The impact of single-nucleotide polymorphisms of human apurinic/apyrimidinic endonuclease 1 on specific DNA binding and catalysis.
Human apurinic/apyrimidinic (AP) endonuclease APE1 is a crucial enzyme of the base excision repair (BER) pathway, which is in charge of recognition and initiation of removal of AP-sites in DNA. It is known that some single-nucleotide polymorphism (SNP) variants of APE1 have a reduced activity as compared to wild-type APE1. It has been hypothesized that genetic variation in APE1 might be responsible for an increased risk of some types of cancer. In the present work, analysis of SNPs of the APE1 gene was performed to select the set of variants having substitutions of amino acid residues on the surface of the enzyme globule and in the DNA-binding site, thereby affecting protein-protein interactions or the catalytic reaction, respectively. For seven APE1 variants (R221C, N222H, R237A, G241R, M270T, R274Q, and P311S), conformational dynamics and catalytic activities were examined. The conformational changes in the molecules of APE1 variants and in a DNA substrate were recorded as fluorescence changes of Trp and 2-aminopurine residues, respectively, using the stopped-flow technique. The results made it possible to determine the kinetic mechanism underlying the interactions of the APE1 variants with DNA substrates, to calculate the rate constants of the elementary stages, and to identify the stages of the process affected by mutation.


Position and length of fatty acids strongly affect receptor selectivity pattern of human pancreatic polypeptide analogues.
Pancreatic polypeptide (PP) is a satiety-inducing gut hormone targeting predominantly the Y4 receptor within the neuropeptide Y multiligand/multireceptor family. Palmitoylated PP-based ligands have already been reported to exert prolonged satiety-inducing effects in animal models. Here, we suggest that other lipidation sites and different fatty acid chain lengths may affect receptor selectivity and metabolic stability. Activity tests revealed significantly enhanced potency of long fatty acid conjugates on all four Y receptors with a preference of position 22 over 30 at Y1 , Y2 and Y5 receptors. Improved Y receptor selectivity was observed for two short fatty acid analogues. Moreover, [K(30)(E-Prop)]hPP2-36 (15) displayed enhanced stability in blood plasma and liver homogenates. Thus, short chain lipidation of hPP at key residue 30 is a promising approach for anti-obesity therapy because of maintained selectivity and a sixfold increased plasma half-life.


Association Between Thyroid Function and Objective and Subjective Sleep Quality in Older Men: The Osteoporotic Fractures in Men (MrOS) Study.
To determine the association between thyroid hormone levels and sleep quality in community-dwelling men. Among 5,994 men aged ≥65 years in the Osteoporotic Fractures in Men (MrOS) study, 682 had baseline thyroid function data, normal free thyroxine (FT4) (0.70 ≤ FT4 ≤ 1.85 ng/dL), actigraphy measurements, and were not using thyroid-related medications. Three categories of thyroid function were defined: subclinical hyperthyroid (thyroid-stimulating hormone [TSH] <0.55 mIU/L), euthyroid (TSH, 0.55 to 4.78 mIU/L), and subclinical hypothyroid (TSH >4.78 mIU/L). Objective (total hours of nighttime sleep [TST], sleep efficiency [SE], wake after sleep onset [WASO], sleep latency [SL], number of long wake episodes [LWEP]) and subjective (TST, Pittsburgh Sleep Quality Index score, Epworth Sleepiness Scale score) sleep quality parameters were measured. The association between TSH and sleep quality was examined using linear regression (continuous sleep outcomes) and log-binomial regression (categorical sleep outcomes). Among the 682 men examined, 15 had subclinical hyperthyroidism and 38 had subclinical hypothyroidism. There was no difference in sleep quality between subclinical hypothyroid and euthyroid men. Compared to euthyroid men, subclinical hyperthyroid men had lower mean actigraphy TST (adjusted mean difference [95% confidence interval (CI)], -27.4 [-63.7 to 8.9] minutes), lower mean SE (-4.5% [-10.3% to 1.3%]), and higher mean WASO (13.5 [-8.0 to 35.0] minutes]), whereas 41% had increased risk of actigraphy-measured TST <6 hours (relative risk [RR], 1.41; 95% CI, 0.83 to 2.39), and 83% had increased risk of SL ≥60 minutes (RR, 1.83; 95% CI, 0.65 to 5.14) (all P>.05). Neither subclinical hypothyroidism nor hyperthyroidism is significantly associated with decreased sleep quality.


Effects of Intraperitoneal Injection of Allogeneic Bone Marrow-derived Mesenchymal Stem Cells on Bronchiolitis Obliterans in Mice Model.
Bone marrow-derived mesenchymal stem cells (BMSCs) can ameliorate a variety of lung diseases such as asthma, lung fibrosis, and acute lung injury by its anti-inflammatory and immunmodulatory effects. In this study, we developed a mouse model of bronchiolitis obliterans (BO) and evaluated the effects of the intraperitoneal administration of BMSCs on lung histopathology and cytokine levels. 25 BALB/c mice were divided into four groups; control group (Group I), BO developed and 1x106 BMSCs-injected group (Group II), non-BO, 1x106 BMSCs-injected group (Group III), and BO developed and saline-injected group (Group IV). Histological and immunohistochemical findings of the lung tissue and the migration of BMSCs to the lung were evaluated using light and confocal microscopy techniques. Confocal microscopy evaluations showed that there was no noteworthy amount of BMSCs in the lung tissue of group III while significant amount of BMSCs was detected in group II. Wall thicknesses of terminal bronchiole and periterminal bronchiolar collagen deposition were significantly lower in group II compared to the group IV (p<0.05). Furthermore, according to the immunohistochemical staining results, CD3, CD4, CD8, CD20, CD68 and neutrophil elastase positive immune cells of group II were stained more positive than group IV cells (p<0.05). IFN-γ IL-2 and TNF-α levels in bronchoalveolar lavage fluid (BALF) were significantly lower in group II compared to group IV (p<0.05). The findings of this study indicate that intraperitoneally administered BMSCs have potent effects on histopatological changes of the lung tissue and cytokine levels in the murine model of BO.


Hemagglutinin stalk-based universal vaccine constructs protect against group 2 influenza A viruses.
Current influenza virus vaccines contain H1N1 (phylogenetic group 1 hemagglutinin), H3N2 (phylogenetic group 2 hemagglutinin), and influenza B virus components. These vaccines induce good protection against closely matched strains by predominantly eliciting antibodies against the membrane distal globular head domain of their respective viral hemagglutinins. This domain, however, undergoes rapid antigenic drift, allowing the virus to escape neutralizing antibody responses. The membrane proximal stalk domain of the hemagglutinin is much more conserved compared to the head domain. In recent years, a growing collection of antibodies that neutralize a broad range of influenza virus strains and subtypes by binding to this domain has been isolated. Here, we demonstrate that a vaccination strategy based on the stalk domain of the H3 hemagglutinin (group 2) induces in mice broadly neutralizing anti-stalk antibodies that are highly cross-reactive to heterologous H3, H10, H14, H15, and H7 (derived from the novel Chinese H7N9 virus) hemagglutinins. Furthermore, we demonstrate that these antibodies confer broad protection against influenza viruses expressing various group 2 hemagglutinins, including an H7 subtype. Through passive transfer experiments, we show that the protection is mediated mainly by neutralizing antibodies against the stalk domain. Our data suggest that, in mice, a vaccine strategy based on the hemagglutinin stalk domain can protect against viruses expressing divergent group 2 hemagglutinins.


Endocrine regulation of fetal growth.
Hormones have an important role in the control of fetal growth. They act on both tissue accretion and differentiation and enable a precise and orderly pattern of growth to occur during late gestation. In part, their actions on growth may be mediated by other growth factors such as the insulin-like growth factors (IGFs). Insulin stimulates fetal growth by increasing the mitotic drive and nutrient availability for tissue accretion. It has little effect on tissue differentiation. In contrast, the main effects of cortisol in utero are on tissue differentiation and maturation. Cortisol appears to act directly on the cells to alter gene transcription or post-translational processing of the gene products. Cortisol may also initiate the transition from the fetal to the adult modes of growth regulation by inducing the switch from IGF-II to IGF-I gene expression in the fetal liver. Thyroxine affects both tissue accretion and differentiation in the fetus by a combination of metabolic and non-metabolic mechanisms. Pituitary growth hormone, on the other hand, appears to have little part in the control of fetal growth, unlike its role postnatally. Fetal hormones, therefore, promote growth and development in utero by altering both the metabolism and gene expression of the fetal tissues. These hormonal actions ensure that fetal growth rate is commensurate with the nutrient supply and that prepartum maturation occurs in preparation for extrauterine life.


Stakeholder and public participation in river basin management-an introduction.
Participation of the public and stakeholders in river basin management is increasingly promoted because it is expected to improve resource management and enable participants to engage freely and equally in management (support democratic processes). Three overlapping and interacting mechanisms by which participation is expected to enhance river basin management are outlined: (1) providing space for deliberation and consensus building for better quality decisions, (2) mobilizing and developing human and social capital for better quality decisions and their implementation, and (3) raising the legitimacy of decisions to facilitate their implementation. There are several complexities associated with each of the mechanisms that add challenges to realizing the expectations of participation. They include the need to carefully manage consensus building and conflict to maximize the quality of the decision without jeopardizing the potential for implementation; being aware of and implementing strategies to manage asymmetrical power relationships between participants; ensuring that participants perceive benefits from participation that exceed costs; and defining criteria for a legitimate process, and a legitimate decision, that satisfy all participants. Strategies identified to address these challenges focus on managing the characteristics of the participation process. Ongoing evaluation during a participation program or project is essential to reflect and refine how participation is being done, to address the challenges and endeavor to achieve high-quality decisions that can be implemented efficiently. WIREs Water 2015, 2:393-405. doi: 10.1002/wat2.1086 For further resources related to this article, please visit the WIREs website.


