Distinguishing Different Stages of Parkinson's Disease Using Composite Index of Speed and Pen-Pressure of Sketching a Spiral.
The speed and pen-pressure while sketching a spiral are lower among Parkinson's disease (PD) patients with higher severity of the disease. However, the correlation between these features and the severity level (SL) of PD has been reported to be 0.4. There is a need for identifying parameters with a stronger correlation for considering this for accurate diagnosis of the disease. This study has proposed the use of the Composite Index of Speed and Pen-pressure (CISP) of sketching as a feature for analyzing the severity of PD. A total of 28 control group (CG) and 27 PD patients (total 55 participants) were recruited and assessed for Unified Parkinson's Disease Rating Scale (UPDRS). They drew guided Archimedean spiral on an A3 sheet. Speed, pen-pressure, and CISP were computed and analyzed to obtain their correlation with severity of the disease. The correlation of speed, pen-pressure, and CISP with the severity of PD was -0.415, -0.584, and -0.641, respectively. Mann-Whitney U test confirmed that CISP was suitable to distinguish between PD and CG, while non-parametric k-sample Kruskal-Wallis test confirmed that it was significantly different for PD SL-1 and PD SL-3. This shows that CISP during spiral sketching may be used to differentiate between CG and PD and between PD SL-1 and PD SL-3 but not SL-2.


Assessing cardiometabolic parameter monitoring in inpatients taking a second-generation antipsychotic: The CAMI-SGA study - a cross-sectional study.
This study aims to determine the proportion of initial cardiometabolic assessment and its predicting factors in adults with schizophrenia, bipolar disorder or other related diagnoses for whom a second-generation antipsychotic was prescribed in the hospital setting. Cross-sectional study. The psychiatry unit of a Canadian tertiary care teaching hospital in Montreal, Canada. 402 patients with aforementioned disorders who initiated, restarted or switched to one of the following antipsychotics: clozapine, olanzapine, risperidone, paliperidone or quetiapine, between 2013 and 2016. We assessed the proportion of cardiometabolic parameters monitored. We identified predictors that influence the monitoring of cardiometabolic parameters and we assessed the proportion of adequate interventions following the screening of uncontrolled blood pressure and fasting glucose or glycated haemoglobin (HbA1c) results. Only 37.3% of patients received monitoring for at least three cardiometabolic parameters. Blood pressure was assessed in 99.8% of patients; lipid profile in 24.4%; fasting glucose or HbA1c in 33.3% and weight or body mass index in 97.8% of patients while waist circumference was assessed in 4.5% of patients. For patients with abnormal blood pressure and glycaemic values, 42.3% and 41.2% subsequent interventions were done, respectively. The study highlighted the psychiatric diagnosis (substance induced disorder OR 0.06 95% CI 0.00 to 0.44), the presence of a court-ordered treatment (OR 0.79 95% CI 0.35 to 1.79) and the treating psychiatrist (up to OR 34.0 95% CI 16.2 to 140.7) as predictors of cardiometabolic monitoring. This study reports suboptimal baseline cardiometabolic monitoring of patients taking an antipsychotic in a Canadian hospital. Optimising collaboration within a multidisciplinary team may increase cardiometabolic monitoring.


A biolistic-based genetic transformation system applicable to a broad-range of sugarcane and energycane varieties.
Sugarcane and energycane (Saccharum spp. hybrids) are prominent sources of sugar, ethanol, as well as high-value bioproducts globally. Genetic analysis for trait improvement of sugarcane is greatly hindered by its complex genome, limited germplasm resources, long breeding cycle, as well as recalcitrance to genetic transformation. Here, we present a biolistic-based transformation and bioreactor-based micro-propagation system that has been utilized successfully to transform twelve elite cane genotypes, yielding transformation efficiencies of up to 39%. The system relies on the generation of embryogenic callus from sugarcane and energycane apical shoot tissue, followed by DNA bombardment of embryogenic leaf roll discs (approximately one week) or calli (approximately 4 weeks). We present optimal criteria and practices for selection and regeneration of independent transgenic lines, molecular characterization, as well as a bioreactor-based micro-propagation technique, which can aid in rapid multiplication and analysis of transgenic lines. The cane transformation and micro-propagation system described here, although built on our previous protocols, has significantly accelerated the process of producing and multiplying transgenic material, and it is applicable to other varieties. The system is highly reproducible and has been successfully used to engineer multiple commercial sugarcane and energycane varieties. It will benefit worldwide researchers interested in genomics and genetics of sugarcane photosynthesis, cell wall, and bioenergy related traits.


Antidepressant activity of brahmi in albino mice.
In traditional system of medicine brahmi has been used to enhance memory. Recently it has been reported to have action in psychiatric disorders. With these backgrounds the work has been undertaken to study antidepressant activity of brahmi in albino mice. To evaluate antidepressant activity of brahmi in experimental models. The antidepressant activity was studied in albino mice using forced swimming test (FST), tail suspension test (TST) and shock induced depression (SID). Imipramine (10mg/kg), fluoxetine (30mg/kg) were used as standard drugs and brahmi (10, 20, 30mg/kg) was used as test drug. Brahmi exhibited significant decrease in duration of immobility in FST and reduced the shock induced decrease in activity in SID models. It didn't show any activity in the TST model. Brahmi has shown antidepressant activity in FST and SID.


Glycopeptide hormone production by cultured human diploid fibroblasts.
Human diploid fibroblasts in culture were examined for production of glycopeptide hormones. Forty-one percent of the strains produced human chorionic gonadotropin (hCG) under normal growth conditions. Constitutive hCG synthesis was apparently unrelated to donor age, length of time in culture, or number of passages. Follicle stimulating hormone (FSH) was not found in any strain investigated. Only one cell strain produced free alpha-chains of glycopeptide hormones. Hydroxyurea (HU) at a concentration of 1 mM mediated a small, statistically significant increase in hCG production (p less than 0.01) in all constitutive strains, but had no effect on non-hCG-producing fibroblast strains. Sodium butyrate (Bu) was effective in increasing hCG synthesis in only one constitutive strain, derived from a newborn foreskin. HU treatment had no apparent effect on cell structure. All Bu-treated strains, both those producing hCG and the nonproducers, showed morphological alterations; cells were flattened and they contained ordered arrays of refractile granules. It is suggested that hCG synthesis in cultured human diploid fibroblasts may result from a localized chromosomal event in which the loci responsible for this hormone are activated. Human diploid fibroblasts in culture are shown to be amenable to the study of gene expression and its modulation.


An exploration of site effects in a multisite trial of OROS-methylphenidate for smokers with attention deficit/hyperactivity disorder.
Multisite trials, the gold standard for conducting studies in community-based settings, can mask variability across sites resulting in misrepresentation of effects in specific sites. In a placebo-controlled trial of osmotic-release oral system methylphenidate (OROS-MPH) as augmentation treatment for smokers with attention deficit hyperactivity/impulsivity disorder (ADHD), three types of sites were selected according to their clinical research specialty (ADHD, smoking cessation, and general mental health). Analysis was conducted to determine if clinical outcomes, that is, reduction in ADHD symptoms and smoking cessation rates, and the effect of treatment on these outcomes would differ by type of site. A total of 255 adult smokers diagnosed with ADHD were enrolled in three clinic types: 72 in ADHD, 79 in tobacco dependence, and 104 in the mental health clinics. The three site-types were similar in demographic characteristics, smoking history, baseline level of ADHD symptoms, and history of psychiatric illness. Site-type but not a site-type by treatment interaction predicted prolonged smoking abstinence. A significant three-way interaction of site-type, treatment, and time-predicted improvement in ADHD symptoms. Moderate to strong effects of OROS-MPH relative to placebo were observed in the mental health and the ADHD clinics; a weak effect was observed in the tobacco dependence clinics. OROS-MPH benefit varied by site for reducing ADHD symptoms but not for improving smoking abstinence. Assessment of site-type effects can indicate the generalizability of findings from multisite trials and should be routinely incorporated in the design of multisite trials.


Association Between Skeletal Muscle Mass, Physical Activity, and Metabolic Syndrome: the Korean National Health and Nutrition Examination Survey 2008-2011.
Background: Skeletal muscle mass (SM) and physical activity (PA) are major modifiable factors that can moderate and prevent the occurrence of metabolic syndrome (MetS). However, the joint association between SM and PA guidelines in MetS remains unclear. Therefore, we aimed to examine the relationship between SM and PA with MetS among Korean adults. Methods: This cross-sectional study analyzed 18,090 adults from the Korean National Health and Nutrition Examination Survey 2008-2011. We used the value of appendicular skeletal muscle mass divided by body mass index as SM. We decided on the PA guidelines using the American College of Sports Medicine guidelines. After adjusting for confounding factors, we performed logistic regression analysis to calculate the odds ratio and 95% confidence interval of MetS associated with SM and PA guidelines. Results: Participants from the highest SM quartile showed a decreased MetS risk of 58%-75%. Those who met both aerobic and resistance exercise guidelines were more likely to have lower MetS risk than those who neither. In addition, even with the same PA guideline status, participants with the highest muscle mass decreased MetS risk by 29%-81% compared with participants with the lowest muscle mass. Conclusions: Our results showed that increased SM and meeting PA guidelines are significantly associated with a decreased risk of MetS. To prevent MetS, customized strategies are needed for improving muscle mass and PA according to age and gender.


Independent origin of plasmodium falciparum antifolate super-resistance, Uganda, Tanzania, and Ethiopia.
Super-resistant Plasmodium falciparum threatens the effectiveness of sulfadoxine-pyrimethamine in intermittent preventive treatment for malaria during pregnancy. It is characterized by the A581G Pfdhps mutation on a background of the double-mutant Pfdhps and the triple-mutant Pfdhfr. Using samples collected during 2004-2008, we investigated the evolutionary origin of the A581G mutation by characterizing microsatellite diversity flanking Pfdhps triple-mutant (437G+540E+581G) alleles from 3 locations in eastern Africa and comparing it with double-mutant (437G+540E) alleles from the same area. In Ethiopia, both alleles derived from 1 lineage that was distinct from those in Uganda and Tanzania. Uganda and Tanzania triple mutants derived from the previously characterized southeastern Africa double-mutant lineage. The A581G mutation has occurred multiple times on local Pfdhps double-mutant backgrounds; however, a novel microsatellite allele incorporated into the Tanzania lineage since 2004 illustrates the local expansion of emergent triple-mutant lineages.


