of a cavity enhanced absorption spectrometer using broadband Brewster's angle prism retroreflectors and a spatially coherent 500 nm to >1.75 microm supercontinuum excitation source. Using prisms made from fused silica an effective cavity reflectivity of >99.99% at 1.064 microm was achieved. A proof of principle experiment was performed by recording the cavity enhanced absorption spectrum of the weak b-X (1?0) transition of molecular oxygen at 14529 cm-1 and the fifth overtone of the acetylene C-H stretch at 18430 cm(-1). CCD frames were integrated for 150 sec and 30 sec, with 3 frames (each 100 cm(-1) wide) and 1 frame (266 cm(-1) wide) required to observe the O(2) and C(2)H(2) spectra, respectively. A rms noise equivalent absorption (alpha(min)) of 7.21x10(-8) cm(-1) Hz(-1/2) and 1.28x10(-7) cm(-1) Hz(-1/2) with full width half maximum line widths of 0.18 cm(-1) and 0.44 cm(-1) were achieved for the molecular oxygen band and the acetylene overtone, respectively.


Pharmacoepidemiology of antiretroviral drugs in a teaching hospital in Lagos, Nigeria.
Prescribing, adherence, and adverse drug events to HAART in a large antiretroviral programme in Lagos was evaluated. A retrospective 5 year open cohort study. The AIDS Prevention Initiative in Nigeria (APIN) clinic at LUTH is one of the United States Presidential Emergency Plan for AIDS Relief (PEP-FAR) funded centers for HIV relief program in Nigeria Participants The case files of 390 patients on HAART and attending the APIN clinic were reviewed sequel to random selection. Demographics of the patients and pattern of antiretroviral (ARV) combination drugs prescribed were extracted from their case files. The details of the adverse drug events (ADEs) were extracted from drug toxicity forms regularly filled for each patient. A Chi-square test with Yates correction was used to determine the association between adherence and therapeutic outcome. A total of 2944 prescriptions were assessed. Zidovudine + lamivudine + nevirapine (35.87%) and stavudine + lamivudine + nevirapine (35.63%) were the most frequently prescribed combinations. Over 2000 ADEs were reported with cough (13.3%), fever (8.75%) and skin rashes (8.01%) being the most frequently reported. Drug adherence was associated with good therapeutic outcome (χ(2) = 115.60, p<0.0001). Zidovudine + lamivudine + nevirapine was the most frequently prescribed ARV combination. Cough was the most frequently reported ADE. Interventions aimed at rational prescribing of ARV drugs and improving adherence to antiretroviral drugs is essential for good therapeutic outcome in the treatment of HIV infection.


Evaluation of a gene expression microarray-based assay to determine tissue type of origin on a diverse set of 49 malignancies.
The Tissue of Origin Frozen (TOO-FRZ) assay from Pathwork Diagnostics has been cleared by the Food and Drug Administration as a diagnostic study for malignancies of unknown primary. The goal of this study was to evaluate the performance of TOO-FRZ on a diverse collection of malignancies. We collected a diverse set of 49 malignancies. We classified each case into 1 of 4 groups: common morphology from a tissue type included in the TOO-FRZ assay (n=29), uncommon morphology from a tissue type included in the TOO-FRZ assay (n=10), tumor from a tissue type not included in the TOO-FRZ assay (n=3), and malignancies of unknown primary (n=7). We found strong diagnostic performance for common morphologies from tissue types on the TOO-FRZ [overall accuracy=26 of 29 (90%, 95% CI, 73% to 97%)], with perfect performance in all tissue types except gastric (0 of 2) and pancreatic (1 of 2) tissues. There was a significant decline in performance for uncommon morphologies from tissue types included in the TOO-FRZ assay [6 of 10 (60%) cases with an indeterminate result, 1 of 10 (10%) cases with an incorrect prediction, and 3 of 10 (30%) with a correct prediction] and for tumors from tissue types not included in the assay (incorrect prediction in 2 of 3 cases). For the 7 malignancies of unknown primary in our study set, the TOO-FRZ provided a likely clinically useful result in only 2 of 7 cases. These results provide an insight into the strengths and limitations of this molecular assay for the surgical pathologist, and our findings suggest future directions for research in this area.


Effect of beta-cyclodextrin on aroma release and flavor perception.
Binding and release of volatile compounds to and from beta-cyclodextrin were measured in model aqueous systems using static equilibrium headspace and dynamic headspace dilution. Beta-cyclodextrin decreased the static equilibrium headspace for some volatiles (e.g., ethyl octanoate and decanone) due to binding, but dilution studies demonstrated that binding was readily reversible. Dynamic release of hydrophobic volatile compounds was similar to that observed from emulsions. When beta-cyclodextrin was added to fat free yogurt, the release of a commercial lemon flavoring was modified and was similar to release from a regular fat yogurt. Sensory difference testing confirmed the release results. The data demonstrate that beta-cyclodextrin can be used to modify flavor delivery in both model and real systems; the effects in the latter are sensorially significant.


Rationally Designed MicroRNA-Based Genetic Classifiers Target Specific Neurons in the Brain.
Targeting transgene expression to specific cell types in vivo has proven instrumental in characterizing the functional role of defined cell populations. Genetic classifiers, synthetic transgene constructs designed to restrict expression to particular classes of cells, commonly rely on transcriptional promoters to define cellular specificity. However, the large size of many natural promoters complicates their use in viral vectors, an important mode of transgene delivery in the brain and in human gene therapy. Here, we expanded upon an emerging classifier platform, orthogonal to promoter-based strategies, that exploits endogenous microRNA regulation to target gene expression. Such classifiers have been extensively explored in other tissues; however, their use in the nervous system has thus far been limited to targeting gene expression between neurons and supporting cells. Here, we tested the possibility of using combinatory microRNA regulation to specify gene targeting between neuronal subtypes, and successfully targeted inhibitory cells in the neocortex. These classifiers demonstrate the feasibility of designing a new generation of microRNA-based neuron-type- and brain-region-specific gene expression targeting neurotechnologies.