Efficacy of a probiotic and chlorhexidine mouth rinses: a short-term clinical study.
Probiotic technology represents a breakthrough approach to maintaining oral health by utilizing natural beneficial bacteria commonly found in healthy mouths to provide a natural defense against those bacteria thought to be harmful to teeth and gums. However, data are still sparse on the probiotic action in the oral cavity. The review article on probiotics in children published by Twetman and Stecksen- Blicks in 2008 showed only one study of dental interest on probiotics in children. The present study evaluated clinically the efficacy of a probiotic and chlorhexidine mouth rinses on plaque and gingival accumulation in children. The trial design is a double-blind parallel group, 14 days comparative study between a probiotic mouth rinse and a chlorhexidine mouth rinse, which included 45 healthy children in the age group of 6-8 years. The Probiotic and Chlorhexidine groups had less plaque accumulations compared with the Control group at the end of 14 years (P < 0.001 and P < 0.001, respectively). But, unlike the plaque score, there was a significant difference in the Gingival Index between the Probiotic and the Chlorhexidine groups (P = 0.009), Probiotic group being better than the Chlorhexidine group (mean = 0.2300 and 0.6805, respectively). The Probiotic mouth rinse was found effective in reducing plaque accumulation and gingival inflammation. Therefore, probiotic mouth rinse obviously has a potential therapeutic value and further long-term study is recommended to determine its efficacy.


A chemically-defined medium for the growth of a ureolytic strain of Streptococcus faecium.
A chemically-defined medium was developed which supported growth of Streptococcus faecium and permitted synthesis of urease. This streptococcus cannot utilize ammonia and needs a complex medium, but its requirements are probably provided in the rumen. The specific activity of urease was inversely related to growth and in no medium was there high growth and high urease activity. Anaerobic culture and the presence of urea in the medium were essential for urease activity, but not for growth.


Hypermethylation of MIR21 in CD4+ T cells from patients with relapsing-remitting multiple sclerosis associates with lower miRNA-21 levels and concomitant up-regulation of its target genes.
Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system caused by genetic and environmental factors. DNA methylation, an epigenetic mechanism that controls genome activity, may provide a link between genetic and environmental risk factors. We sought to identify DNA methylation changes in CD4+ T cells in patients with relapsing-remitting (RR-MS) and secondary-progressive (SP-MS) disease and healthy controls (HC). We performed DNA methylation analysis in CD4+ T cells from RR-MS, SP-MS, and HC and associated identified changes with the nearby risk allele, smoking, age, and gene expression. We observed significant methylation differences in the VMP1/MIR21 locus, with RR-MS displaying higher methylation compared to SP-MS and HC. VMP1/MIR21 methylation did not correlate with a known MS risk variant in VMP1 or smoking but displayed a significant negative correlation with age and the levels of mature miR-21 in CD4+ T cells. Accordingly, RR-MS displayed lower levels of miR-21 compared to SP-MS, which might reflect differences in age between the groups, and healthy individuals and a significant enrichment of up-regulated miR-21 target genes. Disease-related changes in epigenetic marking of MIR21 in RR-MS lead to differences in miR-21 expression with a consequence on miR-21 target genes.


Engineered modular recombinant transporters: application of new platform for targeted radiotherapeutic agents to alpha-particle emitting 211 At.
To generate and evaluate a modular recombinant transporter (MRT) for targeting 211 At to cancer cells overexpressing the epidermal growth factor receptor (EGFR). The MRT was produced with four functional modules: (1) human epidermal growth factor as the internalizable ligand, (2) the optimized nuclear localization sequence of simian vacuolating virus 40 (SV40) large T-antigen, (3) a translocation domain of diphtheria toxin as an endosomolytic module, and (4) the Escherichia coli hemoglobin-like protein (HMP) as a carrier module. MRT was labeled using N-succinimidyl 3-[211 At]astato-5-guanidinomethylbenzoate (SAGMB), its 125 I analogue SGMIB, or with 131 I using Iodogen. Binding, internalization, and clonogenic assays were performed with EGFR-expressing A431, D247 MG, and U87MG.wtEGFR human cancer cell lines. The affinity of SGMIB-MRT binding to A431 cells, determined by Scatchard analysis, was 22 nM, comparable to that measured before labeling. The binding of SGMIB-MRT and its internalization by A431 cancer cells was 96% and 99% EGFR specific, respectively. Paired label assays demonstrated that compared with Iodogen-labeled MRT, SGMIB-MRT and SAGMB-MRT exhibited more than threefold greater peak levels and durations of intracellular retention of activity. SAGMB-MRT was 10-20 times more cytotoxic than [211 At]astatide for all three cell lines. The results of this study have demonstrated the initial proof of principle for the MRT approach for designing targeted alpha-particle emitting radiotherapeutic agents. The high cytotoxicity of SAGMB-MRT for cancer cells overexpressing EGFR suggests that this 211 At-labeled conjugate has promise for the treatment of malignancies, such as glioma, which overexpress this receptor.


Networks in cognitive systems and biomedicine: cerebral processes, models and mathematical tools design.
Convergence of clinical, empirical, methodological and theoretical approaches aimed at understanding the relation between brain function and cognition, is by now standard in most if not all academic programs in the area of Cognitive Science. This confederation of disciplines is one of the liveliest domains of inquiry and discussion into some of the most fundamental--and historically resilient--questions human beings have posed themselves. The contributions gathered in this special issue of Biological Research, directly inspired by the ongoing work at the Instituto de Sistemas Complejos de Valparaiso and the December 2006 CONICYT-INSERM-SFI workshop "Networks in Cognitive Systems/Trends and Challenge in Biomedicine: From Cerebral Process to Mathematical Tools Design", Chile, represent an explicit invitation to the reader to dive deeper into this fascinating terrain.


Dynamic systems theory as a paradigm for the study of change in psychotherapy: an application to cognitive therapy for depression.
Dynamic systems theory provides a conceptual framework for the study of change in psychotherapy that is consistent with that used in other sciences. A dynamic systems model of change was proposed and evaluated in the context of cognitive therapy for depression. Consistent with this model, less client protection and more destabilization of depressive patterns predicted more improvement at the end of treatment. Less protection was associated with more therapist support/stabilization. More destabilization was associated with more affective intensity in the session and with more of a therapist focus on the historical antecedents of current problems, exposure to multiple sources of corrective information, and repeated practice of new skills. Although preliminary, this pattern of findings is consistent with the model proposed and with principles of dynamic systems from other sciences.


Satisfaction with civilian family medicine residency training: Perspectives from serving general duty medical officers in the Canadian Armed Forces.
To evaluate satisfaction with civilian residency training programs among serving general duty medical officers within the Canadian Armed Forces. A 23-item, cross-sectional survey face-validated by the office of the Surgeon General of the Canadian Armed Forces. Canada. General duty medical officers serving in the Canadian Armed Forces as of February 2014 identified through the Directorate of Health Services Personnel of the Canadian Forces Health Services Group Headquarters. Satisfaction with and time spent in 7 domains of training: trauma, critical care, emergency medicine, psychiatry, occupational health, sports medicine, and base clinic training. Overall preparedness for leading a health care team, caring for a military population, working in isolated and challenging environments, and being deployed were evaluated on a 5-point Likert scale. Among the survey respondents (n = 135, response rate 54%), 77% agreed or strongly agreed that their family medicine residency training was relevant to their role as a general duty medical officer. Most respondents were either satisfied or very satisfied with their emergency medicine training (77%) and psychiatry training (63%), while fewer were satisfied or very satisfied with their sports medicine (47%), base clinic (41%), and critical care (43%) training. Even fewer respondents were satisfied or very satisfied with their trauma (26%) and occupational health (12%) training. Regarding overall preparedness, 57% believed that they were adequately prepared to care for a military patient population, and 52% of respondents believed they were prepared for their first posting. Fewer respondents (38%) believed they were prepared to work in isolated, austere, or challenging environments, and even fewer (32%) believed that residency training prepared them to lead a health care team. General duty medical officers were satisfied with many aspects of their family medicine residency training; however, military-specific areas for improvement were identified. Many of these areas might be addressed within the context of a 2-year residency program without risking the generalist nature of family medicine training. These findings provide valuable data for residency programs that accept military trainees across the country.


Intrafasciomembranal Fluid Pressure: A Novel Approach to the Etiology of Myalgias.
Fascia is a continuous membrane (fasciomembrane) that enables differentiation of fluid pressure on either side. Fascia membrane also enables an internal increased fluid pressure at all muscle levels (fibers, fiber bundles, skeletal muscles, compartments), and the author introduces a new unifying term for these pressures, regardless of the anatomical level - the intrafasciomembranal fluid pressure (IFMFP). Swelling, pain, and loss of tissue function are identified as common cardinal symptoms in trigger point (TrP), chronic exertional compartment syndrome (CECS), overtraining syndrome (OTS), and delayed onset muscle soreness (DOMS). Existing literature and an overall assessment indicate that intramuscular conditions related to fluid flow and pressure play a central role in different conditions, providing a common biomechanical explanation of the etiology and influence, supporting the article's theory that an increased IFMFP plays a key role in these conditions.


The STAT5 inhibitor pimozide decreases survival of chronic myelogenous leukemia cells resistant to kinase inhibitors.
The transcription factor STAT5 is an essential mediator of the pathogenesis of chronic myelogenous leukemia (CML). In CML, the BCR/ABL fusion kinase causes the constitutive activation of STAT5, thereby driving the expression of genes promoting survival. BCR/ABL kinase inhibitors have become the mainstay of therapy for CML, although CML cells can develop resistance through mutations in BCR/ABL. To overcome this problem, we used a cell-based screen to identify drugs that inhibit STAT-dependent gene expression. Using this approach, we identified the psychotropic drug pimozide as a STAT5 inhibitor. Pimozide decreases STAT5 tyrosine phosphorylation, although it does not inhibit BCR/ABL or other tyrosine kinases. Furthermore, pimozide decreases the expression of STAT5 target genes and induces cell cycle arrest and apoptosis in CML cell lines. Pimozide also selectively inhibits colony formation of CD34(+) bone marrow cells from CML patients. Importantly, pimozide induces similar effects in the presence of the T315I BCR/ABL mutation that renders the kinase resistant to presently available inhibitors. Simultaneously inhibiting STAT5 with pimozide and the kinase inhibitors imatinib or nilotinib shows enhanced effects in inhibiting STAT5 phosphorylation and in inducing apoptosis. Thus, targeting STAT5 may be an effective strategy for the treatment of CML and other myeloproliferative diseases.