[Validity and reproducibility of a physical activity questionnaire for adolescents: adapting the Self-Administered Physical Activity Checklist].
To evaluate test-retest reproducibility and the validity of a physical activity questionnaire for adolescents (adapted from Self-Administered Physical Activity Checklist). The test-retest reproducibility analysis was completed by 239 adolescents (average age 16 years, SD = 1.2; 56.4% female), and the validity analysis by 70 adolescents (average age 15.7 years, SD = 1.2; 55.7% female). All adolescents were high school students in João Pessoa, PB, in Northeastern Brazil, in 2009. The test-retest reproducibility was determined by repeated evaluations with one week interval between them. Validity of the physical activity questionnaire was assessed by comparing the results of the questionnaire with those of four 24-hour recalls (reference method). The test-retest reproducibility was high (intraclass correlation coefficient: ICC = 0.88; 95%CI: 0.84 - 0.91), and the agreement was moderate for physical activity classified in two groups (< 300 min/week vs. > 300 min/week; kappa coefficient: k = 0.52). Spearman correlation coefficient comparing the questionnaire data with those of four 24-hour recalls (reference method) was moderate (rho = 0.62; p < 0001). The agreement between these measures (questionnaire and 24-hours recalls) for physical activity classified in two groups was moderate (k = 0.59). The physical activity questionnaire showed high reproducibility, and moderate validity, and therefore, its use is recommended for assessing adolescents' physical activity level.


Genome-Wide Association Study Reveals Key Genes for Differential Lead Accumulation and Tolerance in Natural Arabidopsis thaliana Accessions.
Soil contamination by lead (Pb) has become one of the major ecological threats to the environment. Understanding the mechanisms of Pb transport and deposition in plants is of great importance to achieve a global Pb reduction. We exposed a collection of 360 Arabidopsis thaliana natural accessions to a Pb-polluted soil. Germination rates, growth, and leaf Pb concentrations showed extensive variation among accessions. These phenotypic data were subjected to genome wide association studies (GWAs) and we found a significant association on chromosome 1 for low leaf Pb accumulation. Genes associated with significant SNP markers were evaluated and we selected EXTENSIN18 (EXT18) and TLC (TRAM-LAG1-CLN8) as candidates for having a role in Pb homeostasis. Six Pb-tolerant accessions, three of them exhibiting low leaf Pb content, and three of them with high leaf Pb content; two Pb-sensitive accessions; two knockout T-DNA lines of GWAs candidate genes (ext18, tlc); and Col-0 were screened under control and high-Pb conditions. The relative expression of EXT18, TLC, and other genes described for being involved in Pb tolerance was also evaluated. Analysis of Darwinian fitness, root and leaf ionome, and TEM images revealed that Pb-tolerant accessions employ two opposing strategies: (1) low translocation of Pb and its accumulation into root cell walls and vacuoles, or (2) high translocation of Pb and its efflux to inactive organelles or intracellular spaces. Plants using the first strategy exhibited higher expression of EXT18 and HMA3, thicker root cell walls and Pb vacuolar sequestration, suggesting that these genes may contribute to the deposition of Pb in the roots. On the other hand, plants translocating high amounts of Pb showed upregulation of TLC and ABC transporters, indicating that these plants were able to properly efflux Pb in the aerial tissues. We conclude that EXT18 and TLC upregulation enhances Pb tolerance promoting its sequestration: EXT18 favors the thickening of the cell walls improving Pb accumulation in roots and decreasing its toxicity, while TLC facilitates the formation of dictyosome vesicles and the Pb encapsulation in leaves. These findings are relevant for the design of phytoremediation strategies and environment restoration.


Radiation inactivation of multimeric enzymes: application to subunit interactions of adenylate cyclase.
Radiation inactivation has been applied extensively to determine the molecular weight of soluble enzyme and receptor systems from the slope of a linear ln (activity) vs. dose curve. Complex nonlinear inactivation curves are predicted for multimeric enzyme systems, composed of distinct subunits in equilibrium with multimeric complexes. For the system A1 + A2----A1A2, with an active A1A2 complex (associative model), the ln (activity) vs. dose curve is linear for high dissociation constant, K. If a monomer, A1, has all the enzyme activity (dissociative model), the ln (activity) vs. dose curve has an activation hump at low radiation dose if the inactive subunit, A2, has a higher molecular weight than A1 and has upward concavity when A2 is smaller than A1. In general, a radiation inactivation model for a multistep mechanism for enzyme activation fulfills the characteristics of an associative or dissociative model if the reaction step forming active enzyme is an associative or dissociative reaction. Target theory gives the molecular weight of the active enzyme subunit or complex from the limiting slope of the ln (activity) vs. dose curve at high radiation dose. If energy transfer occurs among subunits in the multimer, the ln (activity) vs. dose curve is linear for a single active component and is concave upward for two or more active components. The use of radiation inactivation as a method to determine enzyme size and multimeric subunit assembly is discussed with specific application to the hormone-sensitive adenylate cyclase system. It is shown that the complex inactivation curves presented in the accompanying paper can be used select the best mechanism out of a series of seven proposed mechanisms for the activation of adenylate cyclase by hormone.


Spectroscopic approaches to elucidating novel iron-sulfur chemistry in the "radical-Sam" protein superfamily.
Electron paramagnetic resonance (EPR), electron-nuclear double resonance (ENDOR), and Mössbauer spectroscopies and other physical methods have provided important new insights into the radical-SAM superfamily of proteins, which use iron-sulfur clusters and S-adenosylmethionine to initiate H atom abstraction reactions. This remarkable chemistry involves the generation of the extremely reactive 5'-deoxyadenosyl radical, the same radical intermediate utilized in B12-dependent reactions. Although early speculation focused on the possibility of an organometallic intermediate in radical-SAM reactions, current evidence points to novel chemistry involving a site-differentiated [4Fe-4S] cluster. The focus of this forum article is on one member of the radical-SAM superfamily, pyruvate formate-lyase activating enzyme, and how physical methods, primarily EPR and ENDOR spectroscopies, are contributing to our understanding of its structure and mechanism. New ENDOR data supporting coordination of the methionine moiety of SAM to the unique site of the [4Fe-4S]2+/+ cluster are presented.


Children hospitalized for varicella: a prevaccine review.
To describe varicella complications in healthy and previously ill children hospitalized for varicella and to explore trends in group A beta-hemolytic streptococcus complications of varicella. A retrospective record review of children hospitalized for varicella between January 1, 1990, and March 31, 1994, was conducted in nine large acute care hospitals in Los Angeles County, California. We identified 574 children hospitalized for varicella in study hospitals during the 4.25-year study period (estimated risk of hospitalization, approximately 1 in 550 cases of varicella); 53% of the children were healthy before the onset of varicella and 47% were previously ill with underlying cancers or other chronic illnesses. Children were hospitalized for treatment of complications (n = 427, 74%) or for prophylactic antiviral therapy or observation (n = 147, 26%). Systems involved in complications included skin/soft tissue (45%), neurologic (18%), respiratory (14%), gastrointestinal (10%), and hematologic, renal, or hepatic (8% or less). The mean age of children with skin/soft tissue infections was 2.7 years (range < 1 to 16 years) compared with 4.7 years (< 1 to 18 years) for other complications. Children with skin/soft tissue and neurologic complications were more often previously healthy (p < 0.05), whereas those with respiratory complications were more often previously ill (p < 0.001). Hospitalizations for skin/soft tissue infections increased during the study period. The proportion of complications as a result of group A beta-hemolytic streptococcus infection increased from 4.7% before 1993 to 12.2% for the remainder of the study period (p = 0.02). Prior health status was predictive of the type of complications experienced by children with varicella requiring hospitalization. Our data suggest a recent increase in skin/soft tissue complications of varicella requiring hospitalization and an increase in the proportion of complications related to group A beta-hemolytic streptococcus. Wide-scale vaccine use should reverse this trend and reduce the overall impact of varicella on both healthy and previously ill children.


Rapid resolution of suicidal ideation after a single infusion of an N-methyl-D-aspartate antagonist in patients with treatment-resistant major depressive disorder.
Suicidal ideation is a medical emergency, especially when severe. Little research has been done on pharmacologic interventions that could address this problem. Ketamine, an N-methyl-D-asparate antagonist, has been reported to have antidepressant effects within hours. We examined the effects of a single dose of ketamine on suicidal ideation in subjects with treatment-resistant major depressive disorder (MDD). Thirty-three subjects with DSM-IV-diagnosed MDD received a single open-label infusion of ketamine (0.5 mg/kg) and were rated at baseline and at 40, 80, 120, and 230 minutes postinfusion with the Scale for Suicide Ideation (SSI), the Montgomery-Åsberg Depression Rating Scale, the Hamilton Depression Rating Scale, and the Beck Depression Inventory. The study was conducted between October 2006 and January 2009. Suicidal ideation scores decreased significantly on the SSI as well as on the suicide subscales of other rating instruments within 40 minutes; these decreases remained significant through the first 4 hours postinfusion (P < .001). Ten subjects (30%) had an SSI score ≥ 4 at baseline; all these scores dropped below 4 (9 dropped by 40 minutes and 1 by 80 minutes). For those patients with a starting score below 4 on the SSI, only 1 reached a score of 4. Depression, anxiety, and hopelessness were significantly improved at all time points (P < .001). Suicidal ideation in the context of MDD improved within 40 minutes of a ketamine infusion and remained improved for up to 4 hours postinfusion. Future studies with ketamine in suicidal ideation are warranted due to the potential impact on public health. clinicaltrials.gov Identifier: NCT00088699.


The preventive and therapeutic effect of acupuncture for radiation-induced xerostomia in patients with head and neck cancer: a systematic review.
Methods currently available to prevent and manage xerostomia which caused by radiotherapy in patients with head and neck caner have limited efficacy. Some studies suggest that acupuncture may be beneficial. The authors evaluated the preventive and therapeutic effect of acupuncture for radiation-induced xerostomia among patients with head and neck cancer. PUBMED, EMBASE, Cochrane Library, CBM, CAJD, Wan Fang database, and VIP Database for Chinese Technical Periodicals were electronically searched, in conjunction with further manual search for relevant articles. Studies that met the inclusion criteria were systematically evaluated. Three randomized controlled trials (RCTs) investigating the therapeutic effect of acupuncture were included. One RCT on the preventive effect of acupuncture was found. Because of the considerable variation among included studies, meta-analysis was not possible. Two included RCTs used placebo controls, and both observed significant improvement in the salivary flow rates between acupuncture and control groups. However, no significant differences were found. Three included RCTs suggested that acupuncture for radiation-induced xerostomia can improve patients' subjective symptoms. The only study evaluating the preventive effect of acupuncture for radiation-induced xerostomia showed positive changes in salivary flow rates (both unstimulated and stimulated) and dry mouth -related symptoms. Acupuncture treatment was well tolerated by all patients and no severe adverse effects were seen. Insufficient evidence is available to judge whether acupuncture is safe and whether it is effective in preventing or treating radiation-induced xerostomia. Significant research remains to be done before acupuncture can be recommended for routine use in radiation-induced xerostomia.