Calibration-free pulse oximetry based on two wavelengths in the infrared - a preliminary study.
The assessment of oxygen saturation in arterial blood by pulse oximetry (SpO₂) is based on the different light absorption spectra for oxygenated and deoxygenated hemoglobin and the analysis of photoplethysmographic (PPG) signals acquired at two wavelengths. Commercial pulse oximeters use two wavelengths in the red and infrared regions which have different pathlengths and the relationship between the PPG-derived parameters and oxygen saturation in arterial blood is determined by means of an empirical calibration. This calibration results in an inherent error, and pulse oximetry thus has an error of about 4%, which is too high for some clinical problems. We present calibration-free pulse oximetry for measurement of SpO₂, based on PPG pulses of two nearby wavelengths in the infrared. By neglecting the difference between the path-lengths of the two nearby wavelengths, SpO₂ can be derived from the PPG parameters with no need for calibration. In the current study we used three laser diodes of wavelengths 780, 785 and 808 nm, with narrow spectral line-width. SaO₂ was calculated by using each pair of PPG signals selected from the three wavelengths. In measurements on healthy subjects, SpO₂ values, obtained by the 780-808 nm wavelength pair were found to be in the normal range. The measurement of SpO₂ by two nearby wavelengths in the infrared with narrow line-width enables the assessment of SpO₂ without calibration.


[Conflict areas in two psychotherapeutic groups of climacteric women].
An important consideration is to specify the areas of conflict that will be worked on during the psychotherapeutic process in order to develop a specific psychological model for a group intervention with menopausal women. To determine the most relevant areas of psychological conflict for women in the menopausal stage by means of a focalization technique. A descriptive, cross-sectional, longitudinal study with non-experimental design was done in 29 patients from the Perinatology National Institute of Mexico (INPer). Two groups were formed (A and B), each one with 17 participants; however, in the first session only 12 women from group A assisted. Seventeen therapy sessions were analyzed. Group formation and the number of sessions were based on research and patients needs. Women were given psychotherapeutic care once a week at the psychology department of INPer, for 90 minutes by session. Psychotherapeutic management during sessions was in charge of psychologist responsible of climacteric program of psychology department; while Gesell chamber observer was a psychologist with groups' specialization. For the statistical analysis frequency measures were used. Eleven areas were identified, which showed a great similarity in both groups. Main ones were: relation of the patient to relatives or extensive family, relation to the partner and relation to the children. This work favors a dynamic therapeutic process with the patients, in addition to being a technique, which provides structure and guidance during menopause.


Monitoring of hemispheric or brainstem functions with neurophysiologic methods during interventional neuroradiology.
Interventional neuroradiology, sometimes requiring full anesthesia, is now performed in high-risk patients. The neurologic state of an anesthetized patient cannot always be sufficiently evaluated during the neuroradiologic examination. In such cases neurophysiological methods provide information about some aspects of the patient's cerebral condition. Three useful neurophysiological methods for monitoring hemispheric or brainstem functions during the neuroradiologic procedure are on-line or Fourier-transformed electroencephalographic (EEG) analysis; the monitoring of somatosensory-evoked potentials; and the monitoring of brainstem acoustic-evoked potentials. All methods can be done in the operating room with small, mobile equipment.


Bilayer Graphene as a Platform for Bosonic Symmetry-Protected Topological States.
Bosonic symmetry protected topological (BSPT) states, the bosonic analogue of topological insulators, have attracted enormous theoretical interest in the last few years. Although BSPT states have been classified by various approaches, there is so far no successful experimental realization of any BSPT state in two or higher dimensions. In this paper, we propose that a two-dimensional BSPT state with U(1)×U(1) symmetry can be realized in bilayer graphene in a magnetic field. Here the two U(1) symmetries represent total spin S^{z} and total charge conservation, respectively. The Coulomb interaction plays a central role in this proposal-it gaps out all the fermions at the boundary, so that only bosonic charge and spin degrees of freedom are gapless and protected at the edge. Based on the above conclusion, we propose that the bulk quantum phase transition between the BSPT and trivial phase, which can be driven by applying both magnetic and electric fields, can become a "bosonic phase transition" with interactions. That is, only bosonic modes close their gap at the transition, which is fundamentally different from all the well-known topological insulator to trivial insulator transitions that occur for free fermion systems. We discuss various experimental consequences of this proposal.


Detection of moving objects using multi-channel kernel fuzzy correlogram based background subtraction.
In this paper, we examine the suitability of correlogram for background subtraction, as a step towards moving object detection. Correlogram captures inter-pixel relationships in a region and is seen to be effective for modeling the dynamic backgrounds. A multi-channel correlogram is proposed using inter-channel and intra-channel correlograms to exploit full color information and the inter-pixel relations on the same color planes and across the planes. We thereafter derive a novel feature, termed multi-channel kernel fuzzy correlogram, composed by applying a fuzzy membership transformation over multi-channel correlogram. Multi-channel kernel fuzzy correlogram maps multi-channel correlogram into a reduced dimensionality space and is less sensitivity to noise. The approach handles multimodal distributions without using multiple models per pixel unlike traditional approaches. The approach does not require ideal background frames for background model initialization and can be initialized with moving objects also. Effectiveness of the proposed method is illustrated on different video sequences.