High-speed slot-scanning radiography using small-angle tomosynthesis: Investigation of spatial resolution.
This paper studies spatial resolution that is achievable with a fast slot-scanning tomosynthesis approach for orthopedic examinations. Hereby, we use parallel scanning motion implemented in a twin robotic x-ray system. We have measured and analyzed the modulation transfer function (MTF) for various combinations of scanning speed, x-ray tube voltage, pulse length, the nominal focal spot size as well as source-to-object distances. Moreover, we present a theoretical model which describes the system in terms of the MTF. The system was equipped with newly developed linear trajectory prototypes for slot scanning. The acquired images form the basis for a small-angle tomosynthesis reconstruction. In total, three different scanning speeds (27, 14, 8 cm/s), pulse lengths (1, 2, 4 ms), tube voltages (80, 100, 120 kV), two nominal focal spot sizes (0.6, 1.0), and three source-to-object distances (950, 1050, 1150 mm) were investigated. To determine the resolution capabilities, we measured the MTF for the given parameter space. The results were then used to design a filter that yields a desired resolution in the reconstructed image. In addition, we also measured the noise power spectrum (NPS) to show the influence of the aforementioned filters on the noise distribution. We have shown that the presented model is in good agreement with the performed measurements. Scanning speed and pulse width have an impact on the MTF in the scanning direction. Up to a travel distance of 0.3 mm during an x-ray pulse, an isotropic resolution can be achieved. Longer pulse width or higher scanning speed cause anisotropic resolution. Moreover, it is shown that none of the investigated parameters have an influence on the MTF perpendicular to the scanning direction (slot direction). The 10% MTF value ranges between 9 and 18 lp/cm in the scanning direction and about 18 lp/cm in slot direction. Tube voltage, nominal focal spot size as well as the source-to-object distance showed no major impact on the system MTF. In terms of the anisotropic resolution capabilities, we have shown that limiting the resolution in the slot direction to obtain isotropic resolution is possible yet at the cost of an inhomogeneous noise pattern. However, maintaining the resolution in slot direction will provide a better edge response and a more homogeneous noise texture at the cost of an inhomogeneous image resolution. We have demonstrated the feasibility of the slot-scanning technique using a twin-robotic x-ray system. Even the fastest scanning mode (27 cm/s) yields image resolution on a level that is sufficient for typical orthopedic examinations in terms of musculoskeletal (MSK) measurements. Moreover, it could be shown that the application of specifically designed target MTFs on two-dimensional x-ray images is feasible.


[Kinetic characteristics of the ATP-pyrophosphate isotope exchange catalyzed by RNA ligase from T4 bacteriophage].
The dependence of initial rate v0 of ATP--PPi exchange reaction catalyzed by RNA-ligase of bacteriophage T4 on the concentration of ATP(s), pyrophosphate (z) and Mgcl2 has been determined. The dependence of v0 on s and z described by the equation v0 = k-1k2E0/(k-1 + K2) (1 + K1/s + k2/z) has been obtained for the reaction of E + S in equilibrium ES in equilibrium E1 + Z, where E--enzyme, E1--adenylylenzyme, S--ATP, Z--pyrophosphate, K1 and K2--constants of equilibrium, k-1, k2--velocity constants of transition of ES to E + S and E1 + Z, E0--complete concentration of enzyme. The low inhibition of the ATP--PPi exchange by the acceptor A(pA)2 and donors pAp, p(Ap)3, pCp has been shown. The dependence of v0 on the concentration of MgCl2 is consent with the incorporation of only dimagnesium salts of substrates in the isotope-exchange reaction.


Highly sensitive serum thyroglobulin and circulating thyroglobulin mRNA evaluations in the management of patients with differentiated thyroid cancer in apparent remission.
We studied the potential role of innovative diagnostic tools for the management of patients with differentiated thyroid cancer (DTC). Several methods for the detection of the tumor marker thyroglobulin (Tg) have been employed in 36 patients in apparent remission at the moment of the study. All patients had negative anti-Tg antibodies and were evaluated during L-T(4) suppressive therapy before and after stimulation with recombinant human TSH (rhTSH). Serum Tg was measured by means of conventional [nonhighly sensitive (nhs)] or highly sensitive (hs) immunoassays with positive cut-off values set at 1.0 and 0.18 microg/liter, respectively. The RT-PCR conditions for the qualitative determination of Tg mRNA from peripheral blood were optimized to prevent interference by illegitimate transcription. The patients have been classified on the basis of a hs-basal Tg testing by taking into account the results of their baseline samples in hs immunoassay and RT-PCR method; hs-basal Tg testing was considered positive when the marker was detectable in at least one of the two tests. The predictive value of hs-basal Tg testing was estimated on the basis of a global clinical evaluation, including serum Tg response after rhTSH stimulation and reports of contemporary (131)I scan and neck ultrasound. The clinical evaluation was considered positive when at least one of these criteria yielded positive results. Although nhs-Tg measurement was poorly predictive of the clinical status, basal hs-Tg evaluation was found to be concordant with the clinical evaluation in 71% of cases. Results of basal Tg mRNA detection did not vary after rhTSH stimulation and were concordant with the clinical evaluation in 66% of cases. Tg mRNA evaluation alone showed 10 apparently false-positive results, and serum basal hs-Tg was falsely negative in 11 additional cases, suggesting that a suitable predictability could be obtained by the association of these 2 parameters. Indeed, the combination of hs-Tg assay and mRNA detection in the hs-basal Tg testing allowed the identification of 22 patients with a positive persistent/recurrent disease or normal thyroid residue, as well as identification of all 6 patients with a negative clinical evaluation. In conclusion, the combined evaluation of circulating Tg mRNA and serum Tg by means of hs noncompetitive immunoassay (hs-basal Tg testing) can give useful information on the clinical status of patients with DTC who are apparently disease-free, even on L-T(4) TSH-suppressive therapy. Therefore, these combined evaluations retain a potential role in the clinical monitoring of DTC patients. In particular, a negative hs-basal Tg testing would indicate disease remission and the opportunity to lengthen the intervals between rhTSH stimulations and/or to shift patients to a less profound TSH suppression with L-T(4).


Antidepressants and chronic pain.
Successful treatment of pain syndromes is one of the most common and most difficult problems facing family physicians. Frequently analgesics provide inadequate treatment, and clinicians are forced to consider alternatives. This article reviews the neurophysiologic similarities between depression and the chronic pain syndromes and describes several well-designed double-blinded studies that give evidence for the efficacy of antidepressants in chronic pain syndromes. These studies conclude that antidepressants should be considered in chronic pain syndromes that do not respond to analgesics. For chronic pain, antidepressants should be started at a low dosage and increased in a stepwise manner until an improvement in the pain occurs or intolerable side effects intervene. Side effects are a bothersome aspect of antidepressant therapy but are more tolerable at the doses generally needed for pain relief than at antidepressive doses. At least three weeks of antidepressant therapy is generally needed to gain significant relief of symptoms.


Accelerating molecular dynamic simulation on graphics processing units.
We describe a complete implementation of all-atom protein molecular dynamics running entirely on a graphics processing unit (GPU), including all standard force field terms, integration, constraints, and implicit solvent. We discuss the design of our algorithms and important optimizations needed to fully take advantage of a GPU. We evaluate its performance, and show that it can be more than 700 times faster than a conventional implementation running on a single CPU core.


Elucidating the structure of carbon nanoparticles by ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry.
A fast and accurate ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) method was developed for the separation and structural elucidation of fluorescent carbon nanoparticles (CNP). The CNP was synthesised from microwave-assisted pyrolysis of citric acid (CA) and 1,2-ethylenediamine (EDA). By using UPLC separation, the CNP product was well separated into ten fractions within 4.0 min. Based on high-accuracy MS and MS/MS analyses, the CNP species were revealed to display six kinds of chemical formulas, including (C10H20N4O5)n, (C8H12N2O5)n, (C16H22N4O9)n, (C6H8O7)n, (C14H18N2O11)n, and (C14H16N2O10)n. In particular, our study revealed for the first time that the CNP species exist as supramolecular clusters with their individual monomers units linked together through non-covalent bonding forces. These findings clearly indicated the usefulness of UPLC-ESI-Q-TOF-MS/MS in identifying the chemical composition of CNP product. It is anticipated that our proposed methodology can be applied to study the structure-property relationships of CNP, facilitating in the production of CNP with desirable spectral features.


Actions of human interleukin-4/B-cell stimulatory factor-1 on proliferation and differentiation of enriched hematopoietic progenitor cells in culture.
We studied the effects of recombinant human interleukin-4 (IL-4) on colony formation by enriched hematopoietic progenitors. IL-4 alone did not support colony formation at all. When IL-4 was combined with granulocyte colony-stimulating factor (G-CSF), the number of pure neutrophil colonies increased three times over that supported by G-CSF alone. IL-4 added 5 days after the addition of G-CSF failed to exert this synergistic effect, indicating that IL-4 acts on the early stage of proliferation. The mapping experiments (sequential observation of colony formation) have clearly shown that IL-4 did not initiate progenitor cell proliferation. Based on these data, IL-4 may possess a direct action on progenitor cells; however, it can only act as a costimulant with G-CSF. In contrast, IL-4 had possible inhibitory effects on macrophage colony formation supported by interleukin-3 (IL-3) and macrophage colony-stimulating factor (M-CSF). In other words, IL-4 may induce progenitor cells to become sensitive to G-CSF and thereby induce neutrophil differentiation. Delayed addition experiments demonstrated that human IL-4, unlike murine IL-4, could support neither proliferation nor survival of erythroid burst or mixed colony forming cells. Neutrophil colony forming cells only survived and recovered after addition of G-CSF and erythropoietin on day 5 of incubation. On the other hand, IL-3 supported neutrophil, erythroid burst, and mixed colony forming cells as reported previously (Sonoda et al, Proc Natl Acad Sci USA, 85:4360, 1988). These results led us to propose that IL-4 possibly acts with more mature progenitor cells than those of IL-3 or granulocyte-macrophage (GM)-CSF.


Adherence to Objective Therapeutic Monitoring and Outcomes in Patients with Inflammatory Bowel Disease with Adalimumab Treatment. A Real-world Prospective Study.
Objective monitoring and effective early treatment using a treat-to-target approach are key to improving therapeutic outcomes in IBD patients. This study aimed to assess adherence to objective monitoring (clinical, biomarkers, and endoscopy) and its impact on clinical outcomes. A prospective, multicenter study included consecutive IBD patients starting on adalimumab therapy between January 2019 and December 2020. Disease activity, assessed by the Harvey-Bradshaw index (HBI), partial Mayo, C-reactive protein (CRP), fecal calprotectin (FCAL), and endoscopy were evaluated at adalimumab initiation and 3, 6, 9 and 12 months. Therapeutic drug monitoring, changes in treatment, drug sustainability, and clinical outcomes were assessed. 104 IBD patients were enrolled (78.8% CD, median age 34.3 years, disease duration 9 years). During the 12 months follow-up, high adherence to clinical activity assessment was observed in both CD (81.3%- 87.7%) and UC patients (76.5-90.9%). CRP measurement decreased over time in both CD (37.3%-54.9%) and UC (29.4%-50.0%). The adherence to serial FCAL monitoring was low in CD (22.7-31.3%) and UC patients (17.6-56.0%). UC patients had higher adherence to early endoscopic assessment (<6 months) compared to CD patients (40.9% vs. 21.5%). Adherence to early combined clinical and biomarkers resulted in earlier dose optimization in CD and UC (log-rank<0.001), but drug sustainability was not different. The patients with early combined adherence had a significantly higher clinical remission rate at 1 year compared to non-adherence (70.2% vs. 29.8%, p=0.007) but no significant difference in UC patients. The adherence to early objective monitoring with combined clinical and biomarkers assessment in IBD patients starting adalimumab therapy led to dose optimization and improved 1-year clinical remission in CD but did not change drug sustainability and clinical remission in UC.