[Is lipid peroxidation responsible for the damage caused by postischemic reperfusion?].
Peroxidation of membrane phospholipid polyunsaturated fatty acids is considered a major mechanism of the damage occurring during post-ischemic reperfusion. The evidences in support for this mechanism of damage are based on tissue malondialdehyde (MDA) quantitation by the thiobarbituric acid test (TBA-test). In an attempt to verify this topic we have subjected isolated and Langendorff perfused rabbit hearts to a period of 60 min of severe ischemia plus 30 min of reperfusion. At appropriate time points MDA was determined in the tissue by means of TBA-test and directly by reversed phase, high pressure, liquid chromatography (HPLC). We have found no correlation between the 2 compared assays. During reperfusion, there was the formation of non-lipid related, MDA like, TBA-reactive substance which leads to overestimation of the extent of lipid peroxidation. On the contrary, by direct HPLC quantitation, there was a decrease of tissue MDA during ischemia and during the early phases of reperfusion. Our results demonstrate that TBA-test is not a reliable index of lipid peroxidation in organ systems and that MDA accumulation does not precede the evidence of the functional alterations occurring on reperfusion of the previously ischemic myocardium. These results are of relevance in the understanding of the exact mechanism of reperfusion damage as, in the same experimental model, oxy radicals have been shown to be generated and antioxidants are protective.


Acid-base assessment of post-parturient German Holstein dairy cows from jugular venous blood and urine: A comparison of the strong ion approach and traditional blood gas analysis.
Evaluating acid-base status is important for monitoring dairy herd health. In a field study, we aimed to compare the acid-base status measured by net acid-base excretion (NABE) in urine with results of venous blood analysis in clinically healthy, but possibly metabolically burdened cows in their transition period. For this, we sampled blood from the jugular vein and urine from 145 German Holstein cows within 1 to 76 days post-partum. In blood, the metabolic parameters non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHB), as well as numerous parameters of the acid-base status were measured. The traditional approach, based on bicarbonate concentration, base excess (BE) and anion gap (AG), was compared to the strong ion approach variables, e.g. acid total (Atot), measured strong ion difference (SIDm), strong ion gap (SIG), and unmeasured anions (XA), respectively. Results of both approaches were set against the outcome of urine analysis, i.e. the NABE, base-acid ratio and pH of urine, in a cluster analysis, which provided 7 moderately stable clusters. Evaluating and interpreting these 7 clusters offered novel insights into the pathophysiology of the acid-base equilibrium in fresh post-partum dairy cows. Especially in case of subclinical acid-base disorders, the parameters of the strong ion difference theory, particularly SIDm, Atot and SIG or XA, provided more in-depth information about acid-base status than the traditional parameters BE, bicarbonate or AG in blood. The acid-base status of fresh cows with protein aberrations in blood could be differentiated in a much better way using the strong ion approach than by traditional blood gas analysis or by the measurement of urinary excretion. Therefore, the strong ion approach seems to be a suitable supplement for monitoring acid-base balance in dairy cattle.


Development of HIV vectors for anti-HIV gene therapy.
Current gene therapy protocols for HIV infection use transfection or murine retrovirus mediated transfer of antiviral genes into CD4+ T cells or CD34+ progenitor cells ex vivo, followed by infusion of the gene altered cells into autologous or syngeneic/allogeneic recipients. While these studies are essential for safety and feasibility testing, several limitations remain: long-term reconstitution of the immune system is not effected for lack of access to the macrophage reservoir or the pluripotent stem cell population, which is usually quiescent, and ex vivo manipulation of the target cells will be too expensive and impractical for global application. In these regards, the lentivirus-specific biologic properties of the HIVs, which underlie their pathogenetic mechanisms, are also advantageous as vectors for gene therapy. The ability of HIV to specifically target CD4+ cells, as well as non-cycling cells, makes it a promising candidate for in vivo gene transfer vector on one hand, and for transduction of non-cycling stem cells on the other. Here we report the use of replication-defective vectors and stable vector packaging cell lines derived from both HIV-1 and HIV-2. Both HIV envelopes and vesicular stomatitis virus glycoprotein G were effective in mediating high-titer gene transfer, and an HIV-2 vector could be cross-packaged by HIV-1. Both HIV-1 and HIV-2 vectors were able to transduce primary human macrophages, a property not shared by murine retroviruses. Vesicular stomatitis virus glycoprotein G-pseudotyped HIV vectors have the potential to mediate gene transfer into non-cycling hematopoietic stem cells. If so, HIV or other lentivirus-based vectors will have applications beyond HIV infection.


HMMER web server: interactive sequence similarity searching.
HMMER is a software suite for protein sequence similarity searches using probabilistic methods. Previously, HMMER has mainly been available only as a computationally intensive UNIX command-line tool, restricting its use. Recent advances in the software, HMMER3, have resulted in a 100-fold speed gain relative to previous versions. It is now feasible to make efficient profile hidden Markov model (profile HMM) searches via the web. A HMMER web server (http://hmmer.janelia.org) has been designed and implemented such that most protein database searches return within a few seconds. Methods are available for searching either a single protein sequence, multiple protein sequence alignment or profile HMM against a target sequence database, and for searching a protein sequence against Pfam. The web server is designed to cater to a range of different user expertise and accepts batch uploading of multiple queries at once. All search methods are also available as RESTful web services, thereby allowing them to be readily integrated as remotely executed tasks in locally scripted workflows. We have focused on minimizing search times and the ability to rapidly display tabular results, regardless of the number of matches found, developing graphical summaries of the search results to provide quick, intuitive appraisement of them.


Stability of AML1 (core) site enhancer mutations in T lymphomas induced by attenuated SL3-3 murine leukemia virus mutants.
Murine retrovirus SL3-3 is highly T lymphomagenic. Its pathogenic properties are determined by the transcriptional enhancer of the U3 repeat region which shows preferential activity in T cells. Within the U3 repeats, the major determinant of T-cell specificity has been mapped to binding sites for the AML1 transcription factor family (also known as the core binding factor [CBF], polyomavirus enhancer binding protein 2 [PEBP2], and SL3-3 enhancer factor 1 [SEF-1]). SL3-3 viruses with AML1 site mutations have lost a major determinant of T-cell-specific enhancer function but have been found to retain a lymphomagenic potential, although disease induction is slower than for the SL3-3 wild type. To compare the specificities and mechanisms of disease induction of wild-type and mutant viruses, we have examined lymphomas induced by mutant viruses harboring transversions of three consecutive base pairs critical to AML1 site function (B. Hallberg, J. Schmidt, A. Luz, F. S. Pedersen, and T. Grundström. J. Virol. 65:4177-4181, 1991). Our results show that the mutated AML1 sites are genetically stable during lymphomagenesis and that ecotropic provirus numbers in DNA of tumors induced by wild-type and mutant viruses fall within the same range. Moreover, proviruses were found to be integrated at the c-myc locus in similar proportions of wild-type and mutant SL3-3-induced tumors, and the mutated AML1 sites of proviruses at c-myc are unaltered. In some cases, however, including one c-myc-integrated provirus, a single-base pair change was detected in a second, weaker AML1 binding site. By DNA rearrangement analysis of the T-cell receptor beta-locus, tumors induced by the AML1 site mutants are found to be of the T-cell type. Thus, although the AML1 site mutants have weakened T-cell-specific enhancers they are T-lymphomagenic, and wild-type- and mutant-virus-induced tumor DNAs are similar with respect to the number of overall ecotropic and c-myc-integrated clonal proviruses. The SL3-3 wild-type and AML1 site mutant viruses may therefore induce disease by similar mechanisms.


A Two-Moment Inequality with Applications to Rényi Entropy and Mutual Information.
This paper explores some applications of a two-moment inequality for the integral of the rth power of a function, where 0<r<1. The first contribution is an upper bound on the Rényi entropy of a random vector in terms of the two different moments. When one of the moments is the zeroth moment, these bounds recover previous results based on maximum entropy distributions under a single moment constraint. More generally, evaluation of the bound with two carefully chosen nonzero moments can lead to significant improvements with a modest increase in complexity. The second contribution is a method for upper bounding mutual information in terms of certain integrals with respect to the variance of the conditional density. The bounds have a number of useful properties arising from the connection with variance decompositions.


Nemaline rod disease, with reference to the routine use of histochemical methods in autopsy investigations.
A 50-year-old man had suffered from an undiagnosed form of muscle disease for most of his life. Death was due to cardiac failure. Numerous samples of muscle, obtained at autopsy, were subjected to the usual histochemical techniques used in investigating neuromuscular disease. Frozen sections revealed the abnormal accumulations of Z-band protein that characterize nemaline rod disease. That this disorder would have escaped diagnosis had only routine paraffin-embedded sections been examined is emphasized.


Simulation of keratoconus observation in photorefraction.
In the recent years, keratoconus (KC) has increasingly gained attention due to its treatment options and to the popularity of keratorefractive surgery. This paper investigates the potential of identification of KC using photorefraction (PR), an optical technique that is similar to objective retinoscopy and is commonly used for large-scale ocular screening. Using personalized eye models of both KC and pre-LASIK patients, computer simulations were performed to achieve visualization of this ophthalmic measurement. The simulations are validated by comparing results to two sets of experimental measurements. These PR images show distinguishable differences between KC eyes and eyes that are either normal or ametropic. The simulation technique with personalized modeling can be extended to other ophthalmic instrument developments. It makes possible investigation with the least number of real human subjects. The application is also of great interest in medical training.


[Lipoic acid as a means of metabolic therapy of open-angle glaucoma].
A total of 45 patients (90 eyes) with stages I and II open-angle glaucoma (OAG) were examined, 26 of these were administered lipoic acid in a daily dose of 0.075 g for 2 months and 19 were given 0.15 g daily for 1 month. Control group consisted of 31 patients with OAG who were administered only local hypotensive therapy. Vision acuity and visual field were checked up, tonometry, tonography, and campimetry carried out, and levels of nonprotein SH-groups and activity of gamma-glutamyl transpeptidase measured in the lacrimal fluid. Improvement of the biochemical parameters, visual function, and of the coefficient of efficacy of liquid discharge was more expressed in the patients administered lipoic acid in a daily dose of 0.15 g. Color campimetry results indicate improved sensitivity of the visual analyzer under the effect of treatment. Improvement was attained in approximately 45-47.5% of examined eyes, and was more often seen in patients with stage II OAG: in 57-58% eyes. The effect of lipoic acid may be explained by its antioxidant properties and direct influence on ocular tissue metabolism.