Linkage Mapping and Comparative Genomics of Red Drum (Sciaenops ocellatus) Using Next-Generation Sequencing.
Developments in next-generation sequencing allow genotyping of thousands of genetic markers across hundreds of individuals in a cost-effective manner. Because of this, it is now possible to rapidly produce dense genetic linkage maps for nonmodel species. Here, we report a dense genetic linkage map for red drum, a marine fish species of considerable economic importance in the southeastern United States and elsewhere. We used a prior microsatellite-based linkage map as a framework and incorporated 1794 haplotyped contigs derived from high-throughput, reduced representation DNA sequencing to produce a linkage map containing 1794 haplotyped restriction-site associated DNA (RAD) contigs, 437 anonymous microsatellites, and 44 expressed sequence-tag-linked microsatellites (EST-SSRs). A total of 274 candidate genes, identified from transcripts from a preliminary hydrocarbon exposure study, were localized to specific chromosomes, using a shared synteny approach. The linkage map will be a useful resource for red drum commercial and restoration aquaculture, and for better understanding and managing populations of red drum in the wild.


Knockdown of MADD and c-FLIP overcomes resistance to TRAIL-induced apoptosis in ovarian cancer cells.
The clinical utility of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in the treatment of established human malignancies is limited by the development of resistance to TRAIL. We hypothesized that knockdown of map-kinase activating death domain containing protein (MADD), a TRAIL-resistance factor, may overcome TRAIL resistance in ovarian cancer cells. MADD expression in resected ovarian cancer specimens and cell lines was quantified with the use of polymerase chain reaction. Sensitivity of ovarian cancer cell lines to TRAIL, with or without MADD knockdown, was assessed. MADD is expressed at relatively higher levels in human malignant ovarian cancer tissues and cell lines, compared with normal ovarian tissues. The cell lines OVCA429 and OVCAR3 were susceptible, and cell lines CAOV-3 and SKOV-3 were resistant to TRAIL. MADD knockdown in CAOV-3 cells, but not in SKOV-3 cells, conferred TRAIL sensitivity. Knockdown of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-inhibitory protein (c-FLIP) in SKOV-3 cells increased spontaneous and TRAIL-induced apoptosis, which was further increased on MADD knockdown. MADD/c-FLIP(L) knockdown can render TRAIL-resistant ovarian cancer cells susceptible to TRAIL.


Thiol switches in redox regulation of chloroplasts: balancing redox state, metabolism and oxidative stress.
In photosynthesizing chloroplasts, rapidly changing energy input, intermediate generation of strong reductants as well as oxidants and multiple participating physicochemical processes and pathways, call for efficient regulation. Coupling redox information to protein function via thiol modifications offers a powerful mechanism to activate, down-regulate and coordinate interdependent processes. Efficient thiol switching of target proteins involves the thiol-disulfide redox regulatory network, which is highly elaborated in chloroplasts. This review addresses the features of this network. Its conditional function depends on specificity of reduction and oxidation reactions and pathways, thiol redox buffering, but also formation of heterogeneous milieus by microdomains, metabolite gradients and macromolecular assemblies. One major player is glutathione. Its synthesis and function is under feedback redox control. The number of thiol-controlled processes and involved thiol switched proteins is steadily increasing, e.g., in tetrapyrrole biosynthesis, plastid transcription and plastid translation. Thus chloroplasts utilize an intricate and versatile redox regulatory network for intraorganellar and retrograde communication.


Increased prevalence of subclinical hypothyroidism in females in mountainous valley of Kashmir.
Iodine-rich diet is necessary for proper thyroid gland function. Subclinical hypothyroidism (SCH) is associated with serious complications. Substantial numbers of patients have risk of SCH getting converted into primary hypothyroidism. The objectives of the present study are to survey dietary iodine intake pattern in ethnic population of Kashmir and to study the prevalence of SCH. A retrospective, cross-sectional referral hospital study was conducted. Sample size comprised of 2550 patients who were referred to Department of Biochemistry, Government Medical college, Srinagar diagnostic laboratory from OPD and IPD of associated SMHS hospital. Assessment of thyroid function over a period of one year from March 2010 to March 2011 in the serum has been performed by electro-chemiluminescence immunoassay method on ECLIA 2010 fully automatic analyzer. Interview cum questionnaire methods were used to record the patient history and dietary iodine intake pattern. Iodine status of these patients was assessed by measuring urinary iodine excretion. Total patients were 2550 comprising of 44.6% males and 56.4% females. Subjects with elevated and normal thyroid stimulating hormone (TSH) levels in the serum were 30.51 and 69.4% respectively. About 550 patients (21.56%) had subclinical hypothyroidism which includes both males and females. Prevalence of SCH was more in females (81.8%) than in males (18.2%). Most of the patients presenting with SCH were in the age group of 20-65 years. The percentage of SCH amongst the study sample patients was 21.56%, which is much higher as compared to other parts of the world. The highest percentage of SCH was found in females (81.8%) as compared to males (18.2%). On the basis of the present study, we suggest that routine screening of selected populations, especially women between 20 and 65 years of age, may be advocated. Further community level awareness programs need to be organized wherein people in mountainous valley of Kashmir are motivated to take salt in iodized form and diet rich in iodine to ensure proper thyroid gland functioning.


