AU040320 deficiency leads to disruption of acrosome biogenesis and infertility in homozygous mutant mice.
Study of knockout (KO) mice has helped understand the link between many genes/proteins and human diseases. Identification of infertile KO mice provides valuable tools to characterize the molecular mechanisms underlying gamete formation. The KIAA0319L gene has been described to have a putative association with dyslexia; surprisingly, we observed that homozygous KO males for AU040320, KIAA0319L ortholog, are infertile and present a globozoospermia-like phenotype. Mutant spermatozoa are mostly immotile and display a malformed roundish head with no acrosome. In round spermatids, proacrosomal vesicles accumulate close to the acroplaxome but fail to coalesce into a single acrosomal vesicle. In wild-type mice AU040320 localises to the trans-Golgi-Network of germ cells but cannot be detected in mature acrosomes. Our results suggest AU040320 may be necessary for the normal formation of proacrosomal vesicles or the recruitment of cargo proteins required for downstream events leading to acrosomal fusion. Mutations in KIAA0319L could lead to human infertility; we screened for KIAA0319L mutations in a selected cohort of globozoospermia patients in which no genetic abnormalities have been previously identified, but detected no pathogenic changes in this particular cohort.


Interleukin-13 displaying retargeted oncolytic measles virus strains have significant activity against gliomas with improved specificity.
The majority of glioblastoma multiforme (GBM) tumors (80%) overexpress interleukin-13 receptor alpha2 (IL-13Ralpha2), but there is no expression of IL-13Ralpha2 in normal brain. Vaccine strains of measles virus have significant antitumor activity against gliomas. We tested the hypothesis that measles virus entry could be retargeted via the IL-13Ralpha2. MV-GFP-H(AA)-IL-13 was generated from the Edmonston-NSe vaccine strain, by displaying human IL-13 at the C-terminus of the H protein, and introducing CD46 and signaling lymphocyte activation molecule (SLAM)-ablating mutations in H. The IL-13 retargeted virus showed significant cytopathic effect (CPE) against IL-13Ralpha2 overexpressing glioma lines, and lack of CPE/viral replication in normal human astrocytes and normal human fibroblasts not expressing IL-13Ralpha2. In vivo treatment of orthotopically implanted GBM12 xenografts demonstrated significant prolongation of survival in mice treated with the retargeted strain (P < 0.0001), and comparable activity between the IL-13R retargeted strain and MV-GFP (P = 0.6377). In contrast to MV-GFP-treated mice, administration of the retargeted strain in the central nervous system of measles replication-permissive Ifnar(ko) CD46 Ge mice resulted in lack of neurotoxicity. Strains of measles virus retargeted against the glioma-specific IL-13Ralpha2 receptor have comparable therapeutic efficacy, and improved specificity as compared with the unmodified measles virus strain MV-GFP in vitro and in vivo.


Effect of dietary copper source on response to coliform mastitis in dairy cows.
The effect of organic or inorganic dietary Cu on Escherichia coli mastitis was investigated in first-lactation heifers. Twenty-eight primigravid Holstein heifers were assigned to 3 treatments in a completely randomized block design with 10 blocks of 3 animals grouped by expected calving date. Treatments were as follows: basal diet [7.1 mg Cu/kg of dry matter (DM); CON] and diets supplemented with Cu (10 mg/kg of DM) as Cu sulfate (CUS) or as Cu proteinate (CUP). Treatments were fed individually from 60 d prepartum through 49 d of lactation. All heifers were marginally deficient at the onset of the experiment (liver Cu of 60 mg/kg) and did not differ between groups. Mean liver Cu concentrations were about 3-fold greater in CUS and CUP compared with CON at d 0, 21, and 42 of lactation. At d 34 postpartum, one pathogen-free quarter per cow was infused with Escherichia coli strain 727. Copper supplementation did not lower peak responses to challenge; however, CUP tended to offer some benefits: milk bacterial count with CUP was lower compared with CON at 24, 48, and 72 h and lower than CUS at 24 and 96 h, and postchallenge milk production tended to be greater for CUP. Clinical udder score was lower at 12 h for CUP and CUS compared with CON, and at 144 h CUP had lower clinical scores compared with CUS or CON. Somatic cell count, dry matter intake, plasma Cu, and plasma ceruloplasmin did not differ between treatments. Compared with the control diet or Cu sulfate supplement, supplementation with Cu proteinate tended to improve the clinical status of cows after live E. coli intramammary challenge.