Comparison of novel oral anticoagulants versus vitamin K antagonists in patients with chronic kidney disease.
Novel oral anticoagulants (NOACs) including apixaban, dabigatran and rivaroxaban have been approved by international regulatory agencies to prevent venous thromboembolism as well as treat atrial fibrillation and venous thromboembolism in individuals with chronic kidney disease (CKD). However, alterations in their metabolism in the setting of CKD may impact their efficacy and lead to an increased risk of bleeding. This review summarizes the current literature on the efficacy and safety of these agents in individuals with moderate CKD. In clinical trials, the use of the NOACs in patients with moderate CKD has demonstrated efficacy and safety similar to those seen with vitamin K antagonists. However, no universal reversal agent for the anticoagulant effect of the NOACs exists in the setting of bleeding. Limited data have demonstrated that hemodialysis has been effectively used to aid in reversing the effects of dabigatran, and the use of prothrombin complex concentrate has also been used for serious and major adverse bleeding events with some success. As the use of the NOACs in patients with CKD increases, it will be important to monitor their safety, and clinicians who prescribe them should carefully monitor kidney function and recognize the potential for adverse effects.


Leech removal is not the primary driver of basking behavior in a freshwater turtle.
Leaving the water to bask (usually in the sun) is a common behavior for many freshwater turtles, with some species also engaging in "nocturnal basking." Ectoparasite removal is an obvious hypothesis to explain nocturnal basking and has also been proposed as a key driver of diurnal basking. However, the efficacy of basking, day or night, to remove leeches has not been experimentally tested. Therefore, we examined the number of leeches that were removed from Krefft's river turtles (Emydura macquarii krefftii) after experimentally making turtles bask at a range of times of day, durations, and temperatures. Turtles had high initial leech loads, with a mean of 32.1 leeches per turtle. Diurnal basking under a heat lamp for 3 hr at ~28°C significantly reduced numbers of leeches relative to controls. In diurnal trials, 90.9% of turtles lost leeches (mean loss of 7.1 leeches per turtle), whereas basking for 30 min under the same conditions was not effective (no turtles lost leeches, and all turtles were still visibly wet). Similarly, "nocturnal basking" at ~23°C for 3 hr was not effective at removing leeches. Only 18% of turtles lost leeches (one turtle lost one leech and another lost four leeches). Diurnal basking outdoors under direct sunlight for 20 min (mean temp = 34.5°C) resulted in a small reduction in leeches, with 50% of turtles losing leeches and an average loss of 0.7 leeches per turtle. These results indicate basking can remove leeches if temperatures are high or basking durations are long. However, it was only effective at unusually long basking durations in this system. Our data showed even the 20-min period was longer than 70.1% of natural diurnal basking events, many of which took place at cooler temperatures. Therefore, leech removal does not appear to be the purpose of the majority of basking events.


Identification of individual tetra- and pentacyclic naphthenic acids in oil sands process water by comprehensive two-dimensional gas chromatography/mass spectrometry.
The oils sands industry of Canada produces large volumes of process water (OSPW) which is stored in large lagoons. The OSPW contains complex mixtures of somewhat toxic, water-soluble, acid-extractable organic matter sometimes called 'naphthenic acids' (NA). Concerns have been raised over the possible environmental impacts of leakage of OSPW and a need has therefore arisen for better characterisation of the NA. Recently, we reported the first identification of numerous individual tricyclic NA in OSPW by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry (GCxGC/ToF-MS) of the methyl esters. The acids were diamondoid adamantane acids, resulting, it was proposed, from biotransformation of the corresponding alkyladamantane hydrocarbons, which is a known process. Biotransformation of higher alkylated diamondoid hydrocarbons was, until now, unknown but here we describe the identification of numerous pentacyclic NA as diamantane and alkyldiamantane acids, using the same methods. Further, we suggest tentative structures for some of the tetracyclic acids formed, we propose, by ring-opening of alkyldiamantanes. We suggest that this is further evidence that some of the acid-extractable organic matter in the OSPW originates from extensive biodegradation of the oil, whether in-reservoir or environmental, although other oxidative routes (e.g. processing) may also be possible. The results may be important for helping to better focus reclamation and remediation strategies for NA and for facilitating the identification of the sources of NA in contaminated environmental samples.


Beyond the hospital doors: Improving long-term outcomes for elderly trauma patients.
Elderly trauma patients (TPs) are the fastest growing trauma population, increasing the need for postacute care rehabilitation. For TP, discharge to skilled nursing facilities (SNFs) has been associated with higher 1-year mortality compared with discharge to inpatient rehabilitation facilities (IRFs) or home. The availability of IRF beds has been decreasing, but the proportion occupied by non-TPs, specifically stroke patients (SPs), has increased. We wanted to better characterize trends in trauma discharges and compare them with a population that is equally dependent on postdischarge rehabilitation. We hypothesized that discharge to SNF is rapidly increasing, while discharge to IRF is declining for trauma, but not for SPs. This is retrospective cohort study of adult trauma and SPs discharged from 2003 to 2009. The National Trauma Data Bank and National Inpatient Sample were used to study TPs and SPs, respectively. Falls became the leading cause of injury, and the proportion of older TPs increased from 23% to 30%. Older TPs discharged to SNF increased from 30.7% in 2003 to 40.8% in 2009 (p < 0.001). TPs were 34% (adjusted relative risk [RR], 1.34; 95% confidence interval [CI], 1.15-1.57) more likely to be discharged to an SNF and 36% (adjusted RR, 0.64; 95% CI, 0.48-0.85) less likely to be discharged to an IRF. From 2003 to 2009, SPs were 78% more likely to be discharged to an IRF (adjusted RR, 1.78; 95% CI, 1.74-1.82). The largest absolute increase in SP discharges to IRFs occurred the year following implementation of the stroke center certification program. For TPs, there was a significant increase in SNF discharges and a decrease in IRF discharges. During the same period, after implementation of stroke center certification, SPs were more likely to be discharged to an IRF. Future research should focus on evaluating which postacute care setting is most effective in providing rehabilitation for TPs and adjusting our discharge efforts to improve long-term outcomes. Prognostic and epidemiologic study, level III.


Spatiotemporal segregation in visual search: evidence from parietal lesions.
The mechanisms underlying segmentation and selection of visual stimuli over time were investigated in patients with posterior parietal damage. In a modified visual search task, a preview of old objects preceded search of a new set for a target while the old items remained. In Experiment 1, control participants ignored old and prioritized new items, but patients had severe difficulties finding the target (especially on the contralesional side). In Experiment 2, simplified displays yielded analogous results, ruling out search ease as a crucial factor in poor preview search. In Experiment 3, outlines around distractor groups (to aid segmentation) improved conjunction but not preview search, suggesting a specific deficit in spatiotemporal segmentation. Experiment 4 ruled out spatial disengagement problems as a factor. The data emphasize the role of spatiotemporal segmentation cues in preview search and the parietal lobe in the role of these cues to prioritize search of new stimuli.


Vertical Facial Skeletal Asymmetry and Occlusal Cant Relationship in Patients With Unilateral Cleft Lip and Palate.
To evaluate the maxillary and mandibular vertical skeletal asymmetries, and the correlation between these asymmetries and occlusal cant in patients with unilateral cleft lip-palate (UCLP). Anteroposterior radiographs of 25 patients with UCLP (UCLP group, mean age: 20.98 ± 4.88 years) and 25 subjects without cleft (control group, mean age: 19 ± 2.86 years) were included. Independent samples t test, Mann-Whitney U test and Pearson correlation analysis were performed based on linear and angular measurements. Lower facial horizontal asymmetry did not show statistically significant difference between the UCLP and control groups. However, vertical asymmetry of (a) the lateral cranial base (P = .014), (b) the nasomaxillary region (P < .001), (c) the maxillary dentoalveolus (P = .001), and (d) the lower face (P = .038) were all found to be significantly greater in UCLP group. The occlusal cant angle was also significantly greater in patients with UCLP compared to the controls (P = .016). While the occlusal cant angle was found to be correlated with the vertical asymmetry of the occlusal cant (r = 0.931, P < .001), maxillary cant angle was found to be correlated with the vertical asymmetry of the maxillary dentoalveolus (r = 0.655, P < .001). There was no correlation between the occlusal cant and the vertical and horizontal skeletal asymmetries. Vertical asymmetries of the lower face and the medial cranial base were negatively correlated with the horizontal lower facial asymmetry.


Differential effects of immunosuppressants on lymphocyte function.
In vitro and in vivo parameters of T lymphocyte function were evaluated in guinea pigs following treatment with the "cycle-active" drugs, 6-mercaptopurine (6MP) and methotrexate, and the "non-cycle-active" alkylating agent, cyclophosphamide. Commencing at the time of sensitization to tuberculin protein, animals were treated with an 8 day course of one of the cytotoxic drugs. This regimen either reduced or abolished the cutaneous response to PPD. The two cycle-active drugs inhibited the in vitro lymphoproliferative response to PPD and suppressed the elaboration of migration inhibition factor (MIF) by lymph node cells. However, these agents did not reduce blood lymphocytes, deplete the cellularity of the thymic dependent areas of peripheral tissues, or alter the in vitro response of lymph node cells to the nonspecific mitogen PHA. In contrast, treatment with cyclophosphamide was associated with a reduction in peripheral blood and tissue lymphocytes and impaired responses to PHA by residual lymph node cells. In vitro proliferative responses to PPD were inhibited but the capacity of lymph node cells to elaborate MIF was not suppressed. In addition to their effects on antigen-reactive lymphocytes, all three drugs significantly reduced the number of macrophages in induced peritoneal exudates. With respect to immunosuppressive activities, results of these investigations suggest that the noncycle-active agents affect both intermitotic and dividing T lymphocytes without impairing certain intermitotic functions of residual cells. The cycle-active drugs have a more restricted toxicity limited to those T lymphocytes which have been stimulated to undergo active DNA synthesis by antigenic challenge.