[Spontaneous hematoma of the cerebellum].
Forty spontaneous cerebellar hematomas have been observed in 39 patients whose age ranged from 13 to 82 years, 26 being older than 60. Arterial hypertension is the most frequent aetiological factor. Amongst the other causes, vascular malformations, which may also be present in older people, anticoagulant therapy, metastatic tumor, sepsis were found; one case remained unsolved. Diagnosis is no longer a challenge since the introduction of C.T. scan, whilst M.R.I. may give most valuable informations concerning aetiology, especially when a vascular malformation is present. In this study, the influence of the hematoma size and of hydrocephalus on consciousness was demonstrated; development of hydrocephalus is favoured by the size and by a rather median location of the hematoma. In the future, M.R.I. should give better precisions concerning an eventual extension of the hemorrhage to the brainstem. The purpose of surgical treatment is twofold: alleviating brainstem compression and correcting hydrocephalus; yet, divergent opinions have been put forward in retrospective studies, concerning evacuation of the hematoma and drainage of hydrocephalus. In this series, direct attack doesn't seem to be indicated in hematomas less than 3.5 cm diameter, unless a causal lesion has to be removed. The ideal moment for hematoma evacuation is, finally, dictated by the clinical status and its evolution; however, the authors prefer, if possible, to delay it for at least 48 hours. Amongst the 39 patients, 14 deceased; of the 25 survivors, 19 were autonomous, 7 being nearly asymptomatic, 6 patients remained handicapped. Consciousness is a main prognostic factor, whilst coma in the first hours has a clearly unfavourable significance.


Energy expenditure in individuals with spinal cord injury quantified by doubly labeled water and a multi-sensor armband.
The objective of this study was to determine whether doubly labeled water (DLW) and a multi-sensor armband (SWA) could detect the variation in energy expenditure incurred by a period of increased exercise (EXE) versus a period of high sedentary activity (SED), in individuals with spinal cord injury (SCI). Eight participants with SCI were submitted to 2 testing periods of energy expenditure assessment: 1) a 14-day phase during which sedentary living conditions were imposed and 2) a 14-day phase during which an exercise training intervention was employed. For each phase, total daily energy expenditure (TDEE) and physical activity energy expenditure (PAEE) were measured by DLW and SWA. Mean TDEE assessed by DLW, was significantly higher during EXE versus SED (11,605 ± 2151 kJ·day⁻¹ and 10,069 ± 2310 kJ·day⁻¹). PAEE predicted by DLW was also significantly higher during EXE versus SED (5422 ± 2240 kJ·day⁻¹ and 3855 ± 2496 kJ·day⁻¹). SWA-predicted PAEE significantly underestimated PAEE measured by the DLW during SED and EXE. DLW is sensitive to detect variation in within-individual energy expenditure during voluntary increase in physical activity in individuals with SCI. SWA failed to detect statistically significant variations in energy expenditure between periods of high versus low activity in SCI.


Phaeohyphomycosis caused by Diaporthe phaseolorum in an immunocompetent patient in Thailand: a case report.
Phaeohyphomycosis is caused by a large, heterogeneous group of darkly pigmented fungi. It is an infrequent infection in humans. However, the prevalence has been increasing in recent years especially in immunocompromised patients. Diaporthe phaseolorum is a common black fungal pathogen of plants, which rarely causes human infection. We report the first case of cutaneous infection caused by Diaporthe phaseolorum in an immunocompetent host and the first in Asia. Although, the review of the literature revealed two previous cases of cutaneous infection caused by this organism, both of them were in immunocompromised hosts. A slow-growing asymptomatic nodule was the major clinical feature. Histopathological examination showed granulomatous inflammation and pigmented septate hyphae and yeast-like cells. The fungal isolation was identified by morphological characteristics and DNA sequencing. The lesion was resolved after complete surgical excision and oral fluconazole for two months. This report highlights the potential role of Diaporthe phaseolorum as an emerging cause of infection in immunocompetent patients.


Impact of diabetes and peripheral arterial occlusive disease on the functional microcirculation at the plantar foot.
Plastic and reconstructive surgeons are commonly faced with chronic ulcerations and consecutive wound infections of the feet as complications in patients with diabetes and/or peripheral arterial occlusive disease (PAOD). Microcirculatory changes seem to play an important role. However, the evaluation of functional changes in the soft tissue microcirculation at the plantar foot using combined Laser-Doppler and Photospectrometry System has not yet been performed in patients with DM or PAOD. A prospective, controlled cohort study was designed consisting of a total of 107 subjects allocated to 1 of 3 groups-group A: healthy subjects (57% males, 63.3 y); group B: patients with diabetes mellitus (DM) (53% males, 59.4 y); and group C: patients with PAOD (81% males, 66.1 y). Microcirculatory data were assessed using a combined Laser-Doppler and Photospectrometry System. Global cutaneous oxygen saturation microcirculation at the plantar foot of healthy individuals was 8.4% higher than in patients with DM and 8.1% higher than in patients with PAOD (both P = 0.033). Patients with diabetes did not show significant differences in global cutaneous blood flow when compared with either healthy subjects or patients suffering from PAOD. Functional microcirculation at the plantar foot differs between healthy subjects and patients suffering from diabetes or PAOD of the same age. Patients with either diabetes or PAOD demonstrate deteriorated cutaneous oxygen saturation with equivalent blood perfusion at the plantar foot. More clinical studies have to be conducted to evaluate therapeutical methods that might ameliorate cutaneous oxygen saturation within diabetic foot disease and PAOD.


Sexual risk behavior and sex under the influence: an event analysis of men in substance abuse treatment who have sex with women.
The objective of this study was to determine if there is evidence for a causative link between sex under the influence of drugs or alcohol and risky sex for men in substance abuse treatment. Men in treatment participating in a multisite HIV prevention protocol who reported on baseline, 3, or 6 months computerized assessments the details of their most recent sexual events, and who reported having sexual events under the influence and not under the influence, and who reported most recent events that did and did not include condom use served as participants (n = 37). Safe sex was not significantly more likely to happen when participants were under the influence of drugs or alcohol during their most recent sexual event (48.3%) than when they were not under the influence (49%, p = .82). In this high-risk in treatment sample, a causative link between sex under the influence of drugs or alcohol and sexual risk behavior was not supported.


Population based absolute and relative survival to 1 year of people with diabetes following a myocardial infarction: a cohort study using hospital admissions data.
People with diabetes who experience an acute myocardial infarction (AMI) have a higher risk of death and recurrence of AMI. This study was commissioned by the Department for Transport to develop survival tables for people with diabetes following an AMI in order to inform vehicle licensing. A cohort study using data obtained from national hospital admission datasets for England and Wales was carried out selecting all patients attending hospital with an MI for 2003-2006 (inclusion criteria: aged 30+ years, hospital admission for MI (defined using ICD 10 code I21-I22). STATA was used to create survival tables and factors associated with survival were examined using Cox regression. Of 157,142 people with an MI in England and Wales between 2003-2006, the relative risk of death or recurrence of MI for those with diabetes (n = 30,407) in the first 90 days was 1.3 (95%CI: 1.26-1.33) crude rates and 1.16 (95%CI: 1.1-1.2) when controlling for age, gender, heart failure and surgery for MI) compared with those without diabetes (n = 129,960). At 91-365 days post AMI the risk was 1.7 (95% CI 1.6-1.8) crude and 1.50 (95%CI: 1.4-1.6) adjusted. The relative risk of death or re-infarction was higher at younger ages for those with diabetes and directly after the AMI (Relative risk; RR: 62.1 for those with diabetes and 28.2 for those without diabetes aged 40-49 [compared with population risk]). This is the first study to provide population based tables of age stratified risk of re-infarction or death for people with diabetes compared with those without diabetes. These tables can be used for giving advice to patients, developing a baseline to compare intervention studies or developing license or health insurance guidelines.


Effective surface conductivity of optical hyperbolic metasurfaces: from far-field characterization to surface wave analysis.
Metasurfaces offer great potential to control near- and far-fields through engineering optical properties of elementary cells or meta-atoms. Such perspective opens a route to efficient manipulation of the optical signals both at nanoscale and in photonics applications. In this paper we show that a local surface conductivity tensor well describes optical properties of a resonant plasmonic hyperbolic metasurface both in the far-field and in the near-field regimes, where spatial dispersion usually plays a crucial role. We retrieve the effective surface conductivity tensor from the comparative analysis of experimental and numerical reflectance spectra of a metasurface composed of elliptical gold nanoparticles. Afterwards, the restored conductivities are validated by semi-analytic parameters obtained with the nonlocal discrete dipole model with and without interaction contribution between meta-atoms. The effective parameters are further used for the dispersion analysis of surface plasmons localized at the metasurface. The obtained effective conductivity describes correctly the dispersion law of both quasi-TE and quasi-TM plasmons in a wide range of optical frequencies as well as the peculiarities of their propagation regimes, in particular, topological transition from the elliptical to hyperbolic regime with eligible accuracy. The analysis in question offers a simple practical way to describe properties of metasurfaces including ones in the near-field zone with effective conductivity tensor extracting from the convenient far-field characterization.


Approach to premenstrual dysphoria for the mental health practitioner.
As many as 7% of women experience significant social or occupational dysfunction as a result of severe premenstrual mood disturbance. Biological, psychological, and sociocultural factors are implicated in the cause of premenstrual dysphoric disorder, but the interaction between these factors remains to be elucidated. Mental health practitioners can aid women by providing diagnostic clarity and by initiating an integrated step-wise management approach.