Urolithin A attenuated ox-LDL-induced cholesterol accumulation in macrophages partly through regulating miR-33a and ERK/AMPK/SREBP1 signaling pathways.
Promoting cholesterol efflux from foam cells represents one of the therapeutic strategies for ameliorating atherosclerosis. Urolithin A (UA) has been shown before to attenuate ox-LDL induced endothelial dysfunction in endothelial cells with its anti-inflammatory properties. The aim of this study was to investigate whether UA could promote cholesterol efflux via modulating related microRNA (miR) and signaling pathways. RAW264.7 cells were treated with 50 μg mL-1 ox-LDL to induce foam cell formation. After treatment with UA at different concentrations, intercellular and extracellular cholesterol levels were determined. Expression of Erk1/2, AMPKα and their phosphorylation forms, and SREBP1, was analyzed by western-blotting. The effect of UA on miR-33a expression and the involvement of miR-33a in cholesterol efflux regulation were also investigated. UA reduced ox-LDL induced cholesterol accumulation in macrophage cells and promoted cholesterol efflux from cells. Compared with ox-LDL treated cells, UA treatment reduced the level of phosphorylated ERK1/2, increased the expression of phosphorylated AMPKα and decreased the SREBP1 expression. Moreover, UA decreased the miR-33a expression at the transcriptional level but increased the transcriptional expression of ATP-binding cassette transporter A1 (ABCA1) and ABCG1, two genes contributing to reverse cholesterol transport. Furthermore, pre-miR-33a attenuated cholesterol efflux induced by UA. Collectively, UA promoted the reverse cholesterol transport in macrophage-derived foam cells and interfered with cholesterol metabolism possibly through regulating the miRNA-33 expression and interaction with the ERK/AMPKα/SREBP1 signaling pathway.


Molecular analysis of wild and domestic sheep questions current nomenclature and provides evidence for domestication from two different subspecies.
Complete mitochondrial DNA (mtDNA) control regions (CR) were sequenced and analysed in order to investigate wild sheep taxonomy and the origin of domestic sheep (Ovis aries). The dataset for phylogenetic analyses includes 63 unique CR sequences from wild sheep of the mouflon (O. musimon, O. orientalis), urial (O. vignei), argali (O. ammon) and bighorn (O. canadensis) groups, and from domestic sheep of Asia, Europe and New Zealand. Domestic sheep occurred in two clearly separated branches with mouflon (O. musimon) mixed into one of the domestic sheep clusters. Genetic distances and molecular datings based on O. canadensis CR and mtDNA protein-coding sequences provide strong evidence for domestications from two mouflon subspecies. Other wild sheep sequences are in two additional well-separated branches. Ovis ammon collium and O. ammon nigrimontana are joined with a specimen from the transkaspian Ust-Urt plateau currently named O. vignei arkal. Ovis ammon ammon, O. ammon darwini and O. vignei bochariensis represent a separate clade and the earliest divergence from the mouflon group. Therefore, O. musimon, O. vignei bochariensis and Ust-Urt sheep are not members of a 'moufloniform' or O. orientalis species, but belong to different clades. Furthermore, Ust-Urt sheep could be a hybrid population or an O. ammon subspecies closely related to O. ammon nigrimontana.


Exploring prior diseases associated with incident late-onset Alzheimer's disease dementia.
Studies have identified prior conditions associated with late-onset Alzheimer's disease dementia (LOAD), but all prior diseases have rarely been screened simultaneously in the literature. Our objective in the present study was to identify prior conditions associated with LOAD and construct pathways for them. We conducted a population-based matched case-control study based on data collected in the National Health Insurance Research database of Taiwan and the Catastrophic Illness Certificate database for the years 1997-2013. Prior diseases definitions were based on the first three digits of the codes listed in the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM). Inclusion criteria required that each ICD-code existed for at least 1 year and incurred at least 2 outpatient visits or inpatient diagnosis. The case group comprised 4,600 patients newly diagnosed with LOAD in 2007-2013. The LOAD patients were matched by sex and age to obtain 4,600 controls. Using stepwise multivariate logistic regression analysis, diseases were screened for 1, 2 …, 9 years prior to the first diagnosis of LOAD. Path analysis was used to construct pathways between prior diseases and LOAD. Our results revealed that the following conditions were positively associated with the incidence of LOAD: anxiety (ICD-code 300), functional digestive disorder (ICD code 564), psychopathology-specific symptoms (ICD-code 307), disorders of the vestibular system (ICD-code 386), concussion (ICD-code 850), disorders of the urethra and urinary tract (ICD-code 599), disorders of refraction and accommodation (ICD-code 367), and hearing loss (ICD-code 389). A number of the prior diseases have previously been described in the literature in a manner identical to that in the present study. Our study supports the assertion that mental, hearing, vestibular system, and functional digestive disorders may play an important role in the pathogenesis of LOAD.