NCCN Practice Guidelines for Breast Cancer.
The therapeutic options for patients with noninvasive or invasive breast cancer are complex and varied. In many situations, the patient and physician have the responsibility to jointly explore and ultimately select the most appropriate option from among the available alternatives. With rare exception, the evaluation, treatment, and follow-up recommendations contained within these guidelines were based largely on the results of past and present clinical trials. However, there is not a single clinical situation in which the treatment of breast cancer has been optimized with respect to either maximizing cure or minimizing toxicity and disfigurement. Therefore, patient and physician participation in prospective clinical trials allows patients not only to receive state-of-the-art cancer treatment but also to contribute to the improvement of treatment of future patients.


Multiple GCD genes required for repression of GCN4, a transcriptional activator of amino acid biosynthetic genes in Saccharomyces cerevisiae.
GCN4 encodes a positive regulator of multiple unlinked genes encoding amino acid biosynthetic enzymes in Saccharomyces cerevisiae. Expression of GCN4 is coupled to amino acid availability by a control mechanism involving GCD1 as a negative effector and GCN1, GCN2, and GCN3 as positive effectors of GCN4 expression. We used reversion of a gcn2 gcn3 double mutation to isolate new alleles of GCD1 and mutations in four additional GCD genes which we designate GCD10, GCD11, GCD12, and GCD13. All of the mutations lead to constitutive derepression of HIS4 transcription in the absence of the GCN2+ and GCN3+ alleles. By contrast, the gcd mutations require the wild-type GCN4 allele for their derepressing effect, suggesting that each acts by influencing the level of GCN4 activity in the cell. Consistent with this interpretation, mutations in each GCD gene lead to constitutive derepression of a GCN4::lacZ gene fusion. Thus, at least five gene products are required to maintain the normal repressed level of GCN4 expression in nonstarvation conditions. Interestingly, the gcd mutations are pleiotropic and also affect growth rate in nonstarvation conditions. In addition, certain alleles lead to a loss of M double-stranded RNA required for the killer phenotype. This pleiotropy suggests that the GCD gene products contribute to an essential cellular function, in addition to, or in conjunction with, their role in GCN4 regulation.


Exercise-induced AMPK activation does not interfere with muscle hypertrophy in response to resistance training in men.
As aerobic exercise (AE) may interfere with adaptations to resistance exercise (RE), this study explored acute and chronic responses to consecutive AE (∼45 min cycling) and RE (4 × 7 maximal knee extensions) vs. RE only. Ten men performed acute unilateral AE + RE interspersed by 15 min recovery. The contralateral leg was subjected to RE. This exercise paradigm was then implemented in a 5-wk training program. Protein phosphorylation, gene expression, and glycogen content were assessed in biopsies obtained from the vastus lateralis muscle of both legs immediately before and 3 h after acute RE. Quadriceps muscle size and in vivo torque were measured, and muscle samples were analyzed for citrate synthase activity and glycogen concentration, before and after training. Acute AE reduced glycogen content (32%; P < 0.05) and increased (P < 0.05) phosphorylation of AMPK (1.5-fold) and rpS6 (1.3-fold). Phosphorylation of p70S6K and 4E-BP1 remained unchanged. Myostatin gene expression was downregulated after acute AE + RE but not RE. Muscle size showed greater (P < 0.05) increase after AE + RE (6%) than RE (3%) training. Citrate synthase activity (18%) and endurance performance (22%) increased (P < 0.05) after AE + RE but not RE. While training increased (P < 0.05) in vivo muscle strength in both legs, normalized and concentric torque increased after RE only. Thus AE activates AMPK, reduces glycogen stores, and impairs the progression of concentric force, yet muscle hypertrophic responses to chronic RE training appear not to be compromised.


Distribution of constitutive (COX-1) and inducible (COX-2) cyclooxygenase in postviral human liver cirrhosis: a possible role for COX-2 in the pathogenesis of liver cirrhosis.
Prostaglandins produced by the action of cyclooxygenases (COX) are important mediators of systemic vasodilatation and inflammation in liver cirrhosis. The aim of this study was to investigate the distribution of COX-1 and COX-2 in postviral cirrhosis. The immunohistochemical expression of the constitutive (COX-1) and the inducible (COX-2) isoenzymes was investigated in 15 patients with cirrhosis after hepatitis B and C infection; three normal control livers were also analysed. COX-2 was absent from normal liver but was highly expressed in cirrhosis, mainly in the inflammatory, sinusoidal, vascular endothelial, and biliary epithelial cells. Low amounts of COX-1 were expressed in both normal and cirrhotic livers, exclusively in sinusoidal and vascular endothelial cells, with no differences seen between normal and cirrhotic livers. COX-2 is overexpressed in liver cirrhosis, and possibly contributes to prostaglandin overproduction, which may be a major component of the inflammation and hyperdynamic circulation associated with cirrhosis. Because COX-2 is thought to contribute to tumour development, high COX-2 production could be a contributor to hepatocellular carcinoma development in cirrhosis. The finding of COX-2 and not COX-1 upregulation in cirrhosis could provide a possible new role for selective COX-2 inhibitors in reducing inflammation and minimising the occurrence of hepatocellular carcinoma in patients with cirrhosis.


A 12-month follow-up for neurological complication after subarachnoid anesthesia in a parturient affected by multiple sclerosis.
Multiple sclerosis is common among women of childbearing age. Neuraxial blocks have been administered to them with reluctance because of the hypothetical risk that local anesthetics might be more histotoxic to neural tissue already compromised by multiple sclerosis. In spite of the lack of uniform guidelines on disorders in pregnancy like multiple sclerosis, and of the published data that sometimes contrast each other, experience gained in recent years has indicated that regional anesthesia is safe even in these patients, but there aren't many published cases. We describe the case of a pregnant woman affected by multiple sclerosis in which we administered spinal anesthesia for a cesarean section, and we analyzed the aspects that literature defines as critical points in this group of patients. The results were favorable with regard to the level, intensity and duration of anesthesia. No neurological exacerbations were recognized during the hospital stay, nor during the follow-up that lasted 12 months.