[The gastric mucosal barrier and the abomasal ulcer in veal calves].
Besides a review of the literature on the normal structure and function of the abomasum and abomasal ulcers in veal calves the present report contains the findings resulting from investigations on the pathogenesis of antral lesions. Histologically, the antral lesions consisted of focal acute superficial coagulation necrosis and erosions of the mucosa (18 cases), and subacute ulcers (5 cases). Histochemically, the pyloric mucosae of the normal abomasums were characterized by the presence of varying amounts of epithelial mucins. A similar variation in the distribution of mucins was present in the antral mucosae of the abnormal abomasums outside the areas showing antral lesions. The epithelium bordering on mucosal lesions was frequently completely depleted of mucins, whereas peripherally the mucin content of the epithelium showed gradual increase. The histological and histochemical findings suggest an adequate mucous barrier, which may be an important factor in the pathogenesis of abomasal ulcers in veal calves.


Induction of antibodies to hyaluronic acid by immunization of rabbits with encapsulated streptococci.
The immunogenicity of hyaluronic acid was investigated. Rabbits were immunized with encapsulated group A and C streptococci. Intact long-chain hyaluronate was conjugated to BSA for use as antigen in an ELISA. Antibodies to the hyaluronate-BSA conjugate were detected in peak immune sera. The specificity of the antibodies for both mammalian and streptococcal hyaluronate was shown by inhibition studies. To further confirm the presence of antihyaluronate antibodies, hyaluronidase-digested streptococcal hyaluronate was conjugated to biotin and used as an antigen in the ELISA. A clear immunization effect was shown for each rabbit by the study of preimmune and postimmunization bleedings. Titers for each rabbit increased by greater than 32 - 256 - fold. Inhibition studies using hyaluronidase-digested hyaluronate and periodate-treated hyaluronate showed that the immunodominant site of antibody reactivity was a terminal glucuronic acid residue. Further studies showed that the carboxyl group of the terminal glucuronide was the major immunoreactive site. Both mammalian and streptococcal hyaluronate inhibited the immune rabbit sera reaction to streptococcal hyaluronate, demonstrating crossreactivity of these molecules. Thus, hyaluronate was shown to be immunogenic in rabbits.


Displacement and tuberculosis: recognition in nursing care.
Through a qualitative of study of public health nurses and their clients in Toronto, Canada, displacement is described as a central feature in illness and care. Specifically, how client experiences are anchored in circumstances of international migration and contagiousness is described, along with how nurses take up understandings of displacement as part of their daily work through diminishing the social displacement of immigration, connecting here and there, and minimizing the displacement of contagiousness. The research illustrates how responding to the implacing and displacing elements of the illness experience and broader lifecourse can be an important part of professional practice. Moreover it highlights how place sensitive geographical inquiry is insightful with respect to understanding the interplay between illness, wellbeing, and health care work.


Optical implementation of iterative fractional Fourier transform algorithm.
An optical implementation of iterative fractional Fourier transform algorithm is proposed and demonstrated. In the proposed implementation, the phase-shifting digital holography technique and the phase-type spatial light modulator are adopted for the measurement and the modulation of complex optical fields, respectively. With the devised iterative fractional Fourier transform system, we demonstrate two-dimensional intensity distribution synthesis in the fractional Fourier domain and three-dimensional intensity distribution synthesis simultaneously forming desired intensity distributions at several multi-focal planes.


[Anesthesia in heart surgery].
Today, the performance of anaesthesia in cardiac surgery is on a high quality level. It requires special knowledge of the various operative techniques, the different methods of extracorporeal circulation and haemodynamic changes of the underlying cardiological disease, as well as extensive experience in cardiac and vasoactive drugs. Heart patients are now being operated on in many departments of cardiac surgery. Over and above this patients with more or less severe cardiac diseases are being looked after in every Department of Anaesthesiology, thus, every anaesthesiologist should acquaint himself with these specific problems during his specialist training.


Disease distribution and medical resources during the Beijing 2008 Olympic and Paralympic Games.
Appropriate planning and staffing for medical services at large-scale athletic events is essential to provide for a safe and successful competition. There are few well-documented accounts describing the demand for such services. The present study provided the data from the Beijing 2008 Olympics and Paralympics, with a view to provide the guidance for planning future events. A total of 22 029 and 8046 patients, who received medical care from a physician at an Olympic or Paralympic medical station, were included. The patient proportion among different personnel, various disease proportions at different kinds of venues, and the disease spectrum at specified venues at the Olympics and Paralympics were analyzed. At both games, the patient proportion varied by accreditation status. The staff accounted for the largest number of visits at the Olympics (44.83%) and Paralympics (36.95%), with respiratory diseases the most common. Various disease spectrums were discovered at the different kinds of venues. Surgical diseases were the most frequently listed reason for visits, both at competition and non-competition venues, especially during the Paralympics. The sport-related injuries accounted for a majority of the surgical cases during both games. At training venues, ear nose and throat diseases accounted for the greatest number of visits during both games. During both games, people contracted different diseases at different venues. Adequate surgeons should be designated to offer assistance mostly in trauma situations. Appropriate numbers of physicians in respiratory diseases and otorhinolaryngology is of great importance.


Astaxanthin protects against pirarubicin-induced H9c2 cardiomyocytes by adjusting microRNA-494-3p-mediated MDM4/p53 signalling pathway.
Pirarubicin (THP) is an antitumour drug widely used in clinical practice, but its cardiotoxicity limits its application. THP cardiotoxicity must be treated as soon as possible. There is an urgent need to find drugs that alleviate THP cardiotoxicity. The purpose of this study was to investigate the effects and mechanisms of Astaxanthin (AST) on THP-induced cardiomyocytes. Rat cardiomyocytes H9c2 were induced with THP. The effects of AST on THP-induced H9c2 and its mechanism were investigated by CCK8, reactive oxygen species assay, tunnel assay, flow cytometry, RT-qPCR, and Western blot. AST increased cell viability, inhibited apoptosis and accelerated cell cycle progression, reduced oxidative damage and inflammatory response in THP-induced H9c2; down-regulated miR-494-3p expression, promoted MDM4 expression, inhibited p53 activation, and suppressed apoptosis-related protein expression. Overexpression of MiR-494-3p reversed the above effects of AST. AST can inhibit H9c2 apoptosis induced by THP and attenuate H9c2 damage by THP, which may be achieved by downregulating miR-494-3p, upregulating MDM4, and inhibiting p53.


All-Cause and Cause-Specific Mortality in Psoriatic Arthritis and Ankylosing Spondylitis: A Systematic Review and Meta-Analysis.
Psoriatic arthritis (PsA) and ankylosing spondylitis (AS) are chronic inflammatory diseases associated with a higher risk of cardiometabolic comorbidities compared to the general population. Individual studies examining mortality in these patients have produced conflicting results. The present study was undertaken to perform a systematic review and meta-analysis to analyze the all-cause and cause-specific mortality in PsA and AS from the available literature. A comprehensive database search was performed for studies reporting all-cause or cause-specific mortality in patients with PsA and AS compared with the general population. Pooled relative risks (RRs) were calculated using a random-effects model. We included 19 studies (11 of PsA, 7 of AS, 1 of both). In PsA studies, there was no increased mortality compared to the general population (RR 1.12 [95% confidence interval (95% CI) 0.96-1.30]; n = 10 studies). We found a higher all-cause mortality in female (RR 1.19 [95% CI 1.04-1.36]) but not in male (RR 1.02 [95% CI 0.66-1.59]) PsA patients. Cardiovascular-, respiratory-, and infection-specific mortality risks were significantly higher for PsA patients (RR 1.21 [95% CI 1.06-1.38], RR 3.37 [95% CI 1.30-8.72], and RR 2.43 [95% CI 1.01-5.84], respectively), but not cancer-related mortality (RR 1.01 [95% CI 0.91-1.11]). In AS, we found a higher risk of death from all causes (RR 1.64 [95% CI 1.49-1.80]; n = 6 studies) and cardiovascular causes (RR 1.35 [95% CI 1.01-1.81]; n = 3 studies) compared to the general population. All-cause mortality was high in both male (RR 1.56 [95% CI 1.43-1.71]) and female (RR 1.85 [95% CI 1.56-2.18]) AS patients. The included AS studies did not report mortality data for non-cardiovascular causes. This systematic review and meta-analysis showed a significantly increased risk of overall mortality in AS but not PsA. Cardiovascular-specific mortality was higher for both PsA and AS, which emphasizes the importance of early screening and management of cardiovascular risk factors.


Expressional divergences of two desaturase genes determine the opposite ratios of two sex pheromone components in Helicoverpa armigera and Helicoverpa assulta.
The sympatric closely related species Helicoverpa armigera and Helicoverpa assulta use 97:3 and 7:93 of (Z)-11-hexadecenal and (Z)-9-hexadecenal, respectively, as their sex pheromone to find/locate correct sex mates. Moreover, (Z)-11-hexadecenyl alcohol and (Z)-9-hexadecenyl alcohol are more abundant in the pheromone gland of H. assulta than in that of H. armigera. To clarify the molecular basis of these differences, we sequenced the pheromone gland transcriptomes of the two species and compared the expression patterns of the candidate enzyme genes involved in the pheromone biosynthetic pathways by FPKM values and quantitative RT-PCR analysis. We found that the desaturase gene LPAQ expressed about 70 times higher in H. armigera than in H. assulta, whereas another desaturase gene NPVE expressed about 60 times higher in H. assulta than in H. armigera. We also observed significantly higher expression of the fatty acyl reductase (FAR) gene FAR1 and the aldehyde reductase (AR) gene AR3 in H. assulta than in H. armigera. Examination of the pheromone glands of the backcross offspring of their hybrids to H. assulta showed a positive linear correlation between the expression level of LPAQ and the amount of Z11-16:Ald and between the expression level of NPVE and the amount of Z9-16:Ald in the pheromone glands. Taken together, these data demonstrate that the expressional divergences of LPAQ and NPVE determine the opposite sex pheromone component ratios in the two species and the divergent expression of FAR1 and AR3 may account for the greater accumulation of alcohols in the pheromone gland of H. assulta.


Evaluation of the viability of bone fragments.
We used laser Doppler flowmetry (LDF) to measure flux in cortical bone fragments as a method of determining their vascular status and viability. In an experimental tibial osteotomy measurements of flux were made from specific cortical sites both before and after osteotomy. Flux levels fell rapidly in non-vascularised fragments and remained significantly reduced throughout the experiment. By contrast, those in vascularised fragments were significantly reduced one and two hours after the osteotomy but then increased. From three hours after the osteotomy there was no significant difference in flux levels between the vascularised fragments and proximal bone stock. We conclude that measurement of bone flux by LDF may have a role in the objective evaluation of the viability of bone fragments, but that further studies are required to validate the technique before its adoption in the management of the injured patient.