Template-switching during DNA synthesis by Thermus aquaticus DNA polymerase I.
Recombinant DNA molecules are often generated during the polymerase chain reaction (PCR) when partially homologous templates are available [e.g., see Pääbo et al. (1990) J. Biol. Chem. 265, 4718-4721]. It has been suggested that these recombinant molecules are a consequence of truncated extension products annealing to partially homologous templates on subsequent PCR cycles. However, we demonstrate here that recombinants can be generated during a single round of primer extension in the absence of subsequent heat denaturation, indicating that template-switching produces some of these recombinant molecules. Two types of template-switches were observed: (i) switches to pre-existing templates and (ii) switches to the complementary nascent strand. Recombination is reduced several fold when the complementary template strands are physically separated by attachment to streptavidin magnetic beads. This result supports the hypothesis that either the polymerase or at least one of the two extending strands switches templates during DNA synthesis and that interaction between the complementary template strands is necessary for efficient template-switching.


Plasma nerve growth factor (NGF) and inflammatory cytokines (IL-6 and MCP-1) in young and adult subjects with Down syndrome: an interesting pathway.
Down's syndrome (DS) is the most frequent chromosomal aberration in men and it is invariably associated with mental retardation. Plasma levels of nerve growth factor (NGF), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) from non demented DS subjects of three different age-cohorts (2-14 years; 20-50 yrs; >60 yrs) and healthy controls were measured. No clinical and sub-clinical inflammation was apparent in DS patients. Plasma levels of NGF were higher in children, adult and old DS subjects than in controls. However, a significant age-related decrease of NGF levels was present in DS subjects. Serum levels of IL-6 and MCP-1 were also increased in DS children and adults, but not in older DS patients. High levels of circulating NGF might protect DS from clinical complications of atherosclerosis. However, the striking decrement of peripheral NGF levels with advancing age may predispose DS to clinical manifestation of dementia after adulthood.


Suppression of phase-induced intensity noise in fibre optic delay line signal processors using an optical phase modulation technique.
A technique that can suppress the dominant phase-induced intensity noise in fibre optic delay line signal processors is presented. It is based on phase modulation of the optical carrier to distribute the phase noise at the information band into a high frequency band which can be filtered out. This technique is suitable for suppressing the phase noise in various delay line structures and for integrating in the conventional fibre optic links. It can also suppress the coherent interference effect at the same time. A model for predicting the amount of phase noise reduction in various delay line structures using the optical phase modulation technique is presented for the first time and is experimentally verified. Experimental results demonstrate the technique can achieve a large phase noise reduction in various fibre optic delay line signal processors.


Cog5-Cog7 crystal structure reveals interactions essential for the function of a multisubunit tethering complex.
The conserved oligomeric Golgi (COG) complex is required, along with SNARE and Sec1/Munc18 (SM) proteins, for vesicle docking and fusion at the Golgi. COG, like other multisubunit tethering complexes (MTCs), is thought to function as a scaffold and/or chaperone to direct the assembly of productive SNARE complexes at the sites of membrane fusion. Reflecting this essential role, mutations in the COG complex can cause congenital disorders of glycosylation. A deeper understanding of COG function and dysfunction will likely depend on elucidating its molecular structure. Despite some progress toward this goal, including EM studies of COG lobe A (subunits 1-4) and higher-resolution structures of portions of Cog2 and Cog4, the structures of COG's eight subunits and the principles governing their assembly are mostly unknown. Here, we report the crystal structure of a complex between two lobe B subunits, Cog5 and Cog7. The structure reveals that Cog5 is a member of the complexes associated with tethering containing helical rods (CATCHR) fold family, with homology to subunits of other MTCs including the Dsl1, exocyst, and Golgi-associated retrograde protein (GARP) complexes. The Cog5-Cog7 interaction is analyzed in relation to the Dsl1 complex, the only other CATCHR-family MTC for which subunit interactions have been characterized in detail. Biochemical and functional studies validate the physiological relevance of the observed Cog5-Cog7 interface, indicate that it is conserved from yeast to humans, and demonstrate that its disruption in human cells causes defects in trafficking and glycosylation.