Temporoparietal Junction Hypoactivity during Pain-Related Empathy Processing in Adolescents with Conduct Disorder.
Background: Lack of empathy has been proposed to account for the characteristic behavioral problems exhibited by adolescents with conduct disorder (CD). Hence, the aim of this study was to determine whether adolescents with CD exhibit atypical affective and cognitive neural empathic responses during pain-related empathy processing. Methods: A total of 30 adolescents with a CD diagnosis and 36 without CD symptoms were recruited from out-patient clinics and local middle schools in the same region, respectively. All 66 participants were subjected to functional magnetic resonance imaging (fMRI) while viewing video clips depicting a face with a neutral expression receiving non-painful stimulation (Q-tip touch) or a face with a painful expression receiving painful stimulation (needle penetration) applied to the left or right cheek. Results: The regions associated with affective and cognitive empathy were activated in the HC group during pain-related empathy processing. Compared to HCs, adolescents with CD showed significantly reduced activation in the bilateral temporoparietal junction (TPJ). Conclusions: Adolescents with CD exhibited dampened hemodynamic responses during pain-related empathy processing in the bilateral TPJ, a region associated with cognitive empathy. These findings are consistent with the hypothesis that adolescents with CD may have a cognitive empathy deficiency.


Ambulatory blood pressure normality: results from the PAMELA study.
To determine ambulatory and home blood pressure means and distributions in relation to clinic blood pressure in a general population. We obtained a random sample of 2400 subjects stratified by sex and 10 year age groups to be representative of residents aged 25-64 years of the city of Monza. Participation rate was 69% (1651 subjects). Blood pressure measurements consisted of clinic blood pressure (average of three measurements, sphygmomanometry), home blood pressure (average of morning and evening measurements, semiautomatic device) and ambulatory blood pressure (automatic readings at 20 min intervals, Spacelabs 90207). Clinic blood pressure was obtained both before and after home and ambulatory blood pressures. Data analysis did not include 213 subjects receiving antihypertensive drug treatment and was therefore limited to 1438 participants. In the 1438 subjects, clinic, home and ambulatory blood pressure showed a normal-like distribution, with a taller peak and a narrower base for ambulatory than for home and clinic values. Clinic, home and ambulatory blood pressures were significantly related to each other (P always < 0.001). The means of the two clinic blood pressures obtained on consecutive days were superimposable (127.4 +/- 17.0/82.3 +/- 9.8 and 128.2 +/- 16.5/81.9 +/- 9.9 mmHg) and both were markedly higher than home and 24 h average blood pressures (8.2 mmHg), which were similar to one another. The differences between clinic and home or 24 h average blood pressure were similar in both sexes but increased with increasing age and clinic blood pressure values. The influence of clinic blood pressure values on the clinic-ambulatory or clinic-home blood pressure differences was more important than age. Although higher than the 24 h average value, daytime average blood pressure was also lower than clinic blood pressure. Night-time blood pressure was markedly lower than the daytime value in both sexes and at all ages. Data from a large and unbiased sample of a general population show that home and 24 h or daytime average blood pressures are much lower than clinic blood pressure. The relatively close correlation between blood pressure values measured with the different methods used has allowed calculation of home and ambulatory blood pressure values corresponding to the accepted upper limit of normality of clinic blood pressure (140/90 mmHg). The upper limit of normality for the population was for both home and ambulatory blood pressures in the range 120-130 and 75-81 mmHg for systolic and diastolic values, respectively, with slight differences depending on sex and age. Taking 140/90 mmHg as the upper normal limit of the population is therefore an error that leads to individuals whose home or ambulatory blood pressures are high being considered as normotensive.


Rat pulmonary responses to inhaled nano-TiO₂: effect of primary particle size and agglomeration state.
The exact role of primary nanoparticle (NP) size and their degree of agglomeration in aerosols on the determination of pulmonary effects is still poorly understood. Smaller NP are thought to have greater biological reactivity, but their level of agglomeration in an aerosol may also have an impact on pulmonary response. The aim of this study was to investigate the role of primary NP size and the agglomeration state in aerosols, using well-characterized TiO₂ NP, on their relative pulmonary toxicity, through inflammatory, cytotoxic and oxidative stress effects in Fisher 344 male rats. Three different sizes of TiO₂ NP, i.e., 5, 10-30 or 50 nm, were inhaled as small (SA) (< 100 nm) or large agglomerates (LA) (> 100 nm) at 20 mg/m³ for 6 hours. Compared to the controls, bronchoalveolar lavage fluids (BALF) showed that LA aerosols induced an acute inflammatory response, characterized by a significant increase in the number of neutrophils, while SA aerosols produced significant oxidative stress damages and cytotoxicity. Data also demonstrate that for an agglomeration state smaller than 100 nm, the 5 nm particles caused a significant increase in cytotoxic effects compared to controls (assessed by an increase in LDH activity), while oxidative damage measured by 8-isoprostane concentration was less when compared to 10-30 and 50 nm particles. In both SA and LA aerosols, the 10-30 nm TiO₂ NP size induced the most pronounced pro-inflammatory effects compared to controls. Overall, this study showed that initial NP size and agglomeration state are key determinants of nano-TiO₂ lung inflammatory reaction, cytotoxic and oxidative stress induced effects.