The importance of plant diversity in maintaining the pollinator bee, Eulaema nigrita (Hymenoptera: Apidae) in sweet passion fruit fields.
The euglossine bee Eulaema nigrita plays an important role for the pollination of native and economically important plants, such as the sweet passion-fruit Passiflora alata. E. nigrita uniquely collects the nectar from the flowers of P. alata, nevertheless, it needs to visit other plants to collect pollen, nectar and other resources for its survival. There are two methods to identify the species of plants used by bees in their diet: by direct observation of the bees in the flowers, and through identification of pollen grains present in brood cells, feces, or in the bees' body. In order to identify the other plants that E. nigrita visits, we analyzed samples of pollen grains removed from the bee's body in the course of the flowering period of P. alata. Among our results, the flora visited by E. nigrita comprised 40 species from 32 genera and 19 families, some of them used as a pollen source or just nectar. In spite of being a polyletic species, E. nigrita exhibited preference for some plant species with poricidal anthers. P. alata which has high sugar concentration nectar was the main source of nectar for this bee in the studied area. Nonetheless, the pollinic analysis indicated that others nectariferous plant species are necessary to keep the populations of E. nigrita. Studies such as this one are important since they indicate supplementary pollen-nectar sources which must be used for the conservation of the populations of E. nigrita in crops neighbouring areas. In the absence of pollinators, growers are forced to pay for hand pollination, which increases production costs; keeping pollinators in cultivated areas is still more feasible to ensure sweet passion fruit production.


Multidimensional (co)evolutionary stability.
The complexity of biotic and abiotic environmental conditions is such that the fitness of individuals is likely to depend on multiple traits. Using a synthetic framework of phenotypic evolution that draws from adaptive dynamics and quantitative genetics approaches, we explore how the number of traits under selection influences convergence stability and evolutionary stability in models for coevolution in multidimensional phenotype spaces. Our results allow us to identify three different effects of trait dimensionality on stability. First are (i) a "combinatorial effect": without epistasis and genetic correlations, a higher number of trait dimensions offers more opportunities for equilibria to be unstable; and (ii) epistatic interactions, that is, fitness interactions between traits, which tend to destabilize evolutionary equilibria; this effect increases with the dimension of phenotype space. These first two effects influence both convergence stability and evolutionary stability, while (iii) genetic correlations (due, e.g., to pleiotropy or linkage disequilibrium) can affect only convergence stability. We illustrate the general prediction that increased dimensionality destabilizes evolutionary equilibria using examples drawn from well-studied classical models of frequency-dependent competition for resources, adaptation to a spatially heterogeneous environment, and antagonistic coevolution. In addition, our analyses show that increased dimensionality can favor diversification, for example, in the form of local adaptation, as well as evolutionary escape.


Revisiting physician impact analyses: predicted versus observed hospital resource use.
In order to determine the accuracy of predictions made using Physician Impact Analysis (PIA), a comparison was undertaken of predicted versus actual resource use for 10 new physician specialists at a southern Ontario community hospital. The predictions were done from 1987 to 1989 using methodology recommended by the Ontario Hospital Association (OHA) and available at that time. This included (1) Hospital Medical Records Institute data and (2) a hospital service department survey. A comparison was made between PIA predictions and actual physician resource utilization data gathered in 1991. There was little agreement between the two. The usefulness of PIA rests largely on its ability to generate accurate predictions about a new physician recruit's number of cases and hospital resource use. However, because this research was undertaken in only one community hospital, further evaluation of the PIA process is recommended.


Isolation of a Chinese hamster ovary cell mutant defective in intramitochondrial transport of phosphatidylserine.
A CHO-K1 cell mutant with a specific decrease in cellular phosphatidylethanolamine (PE) level was isolated as a variant resistant to Ro09-0198, a PE-directed antibiotic peptide. The mutant was defective in the phosphatidylserine (PS) decarboxylation pathway for PE formation, in which PS produced in the endoplasmic reticulum is transported to mitochondria and then decarboxylated by an inner mitochondrial membrane enzyme, PS decarboxylase. Neither PS formation nor PS decarboxylase activity was reduced in the mutant, implying that the mutant is defective in some step of PS transport. The transport processes of phospholipids between the outer and inner mitochondrial membrane were analyzed by use of isolated mitochondria and two fluorescence-labeled phospholipid analogs, 1-palmitoyl-2-[N-[6(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]caproyl]-PS (C6-NBD-PS) and C6-NBD-phosphatidylcholine (C6-NBD-PC). On incubation with the CHO-K1 mitochondria, C6-NBD-PS was readily decarboxylated to C6-NBD-PE, suggesting that the PS analog was partitioned into the outer leaflet of mitochondria and then translocated to the inner mitochondrial membrane. The rate of decarboxylation of C6-NBD-PS in the mutant mitochondria was reduced to approximately 40% of that in the CHO-K1 mitochondria. The quantity of phospholipid analogs translocated from the outer leaflet of mitochondria into inner mitochondrial membranes was further examined by selective extraction of the analogs from the outer leaflet of mitochondria. In the mutant mitochondria, the translocation of C6-NBD-PS was significantly reduced, whereas the translocation of C6-NBD-PC was not affected. These results indicate that the mutant is defective in PS transport between the outer and inner mitochondrial membrane and provide genetic evidence for the existence of a specific mechanism for intramitochondrial transport of PS.


[Poseidonol in the treatment of patients with hyperlipidemia].
An antilipidemic effect of poseidonol was tested in 123 patients with ischemic heart disease, stable angina of effort (II-III functional class), 40 patients with secondary pyelonephritis and 40 patients with chronic hepatosis-hepatitis associated with chronic alcoholism or diabetes mellitus. The patients had also hyperlipidemia. A positive effect of poseidonol was established: cholesterol content fell by 25.4, 20.6 and 18.7%, respectively; triglycerides level decreased by 63, 42 and 21.4%, respectively; B-lipoproteins fell by 55.5, 36.9 and 13.1%, respectively. High density lipoproteins rose by 40.2, 27 and 69.9%, respectively. No adverse effects either on the liver or kidney functions were observed.


Real-time near-infrared fluorescence imaging using cRGD-ZW800-1 for intraoperative visualization of multiple cancer types.
Incomplete resections and damage to critical structures increase morbidity and mortality of patients with cancer. Targeted intraoperative fluorescence imaging aids surgeons by providing real-time visualization of tumors and vital structures. This study evaluated the tumor-targeted zwitterionic near-infrared fluorescent peptide cRGD-ZW800-1 as tracer for intraoperative imaging of multiple cancer types. cRGD-ZW800-1 was validated in vitro on glioblastoma (U-87 MG) and colorectal (HT-29) cell lines. Subsequently, the tracer was tested in orthotopic mouse models with HT-29, breast (MCF-7), pancreatic (BxPC-3), and oral (OSC-19) tumors. Dose-ranging studies, including doses of 0.25, 1.0, 10, and 30 nmol, in xenograft tumor models suggest an optimal dose of 10 nmol, corresponding to a human equivalent dose of 63 μg/kg, and an optimal imaging window between 2 and 24 h post-injection. The mean half-life of cRGD-ZW800-1 in blood was 25 min. Biodistribution at 4 h showed the highest fluorescence signals in tumors and kidneys. In vitro and in vivo competition experiments showed significantly lower fluorescence signals when U-87 MG cells (minus 36%, p = 0.02) or HT-29 tumor bearing mice (TBR at 4 h 3.2 ± 0.5 vs 1.8 ± 0.4, p = 0.03) were simultaneously treated with unlabeled cRGD. cRGD-ZW800-1 visualized in vivo all colorectal, breast, pancreatic, and oral tumor xenografts in mice. Screening for off-target interactions, cRGD-ZW800-1 showed only inhibition of COX-2, likely due to binding of cRGD-ZW800-1 to integrin αVβ3. Due to its recognition of various integrins, which are expressed on malignant and neoangiogenic cells, it is expected that cRGD-ZW800-1 will provide a sensitive and generic tool to visualize cancer during surgery.


Corticosteroids inhibit IL-12 production in human monocytes and enhance their capacity to induce IL-4 synthesis in CD4+ lymphocytes.
We examined the effects of corticosteroids on IL-12 production by human monocytes and on cytokine synthesis in T cells. To distinguish the effects of corticosteroids on the APC used to activate the T cell from direct effects of corticosteroids on the T cell, experiments were performed by exposing the APC and not the T cell to corticosteroids. We found that corticosteroids significantly inhibited the production in monocytes of IL-12, a cytokine that is extremely potent in enhancing IFN-gamma and inhibiting IL-4 synthesis in T cells. We demonstrated that reduced production of IL-12 in corticosteroid-treated monocytes resulted in a decreased capacity of the monocytes to induce IFN-gamma and an increased ability to induce IL-4 in T cells. These results suggest that although corticosteroids may be beneficial for the treatment of asthma or allergic disease due to direct inhibitory effects of corticosteroids on cytokine synthesis in T cells, chronic corticosteroid therapy may indirectly exacerbate the long-term course of allergic disease. This deleterious effect of corticosteroids would result from a limitation in IL-12 production in tissue monocytes and macrophages, which would enhance production of Th2 cytokines (which augment allergic disease), and would reduce production of Th1 cytokines (which attenuate allergic disease) in T cells that subsequently infiltrate the tissues.


Reduced virulence of Yersinia enterocolitica by copper-induced injury.
A sublethal concentration of copper (0.75 mg/liter) caused substantial injury (87 to 95%) of Yersinia enterocolitica serotype O:8 cells in 72 h at 4 degrees C without producing extensive cell death. Copper-injured cells had a higher 50% lethal dose in mice (2,700 CFU) than uninjured cells (150 CFU). This reduced virulence correlated with more rapid clearance of the injured cells from the blood of mice after intravenous inoculation. A possible role of the liver in this process was shown by significant cell accumulation in mouse livers when copper-injured Y. enterocolitica cells were administered, compared with uninjured bacteria. In vitro studies with isolated mouse liver membranes showed higher titers of aggregation with copper-injured cells than control cells. The in vitro aggregation reaction and blood clearance activity in vivo were abolished by sugars that are known to interact with a hepatic lectin. Our data suggest that copper-induced injury reduces the virulence of Y. enterocolitica and that the liver may be involved in nonimmune rapid clearance of the injured cells, probably by interaction with a hepatic lectin(s).