A general HIV incidence inference scheme based on likelihood of individual level data and a population renewal equation.
We derive a new method to estimate the age specific incidence of an infection with a differential mortality, using individual level infection status data from successive surveys. The method consists of a) an SI-type model to express the incidence rate in terms of the prevalence and its derivatives as well as the difference in mortality rate, and b) a maximum likelihood approach to estimate the prevalence and its derivatives. Estimates can in principle be obtained for any chosen age and time, and no particular assumptions are made about the epidemiological or demographic context. This is in contrast with earlier methods for estimating incidence from prevalence data, which work with aggregated data, and the aggregated effect of demographic and epidemiological rates over the time interval between prevalence surveys. Numerical simulation of HIV epidemics, under the presumption of known excess mortality due to infection, shows improved control of bias and variance, compared to previous methods. Our analysis motivates for a) effort to be applied to obtain accurate estimates of excess mortality rates as a function of age and time among HIV infected individuals and b) use of individual level rather than aggregated data in order to estimate HIV incidence rates at times between two prevalence surveys.


Stochastic ion channel gating in dendritic neurons: morphology dependence and probabilistic synaptic activation of dendritic spikes.
Neuronal activity is mediated through changes in the probability of stochastic transitions between open and closed states of ion channels. While differences in morphology define neuronal cell types and may underlie neurological disorders, very little is known about influences of stochastic ion channel gating in neurons with complex morphology. We introduce and validate new computational tools that enable efficient generation and simulation of models containing stochastic ion channels distributed across dendritic and axonal membranes. Comparison of five morphologically distinct neuronal cell types reveals that when all simulated neurons contain identical densities of stochastic ion channels, the amplitude of stochastic membrane potential fluctuations differs between cell types and depends on sub-cellular location. For typical neurons, the amplitude of membrane potential fluctuations depends on channel kinetics as well as open probability. Using a detailed model of a hippocampal CA1 pyramidal neuron, we show that when intrinsic ion channels gate stochastically, the probability of initiation of dendritic or somatic spikes by dendritic synaptic input varies continuously between zero and one, whereas when ion channels gate deterministically, the probability is either zero or one. At physiological firing rates, stochastic gating of dendritic ion channels almost completely accounts for probabilistic somatic and dendritic spikes generated by the fully stochastic model. These results suggest that the consequences of stochastic ion channel gating differ globally between neuronal cell-types and locally between neuronal compartments. Whereas dendritic neurons are often assumed to behave deterministically, our simulations suggest that a direct consequence of stochastic gating of intrinsic ion channels is that spike output may instead be a probabilistic function of patterns of synaptic input to dendrites.


Left Atrial Remodeling and Atrioventricular Coupling in a Canine Model of Early Heart Failure With Preserved Ejection Fraction.
Left atrial (LA) compliance and contractility influence left ventricular stroke volume. We hypothesized that diminished LA compliance and contractile function occur early during the development of heart failure with preserved ejection fraction (HFpEF) and impair overall cardiac performance. Cardiac magnetic resonance imaging, echocardiography, left ventricular and LA pressure-volume studies, and tissue analyses were performed in a model of early HFpEF (elderly dogs, renal wrap-induced hypertension, exogenous aldosterone; n=9) and young control dogs (sham surgery; n=13). Early HFpEF was associated with LA enlargement, cardiomyocyte hypertrophy, and enhanced LA contractile function (median active emptying fraction 16% [95% confidence interval, 13-24]% versus 12 [10-14]%, P=0.008; end-systolic pressure-volume relationship slope 2.4 [1.9-3.2]mm Hg/mL HFpEF versus 1.5 [1.2-2.2]mm Hg/mL controls, P=0.01). However, atrioventricular coupling was impaired and the curvilinear LA end-reservoir pressure-volume relationship was shifted upward/leftward in HFpEF (LA stiffness constant [βLA] 0.16 [0.11-0.18]mm Hg/mL versus 0.06 [0.04-0.10]mm Hg/mL controls; P=0.002), indicating reduced LA compliance. Impaired atrioventricular coupling and lower LA compliance correlated with lower left ventricular stroke volume. Total fibrosis and titin isoform composition were similar between groups; however, titin was hyperphosphorylated in HFpEF and correlated with βLA. LA microvascular reactivity was diminished in HFpEF versus controls. LA microvascular density tended to be lower in HFpEF and inversely correlated with βLA. In early-stage hypertensive HFpEF, LA cardiomyocyte hypertrophy, titin hyperphosphorylation, and microvascular dysfunction occur in association with increased systolic and diastolic LA chamber stiffness, impaired atrioventricular coupling, and decreased left ventricular stroke volume. These data indicate that maladaptive LA remodeling occurs early during HFpEF development, supporting a concept of global myocardial remodeling.