Effects of previous activity on the energetics of activation in frog skeletal muscle.
Effects of previous activity on the ability of frog skeletal muscle at 0 degrees C to liberate energy associated with contractile activation, i.e., activation heat (AH), have been examined. Earlier work suggests that activation heat amplitude (as measured from muscles stretched to lengths where active force development is nearly abolished) is related to the amount of Ca2+ released upon stimulation. After a twitch, greater than 2 s is required before a second stimulus (AHt) can liberate the same activation heat as a first stimulus (AH infinity), i.e., (AHt)/(AH infinity) = 1 -0.83 e-1.40t, where t is time in seconds. Caffeine introduces a time delay in the recovery of the ability to generate activation heat after a twitch. After a tetanus, the activation heat is depressed to a greater extent at any time than after a twitch. The activation heat elicited by a stimulus 1 s after a tetanus is depressed progressively with respect to tetanus duration up to 3 s. For tetani of 3, 40, and 80 s duration the postetanus activation heat is comparably depressed. The time-course of the recovery of the ability of the muscle to produce activation heat after a tetanus can be described as (AHt)/(AH infinity) = 1 -0.80 e-0.95t -0.20 e-0.02t. Greater than 90 s is required before the posttetanus activation heat is equal to the pretetanus value. The faster phase of recovery is similar to recovery after the twitch and the slower phase may be associated with the return of calcium to the terminal cisternae from uptake sites in the longitudinal sarcoplasmic reticulum.


Unchanged muscle fiber conduction velocity relates to mild acidosis during exhaustive bicycling.
Muscle fiber conduction velocity (MFCV) has often been shown to decrease during standardized fatiguing isometric contractions. However, several studies have indicated that the MFCV may remain constant during fatiguing dynamic exercise. It was investigated if these observations can be related to the absence of a large decrease in pH and if MFCV can be considered as a good indicator of acidosis, also during dynamic bicycle exercise. High-density surface electromyography (HDsEMG) was combined with read-outs of muscle energetics recorded by in vivo (31)P magnetic resonance spectroscopy (MRS). Measurements were performed during serial exhausting bouts of bicycle exercise at three different workloads. The HDsEMG recordings revealed a small and incoherent variation of MFCV during all high-intensity exercise bouts. (31)P MRS spectra revealed a moderate decrease in pH at the end of exercise (~0.3 units down to 6.8) and a rapid ancillary drop to pH 6.5 during recovery 30 s post-exercise. This additional degree of acidification caused a significant decrease in MFCV during cycling immediately after the rest period. From the data a significant correlation between MFCV and [H(+)] ([H(+)] = 10(-pH)) was calculated (p < 0.001, Pearson's R = -0.87). Our results confirmed the previous observations of MFCV remaining constant during fatiguing dynamic exercise. A constant MFCV is in line with a low degree of acidification, considering the presence of a correlation between pH and MFCV after further increasing acidification.


Cloning. Pigs is pigs.
Since the first report of a cloned animal (Dolly the sheep) 3 years ago, cloning mammals has become something of a cottage industry. As Prather discusses in his Perspective, pigs can now be added to the august list of cloned animals, which includes cows, goats, and mice. This is a particularly spectacular achievement because pig cloning has turned out to be notoriously difficult. The pig is also a valuable domestic animal to have cloned because, being physiologically close to humans, its organs can be used in xenotransplantation.


Cross-reactivity of the anti-La monoclonal antibody SW5 with early endosome antigen 2.
Coimmunoprecipitation studies with SW5, a frequently used and specific mouse monoclonal antibody (mAb) directed against the human La autoantigen, led to the identification of a functionally unrelated 80 000 MW protein, designated early endosome antigen 2 (EEA2). EEA2 appeared to be directly targeted by mAb SW5. Because an RNA-binding domain, a structural element of La containing the SW5-epitope, was not discernable in the primary structure of EEA2, the SW5-epitope on EEA2 was determined. Coiled-coil region 3 of EEA2 appeared to contain the epitope recognized by SW5. The SW5 epitope regions of La and EEA2 share a limited sequence homology and probably share a higher degree of structural similarity at the tertiary level. Most likely, the most critical determinants for recognition by SW5 reside in elements adopting alpha-helical conformations. These data indicate that the application of specific mAbs to purify and characterize (functionally) interacting proteins can be severely obscured by the cross-reactivity of mAbs with structurally, but not functionally, similar proteins.


Attenuated, oncolytic, but not wild-type measles virus infection has pleiotropic effects on human neutrophil function.
We previously showed that neutrophils play a role in regression of human tumor xenografts in immunodeficient mice following oncolytic vaccine measles virus (MV-Vac) treatment. In this study, we sought, using normal human neutrophils, to identify potential neutrophil-mediated mechanisms for the attenuated MV-Vac induced effects seen in vivo, by comparison with those consequent on wild-type (WT-MV) infection. Both MV-Vac and WT-MV infected and replicated within neutrophils, despite lack of SLAM expression. In both cases, neutrophils survived longer ex vivo postinfection. Furthermore, MV-Vac (but not WT-MV) infection activated neutrophils and stimulated secretion of several specific antitumor cytokines (IL-8, TNF-α, MCP-1, and IFN-α) via induction of de novo RNA and protein synthesis. In addition, MV-Vac (but not WT-MV) infection caused TRAIL secretion in the absence of de novo synthesis by triggering release of prefabricated TRAIL, via a direct effect upon degranulation. The differences between the outcome of infection by MV-Vac and WT-MV were not entirely explained by differential infection and replication of the viruses within neutrophils. To our knowledge, this is the first demonstration of potential mechanisms of oncolytic activity of an attenuated MV as compared with its WT parent. Furthermore, our study suggests that neutrophils have an important role to play in the antitumor effects of oncolytic MV.


Perforated duodenal diverticulitis: a report of three cases.
Duodenal diverticuli are present in up to 22% of the population. However, perforation of a duodenal diverticulum with spillage of enteric contents into the retroperitoneum is rare. We report three cases of perforated duodenal diverticulitis. Clinical presentations varied widely from patients with acute abdominal findings and generalized sepsis to a patient with mild symptoms of abdominal discomfort. CT scanning was the imaging modality used to make an accurate diagnosis. Treatment approaches for the most stable patient included nonoperative management with antibiotics, bowel rest and parenteral alimentation, while the less stable patients underwent definitive surgery with complete diversion of gastric contents and biliary flow from the affected area of duodenum. This report highlights the salient issues in the presentation, diagnosis and modern management of patients with this potentially catastrophic disease.


Comparative kinetic isotope effects on first- and second-order rate constants of soybean lipoxygenase variants uncover a substrate-binding network.
Lipoxygenases are widespread enzymes found in virtually all eukaryotes, including fungi, and, more recently, in prokaryotes. These enzymes act on long-chain polyunsaturated fatty acid substrates (C18 to C20), raising questions regarding how the substrate threads its way from solvent to the active site. Herein, we report a comparison of the temperature dependence of isotope effects on first- and second-order rate constants among single-site variants of the prototypic plant enzyme soybean lipoxygenase-1 substituted at amino acid residues inferred to impact substrate binding. We created 10 protein variants including four amino acid positions, Val-750, Ile-552, Ile-839, and Trp-500, located within a previously proposed substrate portal. The conversion of these bulky hydrophobic side chains to smaller side chains is concluded to increase the mobility of flanking helices, giving rise to increased off rates for substrate dissociation from the enzyme. In this manner, we identified a specific "binding network" that can regulate movement of the substrate from the solvent to the active site. Taken together with our previous findings on C-H and O2 activation of soybean lipoxygenase-1, these results support the emergence of multiple complementary networks within a single protein scaffold that modulate different steps along the enzymatic reaction coordinate.


Genetic characterization and phylogenetic analysis of rabbit hemorrhagic disease virus isolated in Tunisia from 2015 to 2018.
Two distinct genotypes responsible for rabbit hemorrhagic disease (RHD) are reported, GI.1 (RHDV) and GI.2 (RHDV2). Vaccines based on these two genotypes are only partially cross-protective. Hence, knowing which genotype is circulating is important for appropriate control measures. We have investigated 25 field samples isolated between 2015 and 2018 from rabbits with clinical signs of RHD. Only GI.2 (RHDV2) is currently circulating in Tunisia. All Tunisian samples were grouped together with typical genotypic and phenotypic mutations. Therefore, we recommend initiating an extensive preventive vaccination program based on GI.2 vaccines in addition to a regular monitoring of the circulating lagoviruses.


Statistical approaches to outcomes assessment.
Statistical methods and software can be useful for evaluating clinical outcomes data. The use of control charts and regression analysis can be particularly helpful in outcomes assessment. Control charts can reveal outlier events and patterns that require additional review leading to changes that improve outcomes. Regression analysis can show factors that affect process characteristics. This article uses examples derived from hospital outcomes assessment activities relating to length of stay, treatment planning, insurance coverage, patient characteristics, and clinical decision making. Minitab statistical software is used to create control charts and perform regression analysis, and essential Minitab commands are explained. Through the use of the techniques described the clinical and manager can more effectively evaluate clinical outcomes data to improve healthcare quality.


Modifying fatty acid profiles through a new cytokinin-based plastid transformation system.
The widespread use of herbicides and antibiotics for selection of transgenic plants has not been very successful with regard to commercialization and public acceptance. Hence, alternative selection systems are required. In this study, we describe the use of ipt, the bacterial gene encoding the enzyme isopentenyl transferase from Agrobacterium tumefaciens, as a positive selectable marker for plastid transformation. A comparison between the traditional spectinomycin-based aadA selection system and the ipt selection system demonstrated that selection of transplastomic plants on medium lacking cytokinin was as effective as selection on medium containing spectinomycin. Proof of principle was demonstrated by transformation of the kasIII gene encoding 3-ketoacyl acyl carrier protein synthase III into tobacco plastids. Transplastomic tobacco plants were readily obtained using the ipt selection system, and were phenotypically normal despite over-expression of isopentenyl transferase. Over-expression of KASIII resulted in a significant increase in 16:0 fatty acid levels, and a significant decrease in the levels of 18:0 and 18:1 fatty acids. Our study demonstrates use of a novel positive plastid transformation system that may be used for selection of transplastomic plants without affecting the expression of transgenes within the integrated vector cassette or the resulting activity of the encoded protein. This system has the potential to be applied to monocots, which are typically not amenable to traditional antibiotic-based selection systems, and may be used in combination with a negative selectable marker as part of a two-step selection system to obtain homoplasmic plant lines.