Measuring COVID-19 health literacy: validation of the COVID-19 HL questionnaire in Spain.
The COVID-19 pandemic has highlighted the importance of health literacy to make informed preventive decisions. A specific COVID-19 health literacy questionnaire (CHL-Q) is included in the COVID-19 Snapshot Monitoring WHO initiative to conduct behavioral insights studies related to COVID-19. The objective was to assess the psychometric properties of a Spanish version of the COVID-19 Health Literacy Questionnaire (CHL-Q). Data quality, acceptability, internal consistency, and construct and structural validity were analyzed. A Rasch analysis was also performed. This cross-sectional, observational study was conducted on the Spanish general population after the first wave of the pandemic and after the end of the general lockdown by an online survey agency. 1033 participants (inclusion criteria were being 18 years or older and living in Spain), was extracted from a panel of approximately 982,000 participants. The sampling was stratified matching the Spanish general population in terms of age, gender, and area of residence. The CHL-Q includes 9 items and assesses people's knowledge, motivation and competencies to access, understand, evaluate, and apply information about COVID-19 in order to make informed decisions. CHL-Q index presented a mean of 33.89 (SD = 9.4), and good fit to the Rasch model (χ2(32) = 34.672, p = 0.342, person separation index = 0.77), with ordered thresholds, unidimensionality, item local independence, and no item bias by sex, age or education level. The CHL-Q showed significant different scores by level of education, experience of infection, confusion related to COVID-19 information and adherence to preventive measures. We found a statistically significant correlation between the CHL-Q index and the total number of preventive measures adopted, COVID-19 knowledge, and information seeking behaviour. The Cronbach´s alpha was 0.87 and the item total corrected correlation, 0.49-0.68. The Spanish version of CHL-Q is a short, adequate, and reliable instrument to measure COVID-19 related health literacy in the Spanish general population. Measuring the CHL in the population can be useful to evaluate whether public authorities, media and the medical and scientific community have been able to reach the population to offer the information in the terms they need it.


Role of glycosylation in transport of vesicular stomatitis virus envelope glycoprotein. A new class of mutant defective in glycosylation and transport of G protein.
A temperature-sensitive mutant (ts gamma 1) of the Cocal serotype of vesicular stomatitis virus synthesizes at the permissive temperature (32 degrees C) a glycoprotein G whose size is smaller (Mr 68,000) than the wild-type (Mr 71,000) and that renders the virion thermolabile. At the nonpermissive temperature (39 degrees C), reduced amounts of noninfectious virus-like particles deficient in G protein were produced. The size of the intracellular G protein was further decreased (Mr 64,000) at the nonpermissive temperature. Biochemical studies including sugar labeling, tryptic peptide analysis, and NH2-terminal sequence analysis of the various glycoproteins suggest that at 32 degrees C a G protein containing a single glycosidic moiety is synthesized. The G protein containing only 1 oligosaccharide residue is transported to the cell surface and is incorporated in infectious virus particles. In contrast, the G protein synthesized at 39 degrees C is nonglycosylated and fails to reach the cell surface. These results suggest that glycosylation of G protein is essential for its transport to the cell surface, and the presence of a single carbohydrate chain is sufficient for this purpose.


Effects of statins on endothelium and endothelial progenitor cell recruitment.
Statins appear to be potent drugs with a variety of pleiotropic effects with vasculoprotective and cardioprotective activity. The beneficial effects of statins on endothelial cells as well as on endothelial cell function appear to be related to improved nitric oxide bioavailability. Mechanistically, statins induce endothelial nitric oxide synthase mRNA stability in endothelial cells and promote endothelial nitric oxide synthase activity through a PI3K/Akt dependent pathway, which is a common signal transduction pathway shared by growth factors such as vascular endothelial growth factors or fibroblast growth factors (FGFs), estrogens, or statins. Furthermore, statins have potent antiinflammatory capacities by potently interfering with the generation of reactive oxygen species or activating scavenging systems for free radicals such as the thioredoxin system. These mechanisms might all contribute to improved NO bioavailability and confer the beneficial actions of statins. The proangiogenic properties of statins and their effects on reendothelialization following vessel injury include novel actions such as the mobilization, differentiation, and improved survival of endothelial progenitor cells. Statin therapy might reverse the impaired functional regeneration capacities seen in patients with risk factors for coronary artery disease or documented active coronary artery disease by specifically interacting with progenitor cell function. Accordingly, augmentation of functionally active endothelial progenitor cells with improved homing capacity will be a critical step in advancing therapeutic neovascularization as well as reendothelialization in patients with coronary artery disease.


Omics profiling identifies the regulatory functions of the MAPK/ERK pathway in nephron progenitor metabolism.
Nephron endowment is defined by fetal kidney growth and crucially dictates renal health in adults. Defects in the molecular regulation of nephron progenitors contribute to only a fraction of reduced nephron mass cases, suggesting alternative causative mechanisms. The importance of MAPK/ERK activation in nephron progenitor maintenance has been previously demonstrated, and here, we characterized the metabolic consequences of MAPK/ERK deficiency. Liquid chromatography/mass spectrometry-based metabolomics profiling identified 42 reduced metabolites, of which 26 were supported by in vivo transcriptional changes in MAPK/ERK-deficient nephron progenitors. Among these, mitochondria, ribosome and amino acid metabolism, together with diminished pyruvate and proline metabolism, were the most affected pathways. In vitro cultures of mouse kidneys demonstrated a dosage-specific function for pyruvate in controlling the shape of the ureteric bud tip, a regulatory niche for nephron progenitors. In vivo disruption of proline metabolism caused premature nephron progenitor exhaustion through their accelerated differentiation in pyrroline-5-carboxylate reductases 1 (Pycr1) and 2 (Pycr2) double-knockout kidneys. Pycr1/Pycr2-deficient progenitors showed normal cell survival, indicating no changes in cellular stress. Our results suggest that MAPK/ERK-dependent metabolism functionally participates in nephron progenitor maintenance by monitoring pyruvate and proline biogenesis in developing kidneys.