Inhibition of Trigeminal Nociception by Non-invasive Vagus Nerve Stimulation: Investigating the Role of GABAergic and Serotonergic Pathways in a Model of Episodic Migraine.
Migraine is a prevalent neurological disease that is characterized by unpredictable episodic attacks of intense head pain. The underlying pathology involves sensitization and activation of the trigeminal system. Although non-invasive vagus nerve stimulation (nVNS) is recommended for the treatment of migraine, the abortive mechanism of action is not well-understood. The goal of this study was to compare the ability of nVNS and sumatriptan to inhibit trigeminal activation in two animal models of episodic migraine and to investigate the receptor mechanism of action of nVNS. Nocifensive head withdrawal response was investigated in adult male Sprague Dawley rats using von Frey filaments. To induce trigeminal nociceptor sensitization, complete Freund's adjuvant was injected in the trapezius muscle and trigeminal neurons were activated by exposure to a pungent odor or injection of the nitric oxide donor sodium nitroprusside. Some animals received nVNS or sumatriptan as treatment. Some animals were injected intracisternally with antagonists of GABAA, 5-HT3 or 5-HT7 receptors prior to nVNS since these receptors are implicated in descending modulation. While unsensitized animals exposed to the pungent odor or nitric oxide alone did not exhibit enhanced mechanical nociception, sensitized animals with neck muscle inflammation displayed increased trigeminal nocifensive responses. The enhanced nociceptive response to both stimuli was attenuated by nVNS and sumatriptan. Administration of antagonists of GABAA, 5-HT3, and 5-HT7 receptors in the upper spinal cord suppressed the anti-nocifensive effect of nVNS. Our findings suggest that nVNS inhibits trigeminal activation to a similar degree as sumatriptan in episodic migraine models via involvement of GABAergic and serotonergic signaling to enhance central descending pain modulation.


ZAP-70+ B cell subset influences response to B cell depletion therapy and early repopulation in rheumatoid arthritis.
To define the role of ZAP-70+ B cells (CD19+/ZAP-70+) as a biomarker of response to B cell depletion therapy (BCDT), their relationship with clinical outcome, and their behavior during repopulation of peripheral blood in patients with rheumatoid arthritis (RA). Thirty-one patients with RA underwent BCDT and were followed for 12 months. Disease activity was assessed with the European League Against Rheumatism (EULAR) criteria. Cytofluorimetric analysis of peripheral blood B cell subsets at baseline and at 6- and 12-month intervals after BCDT was performed using surface markers (CD45, CD3, CD56, CD19, IgD, CD38, CD27) and intracellular ZAP-70. A moderate/good EULAR response was achieved in 66.6% of the RA cohort. The baseline percentage of CD19+/ZAP-70+ cells was lower in good responder patients (1.8% ± 1.7%) compared to poor responders (5.6% ± 4.9%; p = 0.02). A decrease of plasmablasts (IgD-CD27+CD38+) and pre-switch memory (IgD+CD27+) B cells occurred after BCDT. Recovery of B cells in peripheral blood after the first course of BCDT was characterized by the reappearance of B cell subtypes that showed a naive, activated phenotype, coupled with a decrease in memory cells. B cells carrying intracytoplasmic ZAP-70 increased significantly from the baseline value of 4.4% ± 4.5% to 12.4% ± 9.2% (p = 0.001) at the 6-month and to 9.4% ± 6.4% (p = 0.002) at the 12-month followup. Baseline percentage of CD19+/ZAP-70+ cells is associated with the clinical outcome after BCDT in patients with RA. Depletion of plasmablasts and pre-switch memory B cells and increase of CD19+/ZAP-70+ cells are features of the recovery of the B cell pool after BCDT.