Genetic structure and origin of peopling in the Azores islands (Portugal): the view from mtDNA.
The Azores islands (Portugal), uninhabited when discovered by Portuguese navigators in the fifteenth century, are located in the Atlantic Ocean 1500 km from the European mainland. The archipelago is formed by nine islands of volcanic origin that define three geographical groups: Eastern (S. Miguel and Sta. Maria), Central (Terceira, Faial, Pico, Graciosa and S. Jorge) and Western (Flores and Corvo). To improve the genetic characterisation of the Azorean population, and to clarify some aspects related to the history of settlement, a study of mtDNA was conducted in the population of the archipelago. The HVRI region was sequenced and specific RFLPs were screened in 146 samples obtained from unrelated individuals with Azorean ancestry (50 from the Eastern group, 60 from the Central group, and 37 from the Western group). Samples were classified into haplogroups based on the information obtained from both sequencing and RFLP analysis. All the analyses performed support the idea that, in the whole group of islands, the majority of mtDNA lineages originated from the Iberian Peninsula, mainly from Portugal (mainland). However contributions from other European populations, especially from Northern Europe, cannot be disregarded. The values obtained for the various diversity parameters in the Azores archipelago indicate that the Azorean population, as a whole, does not exhibit the typical characteristics of an isolated population. The analysis of genetic data by groups of islands showed that the Western group exhibited particular features. The distribution of haplogroups in the Western group is very atypical, being significantly different from what is observed in the Eastern and Central groups. Furthermore, the diversity values are, in general, lower than those observed in other populations used for comparison. African haplogroups were found in all the groups of islands. Therefore the presence of Moorish and African slaves on the islands, as reported in historical sources, is supported by the mtDNA genetic data, especially in the Eastern group. The presence of Jews in the Central group is also supported by the mtDNA data. Neither historical nor genetic data (phylogeography of mtDNA) supports the idea of a differential settlement history for the Western group; however, it is represented in the phylogenies as an isolated branch. The effect of genetic drift, induced by the reduced population size since peopling occurred, has led to a very atypical distribution of haplogroups/haplotypes in this group of islands. We cannot ignore the influence of biodemographic and genetic processes, namely founder effect, genetic drift, migration, and even recent mutational events in the mtDNA lineages of the Azorean populations. Nevertheless, a great part of the variation in the Azorean mtDNA can be explained by the settlement history.


Concurrent validation of estimated activity energy expenditure using a 3-day diary and accelerometry in adolescents.
Estimates of daily energy expenditure are important to studies of physical activity and energy balance. Objective measures are not always feasible and further research is needed to validate survey instruments and diaries. The study validates estimated activity energy expenditure (AEE) based on a 3-day diary protocol relative to AEE derived from uniaxial accelerometry in adolescents, 265 girls and 227 boys (12.5-16.4 years). Participants completed the diary and wore a GT1M Actigraph accelerometer on the same days. Height and weight were measured. Correlations between protocols were significant (P<0.001) but moderate, r=0.65 in males and r=0.69 in females. The highest correlation occurred among males on Friday, r=0.74 (P<0.01). Controlling for body mass, partial correlations between protocols decreased to 0.44 and 0.35 in males and females, respectively. About 97% of the cases fell within the limits of agreement in a Bland-Altman plot. The criterion of inclusion for the accelerometer excluded 18% of the initial sample. In summary, the 3-day diary was completed without any major problems and provided a reasonably valid alternative for assessing AEE. Concordance between methods was slightly lower for individuals with higher values of AEE.


Rabphilin 3A binds the N-peptide of SNAP-25 to promote SNARE complex assembly in exocytosis.
Exocytosis of secretory vesicles requires the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins and small GTPase Rabs. As a Rab3/Rab27 effector protein on secretory vesicles, Rabphilin 3A was implicated to interact with SNAP-25 to regulate vesicle exocytosis in neurons and neuroendocrine cells, yet the underlying mechanism remains unclear. In this study, we have characterized the physiologically relevant binding sites between Rabphilin 3A and SNAP-25. We found that an intramolecular interplay between the N-terminal Rab-binding domain and C-terminal C2AB domain enables Rabphilin 3A to strongly bind the SNAP-25 N-peptide region via its C2B bottom α-helix. Disruption of this interaction significantly impaired docking and fusion of vesicles with the plasma membrane in rat PC12 cells. In addition, we found that this interaction allows Rabphilin 3A to accelerate SNARE complex assembly. Furthermore, we revealed that this interaction accelerates SNARE complex assembly via inducing a conformational switch from random coils to α-helical structure in the SNAP-25 SNARE motif. Altogether, our data suggest that the promotion of SNARE complex assembly by binding the C2B bottom α-helix of Rabphilin 3A to the N-peptide of SNAP-25 underlies a pre-fusion function of Rabphilin 3A in vesicle exocytosis.


The sucrose-regulated Arabidopsis transcription factor bZIP11 reprograms metabolism and regulates trehalose metabolism.
• The Arabidopsis basic region-leucine zipper transcription factor 11 (bZIP11) is known to be repressed by sucrose through a translational inhibition mechanism that requires the conserved sucrose control peptide encoded by the mRNA leader. The function of bZIP11 has been investigated in over-expression studies, and bZIP11 has been found to inhibit plant growth. The addition of sugar does not rescue the growth inhibition phenotype. Here, the function of the bZIP11 transcription factor was investigated. • The mechanism by which bZIP11 regulates growth was studied using large-scale and dedicated metabolic analysis, biochemical assays and molecular studies. • bZIP11 induction results in a reprogramming of metabolism and activation of genes involved in the metabolism of trehalose and other minor carbohydrates such as myo-inositol and raffinose. bZIP11 induction leads to reduced contents of the prominent growth regulatory molecule trehalose 6-phosphate (T6P). • The metabolic changes detected mimic in part those observed in carbon-starved plants. It is proposed that bZIP11 is a powerful regulator of carbohydrate metabolism that functions in a growth regulatory network that includes T6P and the sucrose non-fermenting-1 related protein kinase 1 (SnRK1).


Analysis of Clostridium difficile biofilms: imaging and antimicrobial treatment.
Clostridium difficile, a spore-forming Gram-positive anaerobic bacillus, is the most common causative agent of healthcare-associated diarrhoea. Formation of biofilms may protect C. difficile against antibiotics, potentially leading to treatment failure. Furthermore, bacterial spores or vegetative cells may linger in biofilms in the gut causing C. difficile infection recurrence. In this study, we evaluated and compared the efficacy of four antibiotics (fidaxomicin, surotomycin, vancomycin and metronidazole) in penetrating C. difficile biofilms and killing vegetative cells. C. difficile biofilms grown initially for 48 or 72 h using the colony biofilm model were then treated with antibiotics at a concentration of 25 × MIC for 24 h. Vegetative cells and spores were enumerated. The effect of treatment on biofilm structure was studied by scanning electron microscopy (SEM). The ability of fidaxomicin and surotomycin to penetrate biofilms was studied using fluorescently tagged antibiotics. Both surotomycin and fidaxomicin were significantly more effective than vancomycin or metronidazole (P < 0.001) at killing vegetative cells in established biofilms. Fidaxomicin was more effective than metronidazole at reducing viable spore counts in biofilms (P < 0.05). Fluorescently labelled surotomycin and fidaxomicin penetrated C. difficile biofilms in < 1 h. After 24 h of treatment, SEM demonstrated that both fidaxomicin and surotomycin disrupted the biofilm structure, while metronidazole had no observable effect. Fidaxomicin is effective in disrupting C. difficile biofilms, killing vegetative cells and decreasing spore counts.


Deficits in facial emotion recognition indicate behavioral changes and impaired self-awareness after moderate to severe traumatic brain injury.
Traumatic brain injury (TBI) is a leading cause of disability, specifically among younger adults. Behavioral changes are common after moderate to severe TBI and have adverse consequences for social and vocational functioning. It is hypothesized that deficits in social cognition, including facial affect recognition, might underlie these behavioral changes. Measurement of behavioral deficits is complicated, because the rating scales used rely on subjective judgement, often lack specificity and many patients provide unrealistically positive reports of their functioning due to impaired self-awareness. Accordingly, it is important to find performance based tests that allow objective and early identification of these problems. In the present study 51 moderate to severe TBI patients in the sub-acute and chronic stage were assessed with a test for emotion recognition (FEEST) and a questionnaire for behavioral problems (DEX) with a self and proxy rated version. Patients performed worse on the total score and on the negative emotion subscores of the FEEST than a matched group of 31 healthy controls. Patients also exhibited significantly more behavioral problems on both the DEX self and proxy rated version, but proxy ratings revealed more severe problems. No significant correlation was found between FEEST scores and DEX self ratings. However, impaired emotion recognition in the patients, and in particular of Sadness and Anger, was significantly correlated with behavioral problems as rated by proxies and with impaired self-awareness. This is the first study to find these associations, strengthening the proposed recognition of social signals as a condition for adequate social functioning. Hence, deficits in emotion recognition can be conceived as markers for behavioral problems and lack of insight in TBI patients. This finding is also of clinical importance since, unlike behavioral problems, emotion recognition can be objectively measured early after injury, allowing for early detection and treatment of these problems.


[Peripheral blood picture following dietary changes and neuro-emotional stress].
The effects of diets prepared for prolonged space flights and stress exposures on peripheral blood of 42 healthy male test subjects was studied. The morphology of peripheral blood remained essentially unaltered, when test subjects were given dehydrated diets (freshly prepared, stored for 2 years and exposed to proton irradiation of a dose of 24,000 rad) or Salyut-6 space diet. Stress effects brought about neutrophil leukocytosis and lymphopenia which could be prevented with dietary supplements.


H-2 antigen requirements in the in vitro induction of SV40-specific cytotoxic T lymphocytes.
T cytotoxic cells generated to syngeneic SV40 virus transformants lyse only SV40 target cells that are syngeneic at the H-2 locus. In contrast, SV40-specific tumor transplantation immunity shows no requirements for syngeneic H-2. Inoculation of allogeneic or even xenogeneic transformants will confer immunity to a challenge of syngeneic SV40 tumor cells. The experiments described here represent an attempt to reconcile these apparently conflicting observations. In our hands, generation of SV40-specific T cytotoxic cells in vitro requires both in vivo priming and secondary in vitro sensitization. We have found that priming for a secondary syngeneic-restricted response requires only that the cell employed be SV40 transformed. That is, priming may be accomplished with syngeneic, allogeneic, or xenogeneic SV40 transformants. Thus, the apparent lack of H-2 restriction in vivo immunity does not eliminate a role for the H-2-restricted cytotoxic T cell in tumor transplantation immunity.