Assessment of hepatitis C infection in injecting drug users attending an addiction treatment clinic.
Injecting drug users represent a high risk group for hepatitis C (HCV) infection. Currently, screening of this group for HCV is inconsistently implemented. We designed a HCV assessment algorithm and sought to determine the frequency with which injecting drug users completed the assessment process. Prospective study of a HCV assessment algorithm in the setting of a specialist outpatient addiction treatment clinic. Participants consisted of consecutive new attenders over a six-month period with a history of injecting. Only 21 (18%) of 119 patients reached a satisfactory endpoint of assessment. Forty-eight injectors were tested for antibody to HCV, of whom 26 (54%) tested positive. Thirteen of those with positive test results were no longer attending when the test result became available. Only four of the 19 patients who were referred, attended the on-site hepatology clinic. Thorough screening of injecting drug users for HCV within one treatment service is difficult. There is a need for explicit policies on this issue involving co-operation between primary care providers and addiction services and hepatology services.


Toxicity of benzyl paraben on aquatic as well as terrestrial life.
Parabens are derivatives of alkyl esters of p-hydroxybenzoic acid and come in different classes. These compounds are primarily used as antimicrobial preservative agents in many commercial products, including cosmetics and pharmaceuticals. Accordingly, Benzyl paraben (BeP) is known to be a potential endocrine disruptor. The aim of this study was to determine the toxicity of benzyl paraben (BeP) on aquatic and terrestrial organisms, specifically Scenedesmus sp., Moina macrocopa, and Eisenia fetida. All the organisms were treated with different concentrations of BeP (0.025 mg/L and 1000 mg/L), and LC25, LC50, and LC90 values were used to measure the toxicity levels. Results showed the LC values of BeP for M. macrocopa (3.3 mg/L, 4.7 mg/L, 7.3 mg/L) and E. fetida (173.2 mg/L, 479.8 mg/L, 1062 mg/L), respectively. Toxicity tests on green algae (Scenedesmus sp.) were conducted, the green algae were subjected to various BeP concentration. At 50 mg/L of BeP, cell viability was reduced to 56.2% and the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay indicated 35.4% viable cells. The chlorophyll value and the biochemical parameters of the algal cells were corroborative with the cell viability test. Lethal indices (LC50) for M. macrocopa and E. fetida were evaluated for their toxicity on biochemical properties and were found to be catalase (0.111 mg/L, 0.5 mg/L), lipid peroxidation (0.072 mg/L, 0.056 mg/L), and total protein (0.309 mg/L, 0.314 mg/L), respectively. Overall, this study demonstrated the toxic impact of BeP on non-target aquatic as well as terrestrial species.


Provocative discography in volunteer subjects with mild persistent low back pain.
Whether discographic injections would be positive in subjects with benign persistent "backache" who are not seeking treatment is unknown. This information is important, because benign backache undoubtedly co-exists in patients with chronic low back pain (CLBP) illness that is not discogenicin origin. If these subjects had a high rate of positive discography, the high background incidence of common backache would allow many positive tests in patients in whom discogenic processes were unrelated to their severe CLBP illness. Conversely, if subjects with benign low back pain rarely if ever had significant concordant pain reproduction on disc injections, the basic tenet of discographic diagnosis would be strengthened. To compare, using a strict experimental design, the relative pain and concordancy response to provocative discography in subjects with clinically insignificant "backache" and clinical subjects with CLBP illness considering surgical treatment. Comparison of experimental disc injections in subjects with persistent mild backache and those with chronic low back pain (CLBP) illness. Twenty-five subjects with mild persistent low back pain (LBP) were recruited for an experimental discography study. Subjects were recruited from a clinical study of patients having had cervical spine surgery. Inclusion criteria required that subjects not be receiving or seeking medical treatment for LBP, be taking no medications for backache, have no activity restrictions because of LBP, and have normal psychometric scores. To more closely approximate the pain behavior in CLBP illness, 50% (12) of the "backache" group were recruited with a chronic painful condition (neck/shoulder) unrelated to the low back. CLBP subjects, patients coming to discography for consideration of surgical treatment, were used as control subjects. Results of discography were determined using the criteria of Walsh et al.: pain response of 3 or greater, two or more pain behaviors, a negative "control" discographic injection, and a similar or exact concordancy rating. Discography was performed on experimental subjects and control patients. Experienced raters, who were blinded to control versus experimental status of the subjects, scored the magnetic resonance image, discogram, psychometric tests and discography videotapes of the subjects' pain behavior. Thirteen of 25 volunteer subjects had pain rated as "bad" or worse with disc injection. There were 12 painful and fully concordant disc injections in 9 of these 25 "backache" subjects (36%). These injections met all the Walsh et al. criteria for a positive diagnosis of discogenic pain. All positive discs had annular disruption to or through the outer annulus. Of the 9 subjects with positive discograms, 3 had no chronic pain states and 6 did. All subjects with positive injections had negative control discs. In comparison, in 52 subjects with CLBP illness 38 (73%) had at least one positive disc injection. In a group of volunteer subjects with persistent "backache," 36% were found to have significant pain on disc injection, which is reported to be concordant with their usual pain. The presence of positive concordant pain responses and negative control discs in 33% of subjects without CLBP illness seriously challenges the specificity of provocative discography in identifying a clinically relevant spinal pathology.