Ratings of Perceived Exertion and Physiological Parameters of Muscle Metabolism in Postmenopausal Women.
We compared responses from postmenopausal women living a sedentary lifestyle ( n = 15; Mean age= 59; SD = 4.2) to a single bout of water- or land-based exercise with respect to ratings of perceived exertion (RPE), lactate concentration, and muscle oxygen saturation. Each participant was randomly assigned to a single water- or land-based 50-minute bout of combined aerobic and resistance exercise. Blood samples were collected to detect pre- and post-exercise lactate concentration. Total hemoglobin, deoxidized hemoglobin, and the percentage change in the total oxygen saturation index (TSI%) of the rectus femoris were detected by means of near-infrared spectroscopy. We found similar RPE at various stages of land- and water-based exercise, and a similar change in lactate concentration in these environments (in water: 4.35 ± 1.49 mol/L; on land: 3.62 ± 1.18 mol/L). However, the reduction in HHb response was less pronounced after water-based exercise, and TSI% increased on land but decreased in water, with the magnitude of this change much higher on land. For similar RPE and lactate concentration, the oxygen saturation in the exercising muscles decreased in water, suggesting higher oxygen consumption in water than on land.


Prostaglandin E1 effects on CD62p and PAC-1 in patients with sudden sensorineural hearing loss.
To evaluate the treatment outcome of vasodilator prostaglandin E1 (PGE1) in treating sudden sensorineural hearing loss (SSNHL) and to determine its effects on platelet activation, as reflected by changes in CD62p and PAC-1. We prospectively enrolled 60 patients with confirmed SSNHL and randomly divided them into two groups: the SSNHL group received regular therapy, and the SSNHL-PGE1 group received additional intravenous injection of PGE1. After 14 days of treatment, we measured clinical improvement and CD62p-positive and PAC-1-positive platelets. 30 healthy medical staff members were included as a control group. The SSNHL patients had significantly higher levels of CD62p-positive or PAC-1 positive platelets than the healthy subjects. The ratios of CD62p positive or PAC-1 positive platelets significantly decreased after the two treatments. The average pure tone (PTA) hearing thresholds decreased to 26.51 ± 12.65 dB in SSNHL-PGE1 group after treatment, which was significantly lower than that of the SSNHL group (34.46 ± 10.35 dB). Patients with initial severe or profound hearing loss (PTA ≥ 71 dB) had better hearing improvement on PGE1 than on the regular treatment. Patients in the SSNHL-PGE1 treatment group had significantly lower CD62p and PAC-1 levels than those in the SSNHL group. Patients with higher initial positive CD62p and PAC-1 ratios tended to have higher potential of clinical improvement and hearing gains after PGE1 treatment. Initial CD62p and PAC-1 levels were significantly correlated with hearing thresholds in patients with SSNHL. PGE1 application could improve treatment efficacy and suppress excessive platelet activation in patients with SSNHL.


Associations of Physical Activity and Sedentary Behavior with Atopic Disease in United States Children.
To determine if eczema, asthma, and hay fever are associated with vigorous physical activity, television/video game usage, and sports participation and if sleep disturbance modifies such associations. Data were analyzed from 2 cross-sectional studies including 133 107 children age 6-17 years enrolled in the 2003-2004 and 2007-2008 National Survey of Children's Health. Bivariate and multivariate survey logistic regression models were created to calculate the odds of atopic disease and atopic disease severity on vigorous physical activity, television/video game use, and sports participation. In multivariate logistic regression models controlling for sociodemographic factors, lifetime history of asthma was associated with decreased odds of ≥1 days of vigorous physical activity (aOR, 0.87; 95% CI, 0.77-0.99) and decreased odds of sports participation (0.91; 95% CI, 0.84-0.99). Atopic disease accompanied by sleep disturbance had significantly higher odds of screen time and lower odds of sports participation compared with children with either atopic disease or sleep disturbance alone. Severe eczema (aOR, 0.39; 95% CI, 0.19-0.78), asthma (aOR, 0.29; 95% CI, 0.14-0.61), and hay fever (aOR, 0.48; 95% CI, 0.24-0.97) were all associated with decreased odds of ≥1 days of vigorous physical activity. Moderate (aOR, 0.76; 95% CI, 0.57-0.99) and severe eczema (aOR, 0.45; 95% CI, 0.28-0.73), severe asthma (aOR, 0.47; 95% CI, 0.25-0.89), and hay fever (aOR, 0.53; 95% CI, 0.36-0.61) were associated with decreased odds of sports participation in the past year. Children with severe atopic disease, accompanied by sleep disturbance, have higher risk of sedentary behaviors.