Impact of surface roughness on the effective dielectric constants and subwavelength image resolution of metal-insulator stack lenses.
The effective parallel and perpendicular dielectric constants for a multilayer metal–insulator stack are obtained from numerical simulations and compared with analytical homogenization results as a function of wavelength and number of periods. The influence of inevitable film surface roughness on the homogenized dielectric constants, determined from numerical scattered field calculations, is evaluated as a function of roughness. The impact of this roughness on resolution in a subwavelength imaging application gives smoothness guidelines for material deposition.


Health literacy and anticoagulation-related outcomes among patients taking warfarin.
Little is known about whether health literacy affects anticoagulation-related outcomes. To assess how health literacy is associated with warfarin knowledge, adherence, and warfarin control (measured by the international normalized ratio [INR]). Survey. Patients taking warfarin through an anticoagulation clinic. Health literacy was measured using the short-form Test of Functional Health Literacy in Adults (s-TOFHLA), dichotomized as "limited" (score 0 to 22) and "adequate" (score 23 to 36). We asked patients to answer questions relating to their warfarin therapy and used multivariable logistic regression to assess whether health literacy was associated with incorrect answers. We also assessed whether health literacy was associated with nonadherence to warfarin as well as time in therapeutic INR range. Bilingual research assistants administered the survey and s-TOFHLA to 179 anticoagulated English- or Spanish-speaking patients. Limited health literacy was associated with incorrect answers to questions on warfarin's mechanism (adjusted odds ratio [OR] 4.8 [1.3 to 17.6]), side-effects (OR 6.4 [2.3 to 18.0]), medication interactions (OR 2.5 [1.1 to 5.5]), and frequency of monitoring (OR 2.7 [1.1 to 6.7]), after adjusting for age, sex, race/ethnicity, education, cognitive impairment, and years on warfarin. However, limited health literacy was not significantly associated with missing warfarin doses in 3 months (OR 0.9 [0.4 to 2.0]) nor with the proportion of person-time in therapeutic INR range (OR 1.0 [0.7 to 1.4]). Limited health literacy is associated with deficits in warfarin-related knowledge but not with self-reported adherence to warfarin or INR control. Efforts should concentrate on investigating alternative means of educating patients on the management and potential risks of anticoagulation.


Burkitt's lymphoma of the duodenum in a patient with AIDS.
Non-Hodgkin's lymphoma of B-cell type is the second most common neoplasm after Kaposi's sarcoma, among patients with human immunodeficiency virus infection. Most non-Hodgkin's lymphoma cases that are associated with acquired immunodeficiency syndrome involve extranodal sites, especially the digestive tract and the central nervous system. We report a case of primary lymphoma of the duodenum in a patient with AIDS. Upper gastrointestinal endoscopy revealed pseudopolypoid masses found in the second portion of the duodenum. A complete diagnostic study including histological, immunohistochemical and virological analyses showed high-grade B-cell Burkitt's lymphoma. The Epstein-Barr virus genome was detected in biopsies by immunohistochemical and in situ hybridization.


Patients' decision-making experiences in the acute healthcare setting--a case study.
The transition from being an ordinary citizen to a hospitalised patient can be a daunting experience particularly for the uninitiated and inexperienced. Patients are likely to have questions such as 'where do I go?', 'what should I do?', 'when?' and 'who should I ask?' The process for making practical moment-to-moment decisions is often complex and fraught with difficulties. Identifying critical points in the hospitalisation experience may provide insights into the quality of hospital management systems and professional practices from a patient perspective. This study aimed to identify institutional practices and structures in the context of acute healthcare settings which impact on patient moment-to-moment decision-making experiences. A case study approach was used as the exploratory methodology, and interviews were conducted with three former adult inpatients. In order to gain an understanding of each participant's experiences, data collecting strategies used in this research were a semistructured interview and document analysis of information documents, such as hospital supplied pamphlets, provided to the researcher by the participants. The study identified five major themes. Identified as being critical to the participants were information sharing by healthcare professionals, professional advice and professional role identification. Less so were environment and everyday life. Associated with these themes were the participant's common experience of being confined in unfamiliar surroundings, adjusting to institutional routines and of being heavily dependent on others. Findings indicate that patient moment-to-moment decision-making may be informed and enhanced in several areas: relevant and timely information sharing, varying the dress code between the different professions, reducing conflicting professional advice, clear signage around the hospital, and flexible visiting hours, telephones and clocks in patient rooms.


Associations between smoking and extra-axial manifestations and disease severity in axial spondyloarthritis: results from the BSR Biologics Register for Ankylosing Spondylitis (BSRBR-AS).
The effects of smoking on disease manifestations in axial SpA are inadequately described. Utilizing a large and well-characterized cohort, we investigated the association between smoking and extra-axial manifestations, and smoking and disease severity measures. Baseline data from the British Society for Rheumatology Biologics Register for Ankylosing Spondylitis were explored. Our analyses focused on extra-axial manifestations and other disease severity measures, including scales for fatigue, sleep, anxiety and depression. Logistic and linear models were used to quantify associations between disease characteristics according to smoking status (current/ex/never) and quantity (heavy/light), adjusting for age, gender, BMI, education, deprivation, comorbidities, symptom duration and alcohol status. A total of 2031 participants were eligible for the current analysis (68% male, mean age 49 years). Of these, 24% were current and 32% ex-smokers. When compared with non-smokers, current smokers had lower odds of uveitis [OR 0.7, 95% CI 0.5-0.9] and higher odds of psoriasis (ORadj 1.6, 95% CI 1.1-2.3). Ex- and current smokers had incrementally more severe disease than never smokers, with higher BASDAI (β = 0.3, 95% CI 0.1-0.6; β = 0.9, 95% CI 0.6-1.2) and BASFI (β = 0.5, 95% CI 0.2-0.8; β = 1.3, 95% CI 1.0-1.6); similar associations were observed for fatigue, sleep, anxiety and depression. In this large cross-sectional study, we observed that smoking is independently associated with an adverse disease profile in axial SpA, including worse fatigue, sleep, anxiety and depression, and higher odds of psoriasis. The paradoxical association between current smoking and reduced odds of uveitis is interesting and warrants further investigation.


Regulation of cellular and system function by activin.
Activin is an important molecule that regulates hormonogenesis, cellular homeostasis (divide or die pathways), and differentiation programs (developmentally and in adult cells). The cellular mechanisms that integrate an activin signal into a physiological response include a binary receptor complex and tandem serine threonine kinases, intracellular signal mediators, and nuclear transcription factors. Activin antagonists (inhibins) and bioneutralizing binding proteins (follistatins) act as gating molecules to ensure accurate delivery of activin signals to cellular machinery. Correct execution of an activin cue intracellularly permits actions as fundamental as embryonic mesoderm development, neuronal survival, hematopoietic function, and reproductive cyclicity. Absent or incorrect activin signaling results in phenotypes as catastrophic as embryonic lethality, tumor formation, and infertility. The general ways in which a cell senses and responds to an activin signal will be reviewed in the first part of this paper. The role of this ligand in reproductive function will also be examined as a specific example of activin activity.


Interfacial exchange coupling induced anomalous anisotropic magnetoresistance in epitaxial γ'-Fe₄N/CoN bilayers.
Anisotropic magnetoresistance (AMR) of the facing-target reactively sputtered epitaxial γ'-Fe4N/CoN bilayers is investigated. The phase shift and rectangular-like AMR appears at low temperatures, which can be ascribed to the interfacial exchange coupling. The phase shift comes from the exchange bias (EB) that makes the magnetization lag behind a small field. When the γ'-Fe4N thickness increases, the rectangular-like AMR appears. The rectangular-like AMR should be from the combined contributions including the EB-induced unidirectional anisotropy, intrinsic AMR of γ'-Fe4N layer and interfacial spin scattering.


Phenobarbital Induces SLC13A5 Expression through Activation of PXR but Not CAR in Human Primary Hepatocytes.
Phenobarbital (PB), a widely used antiepileptic drug, is known to upregulate the expression of numerous drug-metabolizing enzymes and transporters in the liver primarily via activation of the constitutive androstane receptor (CAR, NR1I3). The solute carrier family 13 member 5 (SLC13A5), a sodium-coupled citrate transporter, plays an important role in intracellular citrate homeostasis that is associated with a number of metabolic syndromes and neurological disorders. Here, we show that PB markedly elevates the expression of SLC13A5 through a pregnane X receptor (PXR)-dependent but CAR-independent signaling pathway. In human primary hepatocytes, the mRNA and protein expression of SLC13A5 was robustly induced by PB treatment, while genetic knockdown or pharmacological inhibition of PXR significantly attenuated this induction. Utilizing genetically modified HepaRG cells, we found that PB induces SLC13A5 expression in both wild type and CAR-knockout HepaRG cells, whereas such induction was fully abolished in the PXR-knockout HepaRG cells. Mechanistically, we identified and functionally characterized three enhancer modules located upstream from the transcription start site or introns of the SLC13A5 gene that are associated with the regulation of PXR-mediated SLC13A5 induction. Moreover, metformin, a deactivator of PXR, dramatically suppressed PB-mediated induction of hepatic SLC13A5 as well as its activation of the SLC13A5 luciferase reporter activity via PXR. Collectively, these data reveal PB as a potent inducer of SLC13A5 through the activation of PXR but not CAR in human primary hepatocytes.


Is there a dysfunction in the visual system of depressed patients?
BACKGROUND: The aim of the current study was to identify a possible locus of dysfunction in the visual system of depressed patients. MATERIALS AND METHODS: Fifty Major Depressive patients aged 21-60 years and 15 age-matched controls took part in the study The diagnosis was obtained with the SCAN v 2.0. The psychometric assessment included the HDRS, the HAS, the Newcastle Scales, the Diagnostic Melancholia Scale and the GAF scale. Flash Electroretinogram and Electrooculogram were performed in all subjects. The statistical analysis included ANCOVA, Student's t-test and Pearson Product Moment Correlation Coefficient were used. RESULTS: The Electro-oculographic findings suggested that all subtypes of depressed patients had lower dark trough and light peak values in comparison to controls (p < 0.001), while Arden ratios were within normal range. Electroretinographic recordings did not reveal any differences between patients and controls or between subtypes of depression. DISCUSSION: The findings of the current study provide empirical data in order to assist in the understanding of the international literature and to explain the mechanism of action of therapies like sleep deprivation and light therapy.