Activity of hepatic but not skeletal muscle carnitine palmitoyltransferase enzyme is depressed by intravenous glucose infusions in lactating dairy cows.
A positive energy balance in dairy cows pre-partum may decrease hepatic carnitine palmitoyltransferase (CPT) enzyme activity, which might contribute to disturbances of lipid metabolism post-partum. The purpose of this study was to investigate whether skeletal muscle CPT activity can also be downregulated during positive energy balance. Mid-lactating dairy cows were maintained on intravenous infusion of either saline (control) or glucose solutions that increased linearly over 24 days, remained at the 24-day level until day 28 and were suspended thereafter. Liver and skeletal muscle biopsies, as well as four diurnal blood samples, were taken on days 0, 8, 16, 24, and 32, representing infusion levels equivalent to 0%, 10%, 20%, 30% and 0% of the net energy for lactation (NE(L)) requirement respectively. Glucose infusion increased serum insulin concentrations on day 16 and 24 while plasma glucose levels were increased at only a single time point on day 24. Serum beta-hydroxybutyric acid concentrations decreased between day 8 and 24; whereas changes in non-esterified fatty acids were mostly insignificant. Total lipid contents of liver and skeletal muscle were not affected by treatment. Hepatic CPT activity decreased with glucose infusion (by 35% on day 24) and remained decreased on day 32. Hepatic expression levels of CPT-1A and CPT-2 mRNA were not significantly altered but tended to reflect the changes in enzyme activity. In contrast to the liver, no effect of glucose infusion was observed on skeletal muscle CPT activity. We conclude that suppression of CPT activity by positive energy balance appears to be specific for the liver in mid-lactating dairy cows.


Treatment Changes among Users of Non-Vitamin K Antagonist Oral Anticoagulants in Atrial Fibrillation.
Patients with atrial fibrillation discontinuing anticoagulant therapy are left unprotected against ischaemic stroke. Further, switching between oral anticoagulants may be associated with a transiently increased risk of bleeding or thromboembolism. However, there is a paucity of real-life data on pattern of switching and discontinuation of oral anticoagulants. To address this, we conducted a nationwide drug utilization study including all registered Danish atrial fibrillation patients initiating a non-VKA oral anticoagulant (NOAC) between August 2011 and February 2016. We assessed changes in anticoagulant treatment, including switching between oral anticoagulants and discontinuation of NOACs, and explored patient characteristics predicting these changes. We identified 50,632 patients with atrial fibrillation initiating NOAC therapy within the study period. The majority initiated dabigatran (49.9%) and one-third had previously used VKA. Within 1 year, 10.1% switched to VKA, 4.8% switched to another NOAC and 14.4% discontinued treatment. The frequencies of switching to VKA and discontinuation were highest among NOAC users of young age (<55 years) and with low CHA2 DS2 -VASc score (=0). However, the majority of patients (87.3%) stopping NOAC treatment had a CHA2 DS2 -VASc score ≥1. We conclude that switching from VKA to NOAC, and to a lesser extent from NOAC to VKA, is common, as is early treatment discontinuation. The majority of treatment changes are observed in patients at increased risk of stroke. More research is warranted on the risks of bleeding and thromboembolism associated with switching and discontinuation of NOACs as well as the underlying reasons why these treatment changes occur.


Structural basis of kainate subtype glutamate receptor desensitization.
Glutamate receptors are ligand-gated tetrameric ion channels that mediate synaptic transmission in the central nervous system. They are instrumental in vertebrate cognition and their dysfunction underlies diverse diseases. In both the resting and desensitized states of AMPA and kainate receptor subtypes, the ion channels are closed, whereas the ligand-binding domains, which are physically coupled to the channels, adopt markedly different conformations. Without an atomic model for the desensitized state, it is not possible to address a central problem in receptor gating: how the resting and desensitized receptor states both display closed ion channels, although they have major differences in the quaternary structure of the ligand-binding domain. Here, by determining the structure of the kainate receptor GluK2 subtype in its desensitized state by cryo-electron microscopy (cryo-EM) at 3.8 Å resolution, we show that desensitization is characterized by the establishment of a ring-like structure in the ligand-binding domain layer of the receptor. Formation of this 'desensitization ring' is mediated by staggered helix contacts between adjacent subunits, which leads to a pseudo-four-fold symmetric arrangement of the ligand-binding domains, illustrating subtle changes in symmetry that are important for the gating mechanism. Disruption of the desensitization ring is probably the key switch that enables restoration of the receptor to its resting state, thereby completing the gating cycle.


An Actor-Critic architecture and simulator for goal-directed Brain-Machine Interfaces.
The Perception-Action Cycle (PAC) is a central component of goal-directed behavior because it links internal percepts with external outcomes in the environment. Using inspiration from the PAC, we are developing a Brain-Machine Interface control architecture that utilizes both motor commands and goal information directly from the brain to navigate to novel targets in an environment. An Actor-Critic algorithm was selected for decoding the neural motor commands because it is a PAC-based computational framework where the perception component is implemented in the critic structure and the actor is responsible for taking actions. We develop in this work a biologically realistic simulator to analyze the performance of the decoder in terms of convergence and target acquisition. Experience from the simulator will guide parameter selection and assist in understanding the architecture before animal experiments. By varying the signal to noise ratio of the neural input and error signal, we were able to demonstrate how the learning rate and initial conditions affect a motor control target selection task. In this framework, the naïve decoder was able to reach targets in the presence of noise in the error signal and neural motor command with 98% accuracy.