[Pharmaceutical assistance in the Brazilian public health care system].
This article deals with the organization of pharmaceutical assistance within the Brazilian Public Unified Health System (Sistema Unico de Saúde, SUS) by analyzing the progress made so far and the challenges that still must be faced to ensure the right to universal therapeutic and pharmaceutical coverage in Brazil. Among the improvements, the article discusses the National List of Essential Drugs and its role to guide the supply, prescription, and dispensing of drugs within SUS. The structure of federal funding and the existing pharmaceutical programs are described, as well as the responsibilities of the Federal Government, states, and municipalities concerning allocation of financial resources. Despite the progress made, the model of access to SUS drugs has not been successful in securing universal therapeutic assistance. It is essential to consider a type of funding and organization of pharmaceutical assistance that is driven by a notion of regionalization, with financing based on health care flows rather than on the population circumscribed to a given territory. It is also essential to qualify pharmaceutical management by hiring pharmacists and support personnel, professionalizing workers, and setting up facilities with appropriate information systems and equipment.


Effect of surgeon and hospital characteristics on outcome after pyloromyotomy.
Previous studies have suggested that the outcome after pyloromyotomy is improved with increased surgeon experience. Others have proposed that infants with pyloric stenosis are best treated by specialty-trained pediatric surgeons or at children's hospitals. Surgeon and hospital characteristics affect complications, length of stay, and hospital charges after pyloromyotomy. Data for a nationally representative sample of infants (n = 1277) who underwent pyloromyotomy in 2000 in the United States were obtained from the Kids' Inpatient Database. Surgeon and hospital volumes were stratified into quintiles. Multivariate analyses were performed to analyze the impact of surgeon and hospital volume on length of stay, charges, and major operative complications using models that accounted for the hierarchical structure of patient-, surgeon-, and hospital-level covariates. No association between surgeon volume and either length of stay or charges was observed. Higher surgeon volume, however, was associated with fewer complications (P<.001). Surgeons with the highest volume had a 90% lower risk of complications than those with the lowest volume. Higher hospital volume was associated with shorter length of stay (P<.001). No association between hospital volume and either charges or risk of complications was observed. Higher surgeon and hospital volumes are associated with better outcome among infants who are treated for pyloric stenosis. Identification of aspects of medical and surgical treatment that account for this finding may lead to improvement in the outcome of infants undergoing pyloromyotomy.


Use of p53-Silenced Endothelial Progenitor Cells to Treat Ischemia in Diabetic Peripheral Vascular Disease.
Peripheral vascular disease is a major diabetes mellitus-related complication. In this study, we noted that expressions of proapoptotic p53 gene and its downstream cascade gene such as p21 are upregulated in hyperglycemia. Therefore, we investigated whether p53- and p21-silenced endothelial progenitor cells (EPCs) were able to survive in hyperglycemic milieu, and whether transplantation of either p53 knockout (KO) or p21KO or p53- and p21-silenced EPCs could improve collateral vessel formation and blood flow in diabetic vaso-occlusive peripheral vascular disease mouse models. We transplanted p53 and p21KO mouse EPCs (mEPCs) into streptozotocin-induced diabetic (type 1 diabetes mellitus model) C57BL/6J and db/db (B6.BKS(D)-Leprdb/J) (type 2 model) post-femoral artery occlusion. Similarly, Ad-p53-silenced and Ad-p21-silenced human EPCs (CD34+) cells were transplanted into streptozotocin-induced diabetic NOD.CB17-Prkdcscid/J mice. We measured blood flow at 3, 7, and 10 days and hindlimb muscles were obtained postsacrifice for mRNA estimation and CD31 staining. Enhanced blood flow was noted with delivery of p53 and p21KO mEPCs in streptozotocin-induced diabetic C57BL/6J mice. Similar results were obtained when human Ad-p53shEPCs(CD34+) and Ad-p21shEPCs(CD34+) were transplanted into streptozotocin-induced nonobese diabetic severe combined immunodeficiency mice. Gene expression analysis of p53 and p21KO EPCs transplanted hindlimb muscles showed increased expression of endothelial markers such as endothelial nitric oxide synthase, vascular endothelial growth factor A, and platelet endothelial cell adhesion molecule 1. Similarly, quantitative reverse transcriptase polymerase chain reaction of human Ad-p53shEPCs (CD34+)- and Ad-p21shEPCs (CD34+)-transplanted hindlimb muscles also showed increased expression of endothelial markers such as vascular endothelial growth factor A, noted primarily in the p53-silenced EPCs group. However, such beneficial effect was not noted in the db/db type 2 diabetic mouse models. Transient silencing of p53 using adenoviral vector in EPCs may have a therapeutic role in diabetic peripheral vascular disease.