Demonstration of collagen type VI and alpha-smooth muscle actin in renal fibrotic injury in man.
Overproduction of collagenous fibres types I and III is a common finding of fibrotic injury. Collagen type VI is generally associated with type I. Appearance of fibroblasts expressing alpha-smooth muscle actin (ASMA) and their role in fibrogenesis has been partly defined. However, correlation between renal fibroblasts and accumulation of microfibrillar collagen type VI, as well as its exact distribution, is not fully delineated. This study was undertaken to investigate these issues using a complex morphological approach. Morphological examination included immunohistochemical detection of the collagen type VI and ASMA, relying on a streptavidin-biotin-peroxidase-based technique, and electron microscopy. Collagen type VI was strongly expressed in areas of fibrotic injury, although mild expression was always revealed in renal interstitium. Glomerular immunoreactivity with the anti-collagen type VI antibody was almost nil excepting cases of diabetic glomerulosclerosis and amyloid nephrosis. Glomerular nodules in cases of diabetes displayed intense reactivity. Mesangial, as well as discontinuous peripheral deposition of collagen along the glomerular basement membrane, was noticed in case of amyloidosis. Ultrastructurally, cross-banded collagen microfibrils were found in renal interstitium in close association with the fibroblast membrane. Moreover, fibrillar elements revealing tubular structure and fine filamentous material were observed between cross-banded microfibrils. Some of fibroblasts exhibited bundles of microfilaments in their cytoplasm. An increased number of ASMA-positive cells was detected in fibrotic interstitium. An intense concentric network made up of actin-bearing cells surrounded glomerular capillaries in the case of crescentic glomerular lesions. Markedly increased deposition of collagen type VI takes place in renal fibrotic lesions. Simultaneously, interstitial fibrotic areas appeared to contain a great number of fibroblasts sharing morphological characteristics of classic fibroblasts and smooth muscle cells. Detailed examination of coexistence of these two interstitial phenomena should further clarify the cellular mechanisms involved in renal interstitial fibrosis.


Comparative Transcriptional Profiling of Two Contrasting Barley Genotypes under Salinity Stress during the Seedling Stage.
Salinity is one of the major abiotic stresses that affect crop productivity. Identification of the potential novel genes responsible for salt tolerance in barley will contribute to understanding the molecular mechanism of barley responses to salt stress. We compared changes in transcriptome between Hua 11 (a salt-tolerant genotype) and Hua 30 (a salt sensitive genotype) in response to salt stress at the seedling stage using barley cDNA microarrays. In total, 557 and 247 salt-responsive genes were expressed exclusively in the shoot and root tissue of the salt-tolerant genotype, respectively. Among these genes, a number of signal-related genes, transcription factors and compatible solutes were identified and some of these genes were carefully discussed. Notably, a LysM RLK was firstly found involved in salt stress response. Moreover, key enzymes in the pathways of jasmonic acid biosynthesis, lipid metabolism and indole-3-acetic acid homeostasis were specifically affected by salt stress in salt tolerance genotype. These salt-responsive genes and biochemical pathways identified in this study could provide further information for understanding the mechanisms of salt tolerance in barley.


POSSIBLE SOURCE OF SECONDARY INVADING STAPHYLOCOCCI IN MICE EXPOSED TO ACUTE COLD.
Miraglia, Gennaro J. (Bryn Mawr College, Bryn Mawr, Pa.) and L. Joe Berry. Possible source of secondary invading staphylococci in mice exposed to acute cold. J. Bacteriol. 85:345-348. 1963.-In an effort to determine the origin of the staphylococci known to invade the deep tissues (liver, spleen, kidneys, lungs, and heart) of mice exposed continuously to 5 C in individual compartments without bedding, the intestinal tract was freed of these organisms, as judged by absence of growth when fecal suspensions were inoculated into selective media. Substitution of 0.01 n hydrochloric acid for drinking water eliminated staphylococci within a few days, yet the incidence of tissue invasion was unaltered. The coagulase-negative strains normally present in feces and in tissues persisted in tissues even though the intestine was seeded with a coagulase-positive strain by feeding contaminated food. Cultures from the external nares continued unaltered. This suggests that the respiratory tract is a possible origin of the staphylococci found in tissues of the cold-stressed mice.


Intracranial arachnoid cysts in children.
The clinical and radiographic findings, surgical treatment, and outcome in 16 pediatric patients with intracranial arachnoid cysts are reviewed. The clinical presentation reflected the anatomical location of the lesions. Computerized tomography or magnetic resonance imaging scans were diagnostic in all cases. Of the nine cysts treated primarily or secondarily by craniotomy for fenestration and drainage into the basilar cisterns, five recurred. Cyst-peritoneal shunting led to diminished cyst size and clinical improvement in all seven cases in which it was used as the initial treatment and in all four cases in which fenestration had been unsuccessful. The results in this series show that cyst-peritoneal shunting is the treatment of choice for most intracranial arachnoid cysts in children.


Oxygen-regulated post-translation modifications as master signalling pathway in cells.
Oxygen is essential for viability in mammalian organisms. However, cells are often exposed to changes in oxygen availability, due to either increased demand or reduced oxygen supply, herein called hypoxia. To be able to survive and/or adapt to hypoxia, cells activate a variety of signalling cascades resulting in changes to chromatin, gene expression, metabolism and viability. Cellular signalling is often mediated via post-translational modifications (PTMs), and this is no different in response to hypoxia. Many enzymes require oxygen for their activity and oxygen can directly influence several PTMS. Here, we review the direct impact of changes in oxygen availability on PTMs such as proline, asparagine, histidine and lysine hydroxylation, lysine and arginine methylation and cysteine dioxygenation, with a focus on mammalian systems. In addition, indirect hypoxia-dependent effects on phosphorylation, ubiquitination and sumoylation will also be discussed. Direct and indirect oxygen-regulated changes to PTMs are coordinated to achieve the cell's ultimate response to hypoxia. However, specific oxygen sensitivity and the functional relevance of some of the identified PTMs still require significant research.


Pain resilience moderates the influence of negative pain beliefs on movement-evoked pain in older adults.
Negative pain beliefs are associated with adverse pain outcomes; however, less is known regarding how positive, adaptive factors influence pain and functioning. These relationships are especially important to examine in older adults with pain, given increased disability and functional limitations in this population. We investigated whether pain resilience moderated the relationships between negative pain beliefs (fear-avoidance, pain catastrophizing) and pain outcomes (functional performance, movement-evoked pain) in sixty older adults with low back pain. Higher pain resilience was associated with lower fear-avoidance (p < .05) and pain catastrophizing (p = .05). After controlling for demographic variables, higher fear-avoidance (p = .03) and catastrophizing (p = .03) were associated with greater movement-evoked pain in individuals with low pain resilience, but not among those high in resilience. No significant moderation effects were observed for functional performance. Resilience may attenuate the relationship between negative psychological processes and pain-related disability, highlighting the need for interventions that enhance pain resilience in older adults.


Computer-aided pattern analysis of temperature differentials.
Chiropractic analysis often incorporates the evaluation of heat distribution patterns observed in skin temperature profiles. Historically, temperature recordings of the spine have been made with a heat detection instrument using a thermocouple design. The advent of refinements in infrared technology, however, have greatly enhanced temperature detection in terms of accuracy and reliability. The present research reflects the development of computer software designed to express the data obtained with a dual channel heat sensing instrument. Two phases are involved: a) digital information, received from the sensing instrument, is standardized, stored for future analysis, and retrieved for comparisons with other graphs; b) stored data is graphically displayed, statistically analyzed, and otherwise compared. The data is displayed for visual observation as a) temperature data received from either channel of the sensing instrument or b) relative temperature differences between data from both channels of the sending instrument. For statistical evaluation, corresponding readings (comparisons between graphs) are plotted against one another and analyzed by a moving Pearson Product Moment correlation and moving t-test. This evaluation is graphically represented with a numeric display of pertinent statistical values. Current work indicates that a 10 point moving correlation and t-test will yield accurate comparisons between graphs.


Perinuclear location and recycling of epidermal growth factor receptor kinase: immunofluorescent visualization using antibodies directed to kinase and extracellular domains.
This paper describes studies on the migratory behavior of epidermal growth factor (EGF) receptor kinase using antibodies that are specific for either the kinase domain or the extracellular domain of the receptor. Antiserum was raised to a 42,000-D subfragment of EGF receptor, which was shown earlier to carry the kinase catalytic site but not the EGF-binding site. Another antiserum was raised to the pure intact 170,000-D EGF receptor. The specificities of these antibodies were established by immunoprecipitation and immunoblotting experiments. The domain specificity was examined by indirect immunofluorescent staining of fixed cells. The anti-42-kD peptide antibody could bind specifically to EGF receptors of both human and murine origin and was found to be directed to the cytoplasmic part of the molecule. It did not bind to EGF receptor-negative cells, which contained other types of tyrosine kinases. The antibodies raised against the intact receptor recognized only EGF receptor-specific epitopes and were directed to the extracellular part of the molecule. The anti-receptor antibodies described above were used to visualize the cyclic locomotory behavior of EGF receptor kinase under various conditions of EGF stimulation and withdrawal. The receptor was examined in fixed and permeabilized cells by indirect immunofluorescent staining. The results demonstrate the following: (a) the receptor kinase domain migrates to the perinuclear region upon challenge with EGF; (b) both extracellular and cytoplasmic domains of the receptor are involved in migration as a unit; (c) withdrawal of EGF results in rapid recycling of the perinuclear receptors to the plasma membrane; (d) this return to the cell surface is inhibited by methylamine, chloroquine, and monensin; and (e) neither the internal migration nor the recycling process is blocked by inhibitors of protein biosynthesis.


Antibodies and activity measurements for the detection of O-GlcNAc transferase and assay of its substrate, UDP-GlcNAc.
Since the discovery of O-GlcNAc modification (O-GlcNAcylation) 20 years ago, much attention has been given to OGT (O-GlcNAc transferase), the unique enzyme responsible for the nuclear and cytosolic O-GlcNAcylation processes. This review focuses on protocols that are routinely used to analyze OGT expression and activity. First are detailed techniques using rabbit polyclonal anti-OGT antibodies, namely, Western blot, (co-)immunoprecipitation, and immunofluorescence. We also describe the measurement of OGT activity by using synthetic peptides as acceptors and radiolabeled UDP-GlcNAc. Finally, a sensitive HPAEC-based technique to measure the cellular content of UDP-GlcNAc, the donor substrate of OGT, is described in detail.