The Vc2 Cyclic di-GMP-Dependent Riboswitch of Vibrio cholerae Regulates Expression of an Upstream Putative Small RNA by Controlling RNA Stability.
Cyclic di-GMP (c-di-GMP) is a bacterial second messenger molecule that is important in the biology of Vibrio cholerae, but the molecular mechanisms by which this molecule regulates downstream phenotypes have not been fully characterized. We have previously shown that the Vc2 c-di-GMP-binding riboswitch, encoded upstream of the gene tfoY, functions as an off switch in response to c-di-GMP. However, the mechanism by which c-di-GMP controls expression of tfoY has not been fully elucidated. During our studies of this mechanism, we determined that c-di-GMP binding to Vc2 also controls the abundance and stability of upstream noncoding RNAs with 3' ends located immediately downstream of the Vc2 riboswitch. Our results suggest these putative small RNAs (sRNAs) are not generated by transcriptional termination but rather by preventing degradation of the upstream untranslated RNA when c-di-GMP is bound to Vc2.IMPORTANCE Riboswitches are typically RNA elements located in the 5' untranslated region of mRNAs. They are highly structured and specifically recognize and respond to a given chemical cue to alter transcription termination or translation initiation. In this work, we report a novel mechanism of riboswitch-mediated gene regulation in Vibrio cholerae whereby a 3' riboswitch, named Vc2, controls the stability of upstream untranslated RNA upon binding to its cognate ligand, the second messenger cyclic di-GMP, leading to the accumulation of previously undescribed putative sRNAs. We further demonstrate that binding of the ligand to the riboswitch prevents RNA degradation. As binding of riboswitches to their ligands often produces compactly structured RNA, we hypothesize this mechanism of gene regulation is widespread.


Parents' Views on Play and the Goal of Early Childhood Education in Relation to Children's Home Activity and Executive Functions: A Cross-Cultural Investigation.
The present study investigated the cross-cultural variations in parents' views on the role of play in child development and the primary purpose of preschool education from Ethiopia and Hungary. It also examined the cross-cultural variations in preschoolers' executive functions (EF), the frequency of their engagement in home activities, and the role of these activities in the development of EF skills. Participants included 266 preschoolers (Ethiopia: 139 of which 44.6% boys; M age = 63.83 months; SD = 7.68 months; Hungary: 127 of which 48% boys; M age = 62.06 months; SD = 9.37 months) with their parents (Ethiopia: 45.32% male; Mage = 36.66 years; SD = 7.14 years; Hungary: 13.18% male; M age = 37.71 years; SD = 5.97 years). The independent samples t-test showed that Ethiopian parents view fostering academic skills for preschooler significantly more important than their Hungarian counterparts do. We also found that while Ethiopian parents hold the belief that academic and cognitive development is the major purpose of preschool education, Hungarian prioritize social-emotional development and entertainment. Additionally, preschoolers in Ethiopia were reported to engage in academic and arts and crafts activities after preschool significantly more frequently than their Hungarian counterparts. On the contrary, preschoolers in Hungary were found to engage in fine-motor activities, solitary play, sports and other physical activities significantly more frequently than their Ethiopian counterparts. No significant differences were found in EF skills between preschoolers from Ethiopia and Hungary. Results from hierarchical regression analyses showed that, after accounting for age and SES, preschoolers' frequency of pretend play and their parents' play support beliefs were found to be small to medium-sized predictors of inhibitory control skills in both samples. However, children's frequency of having breakfast at home was another significant predictor in the Ethiopian sample only. The frequency of participation in arts and crafts and other fine-motor activities were found to be important predictors of preschoolers' visual-spatial working memory skills in the Ethiopian and the Hungarian samples, respectively. We also found that, after controlling for SES, parental play support was an important factor associated with preschoolers' shifting skills only in the Hungarian sample. Based on the findings, we made important conclusions.


Absorption of 193- and 213-nm laser wavelengths in sodium chloride solution and balanced salt solution.
To determine absorption coefficients for sodium chloride solution (saline) and balanced salt solution at the 193- and 213-nm laser wavelengths. Absorption coefficients were obtained for each of the component species found in balanced salt solution. This was achieved by measuring laser pulse transmission through solutions of varying concentration. The experiments were repeated using the 193-nm excimer and 213-nm solid-state laser wavelengths. Results for each species were then used to obtain an overall absorption coefficient and penetration depth for balanced salt solution and 0.9% sodium chloride solution. Absorption coefficients in balanced salt solution for the 193- and 213-nm wavelengths were found to be 140 and 6.9 cm(-1), respectively. In 0.9% sodium chloride solution, the absorption coefficient was 81 cm(-1) at 193 nm and 0.05 cm(-1) at 213 nm. At 193 nm, absorption in balanced salt solution was dominated by sodium chloride. Sodium citrate emerged as the dominant species of absorption at 213 nm. For the species investigated, we found reduced absorption for the longer wavelength of 213 nm. While the difference in wavelength between 193 and 213 nm is within about 10%, the respective molar absorption coefficients varied by 1 to 4 orders of magnitude. This indicates that predictions for the wavelength-dependent changes of absorption coefficients of other solutions are unreliable. Fluid placed on the surface of the cornea during keratorefractive surgery has proved to be a barrier to ablation for the 193-nm wavelength. The increased penetration depth through sodium chloride solution and balanced salt solution for the longer 213-nm laser wavelength may mean that these solutions cannot be used as a masking agent for keratorefractive procedures performed with this wavelength.


An immunohistochemical study of cases of sporadic and inherited frontotemporal lobar degeneration using 3R- and 4R-specific tau monoclonal antibodies.
The pathological distinctions between the various clinical and pathological manifestations of frontotemporal lobar degeneration (FTLD) remain unclear. Using monoclonal antibodies specific for 3- and 4-repeat isoforms of the microtubule associated protein, tau (3R- and 4R-tau), we have performed an immunohistochemical study of the tau pathology present in 14 cases of sporadic forms of FTLD, 12 cases with Pick bodies and two cases without and in 27 cases of familial FTLD associated with 12 different mutations in the tau gene (MAPT), five cases with Pick bodies and 22 cases without. In all 12 cases of sporadic FTLD where Pick bodies were present, these contained only 3R-tau isoforms. Clinically, ten of these cases had frontotemporal dementia and two had progressive apraxia. Only 3R-tau isoforms were present in Pick bodies in those patients with familial FTLD associated with L266V, Q336R, E342V, K369I or G389R MAPT mutations. Patients with familial FTLD associated with exon 10 N279K, N296H or +16 splice site mutations showed tau pathology characterised by neuronal neurofibrillary tangles (NFT) and glial cell tangles that contained only 4R-tau isoforms, as did the NFT in P301L MAPT mutation. With the R406W mutation, NFT contained both 3R- and 4R-tau isoforms. We also observed two patients with sporadic FTLD, but without Pick bodies, in whom the tau pathology comprised only of 4R-tau isoforms. We have therefore shown by immunohistochemistry that different specific tau isoform compositions underlie the various kinds of tau pathology present in sporadic and familial FTLD. The use of such tau isoform specific antibodies may refine pathological criteria underpinning FTLD.


Seven-Coordinate Tb3+ Complexes with 90% Quantum Yields: High-Performance Examples of Combined Singlet- and Triplet-to-Tb3+ Energy-Transfer Pathways.
Seven-coordinate, pentagonal-bipyramidal (PBP) complexes [Ln(bbpen)Cl] and [Ln(bbppn)Cl], in which Ln = Tb3+ (products I and II), Eu3+ (III and IV), and Gd3+ (V and VI), with bbpen2- = N,N'-bis(2-oxidobenzyl)-N,N'-bis(pyridin-2-ylmethyl)ethylenediamine and bbppn2- = N,N'-bis(2-oxidobenzyl)-N,N'-bis(pyridin-2-ylmethyl)-1,2-propanediamine, were synthesized and characterized by single-crystal X-ray diffraction analysis, alternating-current magnetic susceptibility measurements, and photoluminescence (steady-state and time-resolved) spectroscopy. Under a static magnetic field of 0.1 T, the Tb3+ complexes I and II revealed single-ion-magnet behavior. Also, upon excitation at 320 nm at 300 K, I and II presented very high absolute emission quantum yields (0.90 ± 0.09 and 0.92 ± 0.09, respectively), while the corresponding Eu3+ complexes III and IV showed no photoluminescence. Detailed theoretical calculations on the intramolecular energy-transfer rates for the Tb3+ products indicated that both singlet and triplet ligand excited states contribute efficiently to the overall emission performance. The expressive quantum yields, QLnL, measured for I and II in the solid state and a dichloromethane solution depend on the excitation wavelength, being higher at 320 nm. Such a dependence was rationalized by computing the intersystem crossing rates (WISC) and singlet fluorescence lifetimes (τS) related to the population dynamics of the S1 and T1 levels. Thin films of product II showed high air stability and photostability upon continuous UV illumination, which allowed their use as downshifting layers in a green light-emitting device (LED). The prototypes presented a luminous efficacy comparable with those found in commercial LED coatings, without requiring encapsulation or dispersion of II in host matrixes. The results indicate that the PBP environment determined by the ethylenediamine (en)-based ligands investigated in this work favors the outstanding optical properties in Tb3+ complexes. This work presents a comprehensive structural, chemical, and spectroscopic characterization of two Tb3+ complexes of mixed-donor, en-based ligands, focusing on their outstanding optical properties. They constitute good molecular examples in which both triplet and singlet excited states provide energy to the Tb3+ ion and lead to high values of QLnL.


A new animal model for bone atrophic nonunion: fixation by external fixator.
A new small animal model of bone atrophic nonunion was established for investigating the process of bone regeneration by performing cauterization of the periosteum, removal of the local bone marrow, and stabilization with external fixation. The model allows the creation of an atrophic nonunion without the need for a critical size defect. Furthermore, it provides reproducible, well-defined mechanical conditions and minimized physical interference of the implant with the biological processes in the healing zone. Eighty adult Sprague-Dawley rats received an osteotomy of the left femur, stabilized with an external fixator. In half of the animals, the periosteum proximal and distal to the osteotomy was destroyed by cauterization and the adjacent bone marrow was removed (nonunion group). At 2 and 8 weeks after surgery, radiological, biomechanical, histological, and histomorphometrical analyses showed a typical physiological healing in the control group, while the nonunion group was characterized by resorption of the bone ends with some callus formation distant to the osteotomy. At both time points, the callus was composed of significantly less bone and significantly more connective tissue (p < 0.001). In addition, the torsional strength of the osteotomized femur was significantly less in the nonunion group than in the control group, which was comparable to that of the intact femur (p < 0.001). In conclusion, the present model allows the induction of an atrophic nonunion without the need of a critical size defect. It is reproducible, provides standardized biomechanical conditions, and allows minimized interaction of the implant with the healing zone.


Lessons learned from the rapid development of a statewide simulation model for predicting COVID-19's impact on healthcare resources and capacity.
The first case of COVID-19 was detected in North Carolina (NC) on March 3, 2020. By the end of April, the number of confirmed cases had soared to over 10,000. NC health systems faced intense strain to support surging intensive care unit admissions and avert hospital capacity and resource saturation. Forecasting techniques can be used to provide public health decision makers with reliable data needed to better prepare for and respond to public health crises. Hospitalization forecasts in particular play an important role in informing pandemic planning and resource allocation. These forecasts are only relevant, however, when they are accurate, made available quickly, and updated frequently. To support the pressing need for reliable COVID-19 data, RTI adapted a previously developed geospatially explicit healthcare facility network model to predict COVID-19's impact on healthcare resources and capacity in NC. The model adaptation was an iterative process requiring constant evolution to meet stakeholder needs and inform epidemic progression in NC. Here we describe key steps taken, challenges faced, and lessons learned from adapting and implementing our COVID-19 model and coordinating with university, state, and federal partners to combat the COVID-19 epidemic in NC.


Sjögren-Larsson syndrome. Deficient activity of the fatty aldehyde dehydrogenase component of fatty alcohol:NAD+ oxidoreductase in cultured fibroblasts.
Sjögren-Larsson syndrome (SLS) is an inherited disorder associated with impaired fatty alcohol oxidation due to deficient activity of fatty alcohol:NAD+ oxidoreductase (FAO). FAO is a complex enzyme which consists of two separate proteins that sequentially catalyze the oxidation of fatty alcohol to fatty aldehyde and fatty acid. To determine which enzymatic component of FAO was deficient in SLS, we assayed fatty aldehyde dehydrogenase (FALDH) and fatty alcohol dehydrogenase in cultured fibroblasts from seven unrelated SLS patients. All SLS cells were selectively deficient in the FALDH component of FAO, and had normal activity of fatty alcohol dehydrogenase. The extent of FALDH deficiency in SLS cells depended on the aliphatic aldehyde used as substrate, ranging from 62% of mean normal activity using propionaldehyde as substrate to 8% of mean normal activity with octadecanal. FALDH activity in obligate SLS heterozygotes was partially decreased to 49 +/- 7% of mean normal activity using octadecanal as substrate. Differential centrifugation studies in fibroblasts indicated that this FALDH enzyme was largely particulate; soluble FALDH activity was normal in SLS cells. Intact SLS fibroblasts oxidized octadecanol to fatty acid at less than 10% of the normal rate, but oxidized free octadecanal normally, suggesting that the FALDH affected in SLS is chiefly involved in the oxidation of fatty alcohol to fatty acid. These results show that the primary enzymatic defect in SLS is the FALDH component of the FAO complex, which leads to deficient oxidation of fatty aldehyde derived from fatty alcohol.


The contribution of gluconeogenesis to glycogen repletion during glucose infusion in endotoxemia.
Glycogen repletion rates in liver and skeletal muscle were quantitated, and the contribution of gluconeogenesis to hepatic glycogen repletion and glucose output were determined during glucose infusion in hemodynamically stable endotoxemic animals. Four hours after the injection of endotoxin (or saline), rats were infused with 235 mumol/min/kg of glucose (or saline) containing [6-3H]-glucose for up to four additional hours. Glucose infusion increased the plasma glucose concentration, which plateaued between 14 to 17 mmol/L, in the control rats. The glucose concentration of the endotoxin group receiving glucose was consistently greater than in control rats and failed to reach a plateau. The rate of muscle glycogen synthesis was not altered after endotoxin, but hepatic glycogen repletion was decreased by 55%. The percentage of glycogen repletion derived directly from blood glucose in liver and muscle was similar in control and endotoxin-treated rats receiving glucose. Therefore, the direct incorporation of glucose into glycogen appeared to predominate over the contribution by gluconeogenesis. However, gluconeogenesis continued during the glucose infusion in both control and endotoxemic animals. The calculated rate of total gluconeogenesis was 160% higher in the endotoxin glucose-infused rats compared to the control animals receiving glucose. This increase was due primarily to the attenuated suppression of hepatic glucose output in the endotoxin group (52 +/- 4%) compared to controls (84 +/- 3%). Thus, gluconeogenically derived glucose-6-phosphate appears to be diverted from hepatic glycogen storage to glucose output in endotoxin-treated rats during glucose infusion.


Process inhomogeneity leads to rapid side product turnover in cultivation of Corynebacterium glutamicum.
Corynebacterium glutamicum has large scale industrial applications in the production of amino acids and the potential to serve as a platform organism for new products. This means the demand for industrial process development is likely to increase. However, large scale cultivation conditions differ from laboratory bioreactors, mostly due to the formation of concentration gradients at the industrial scale. This leads to an oscillating supply of oxygen and nutrients for microorganisms with uncertain impact on metabolism. Scale-down bioreactors can be applied to study robustness and physiological reactions to oscillating conditions at a laboratory scale. In this study, C. glutamicum ATCC13032 was cultivated by glucose limited fed-batch cultivation in a two-compartment bioreactor consisting of an aerobic stirred tank and a connected non-aerated plug flow reactor with optional feeding. Continuous flow through both compartments generated oscillating profiles with estimated residence times of 45 and 87 seconds in the non-aerated plug flow compartment. Oscillation of oxygen supply conditions at substrate excess and oscillation of both substrate and dissolved oxygen concentration were compared to homogeneous reference cultivations. The dynamic metabolic response of cells within the anaerobic plug flow compartment was monitored throughout the processes, detecting high turnover of substrate into metabolic side products and acidification within oxygen depleted zones. It was shown that anaerobic secretion of lactate into the extracellular culture broth, with subsequent reabsorption in the aerobic glucose-limited environment, leads to mixed-substrate growth in fed-batch processes. Apart from this, the oscillations had only a minor impact on growth and intracellular metabolite characteristics. Carbon metabolism of C. glutamicum changes at oscillating oxygen supply conditions, leading to a futile cycle over extracellular side products and back into oxidative pathways. This phenomenon facilitates a dynamic and flexible shift of oxygen uptake at inhomogeneous process conditions. There is no loss of process characteristics at oscillation times in the minute range, which emphasizes the robustness of C. glutamicum in comparison to other industrial microorganisms. Therefore, the metabolic phenotype of C. glutamicum seems to be particularly well-suited for cultivation at inhomogeneous process conditions for large-scale fed-batch application, which is in good accordance with the respective industrial experiences.


Low-temperature sterilization and new technologies.
The new low-temperature sterilization technologies are presented as an alternative to the use of ethylene oxide. This review was performed in order to identify evidences of the antimicrobial activity, toxicity, adverse events and the applicability of these technologies. The research was carried through the electronic databases MEDLINE and LILACS up to 2005. The authors analyzed 10 articles in this survey. The studies about the efficacy of these sterilization methods constitute experimental and comparative research that showed the influence of the extension and diameter of the lumen, besides the presence of crystal salts. Thus, choosing the correct equipment is essential, as well as the assurance of the cleansing of the devices, which interfere with the effectiveness of the low-temperature sterilization. These technologies present limitations regarding the sterilization of graft bone and affect the materials properties.


Massachusetts health reform and disparities in joint replacement use: difference in differences study.
To estimate the impact of the insurance expansion in 2006 on use of knee and hip replacement procedures by race/ethnicity, area income, and the use of hospitals that predominantly serve poor people ("safety net hospitals"). Quasi-experimental difference in differences study examining change after reform in the share of procedures performed in safety net hospitals by race/ethnicity and area income, with adjustment for patients' residence, demographics, and comorbidity. State of Massachusetts, United States. Massachusetts residents aged 40-64 as the target beneficiaries of reform and similarly aged residents of New Jersey, New York, and Pennsylvania as the comparison (control) population. Number of knee and hip replacement procedures per 10 000 population and use of safety net hospitals. Procedure counts from state discharge data for 2.5 years before and after reform, and multivariate difference in differences. Poisson regression was used to adjust for demographics, economic conditions, secular time, and geographic factors to estimate the change in procedure rate associated with health reform by race/ethnicity and area income. Before reform, the number of procedures (/10 000) in Massachusetts was lower among Hispanic people (12.9, P<0.001) than black people (28.1) and white people (30.1). Overall, procedure use increased 22.4% during the 2.5 years after insurance expansion; reform in Massachusetts was associated with a 4.7% increase. The increase associated with reform was significantly higher among Hispanic people (37.9%, P<0.001) and black people (11.4%, P<0.05) than among white people (2.8%). Lower income was not associated with larger increases in procedure use. The share of knee and hip replacement procedures performed in safety net hospitals in Massachusetts decreased by 1.0% from a level of 12.7% before reform. The reduction was larger among Hispanic people (-6.4%, P<0.001) than white people (-1.0%), and among low income residents (-3.9%, p<0.001) than high income residents (0%). Insurance expansion can help reduce disparities by race/ethnicity but not by income in access to elective surgical care and could shift some elective surgical care away from safety net hospitals.


Behavior studies by capacitance sensing.
The movements of small burrowing animals submerged in sand are monitored through capacitance changes which detune radio-frequency oscillators. A new dimension of reptilian activity studies is possible by using this technique. Some early observations of several reptile species maintained in darkness in isothermal sand are given. Patterns of behavior during active phases, depth and rate of diving, and duration of quiescent periods are being revealed in detail for the first time.


Determinants for DNA target structure selectivity of the human LINE-1 retrotransposon endonuclease.
The human LINE-1 endonuclease (L1-EN) is the targeting endonuclease encoded by the human LINE-1 (L1) retrotransposon. L1-EN guides the genomic integration of new L1 and Alu elements that presently account for approximately 28% of the human genome. L1-EN bears considerable technological interest, because its target selectivity may ultimately be engineered to allow the site-specific integration of DNA into defined genomic locations. Based on the crystal structure, we generated L1-EN mutants to analyze and manipulate DNA target site recognition. Crystal structures and their dynamic and functional analysis show entire loop grafts to be feasible, resulting in altered specificity, while individual point mutations do not change the nicking pattern of L1-EN. Structural parameters of the DNA target seem more important for recognition than the nucleotide sequence, and nicking profiles on DNA oligonucleotides in vitro are less well defined than the respective integration site consensus in vivo. This suggests that additional factors other than the DNA nicking specificity of L1-EN contribute to the targeted integration of non-LTR retrotransposons.