Metabolic Adaptations of CD4+ T Cells in Inflammatory Disease.
A controlled and self-limiting inflammatory reaction generally results in removal of the injurious agent and repair of the damaged tissue. However, in chronic inflammation, immune responses become dysregulated and prolonged, leading to tissue destruction. The role of metabolic reprogramming in orchestrating appropriate immune responses has gained increasing attention in recent years. Proliferation and differentiation of the T cell subsets that are needed to address homeostatic imbalance is accompanied by a series of metabolic adaptations, as T cells traveling from nutrient-rich secondary lymphoid tissues to sites of inflammation experience a dramatic shift in microenvironment conditions. How T cells integrate information about the local environment, such as nutrient availability or oxygen levels, and transfer these signals to functional pathways remains to be fully understood. In this review, we discuss how distinct subsets of CD4+ T cells metabolically adapt to the conditions of inflammation and whether these insights may pave the way to new treatments for human inflammatory diseases.


Unique features of the mode of action of ET-743.
This paper describes the current knowledge of the primary mode of action of a natural product, ecteinascidin 743 (ET-743), derived from the marine tunicate Ecteinascidia turbinata. ET-743 was initially selected for preclinical development because of its potent antitumor activity observed against several human solid tumor types. In vitro, the drug is cytotoxic in the nanomolar range, and in the case of some very sensitive cell lines, in the picomolar range. The large potency differences observed among several solid tumor types indicate that this compound possesses some tumor selectivity, but the molecular basis of these differential effects remains to be elucidated. The present studies were undertaken to evaluate the mechanism of action of ET-743 in this context. The available information on ET-743 binding to DNA and its effects on transcriptional regulation point to a unique behavior of this drug, as it independently affects specific gene transcription in a promoter-dependent way. In addition, ET-743 shows a peculiar pattern of selectivity in cells with different defects in their DNA-repair pathways. These results highlight a unique property of ET-743, possibly explaining why it possesses antitumor activity against tumors that are refractory to standard anticancer drugs, all of which certainly act by mechanisms that are different from that of ET-743.


Proposal for standardized definitions for efficacy end points in adjuvant breast cancer trials: the STEEP system.
Standardized definitions of breast cancer clinical trial end points must be adopted to permit the consistent interpretation and analysis of breast cancer clinical trials and to facilitate cross-trial comparisons and meta-analyses. Standardizing terms will allow for uniformity in data collection across studies, which will optimize clinical trial utility and efficiency. A given end point term (eg, overall survival) used in a breast cancer trial should always encompass the same set of events (eg, death attributable to breast cancer, death attributable to cause other than breast cancer, death from unknown cause), and, in turn, each event within that end point should be commonly defined across end points and studies. A panel of experts in breast cancer clinical trials representing medical oncology, biostatistics, and correlative science convened to formulate standard definitions and address the confusion that nonstandard definitions of widely used end point terms for a breast cancer clinical trial can generate. We propose standard definitions for efficacy end points and events in early-stage adjuvant breast cancer clinical trials. In some cases, it is expected that the standard end points may not address a specific trial question, so that modified or customized end points would need to be prospectively defined and consistently used. The use of the proposed common end point definitions will facilitate interpretation of trial outcomes. This approach may be adopted to develop standard outcome definitions for use in trials involving other cancer sites.


Pyrrolidinium ionic liquid crystals with pendant mesogenic groups.
New ionic liquid crystals (including ionic metallomesogens) based on the pyrrolidinium core are presented. N-Methylpyrrolidine was quaternized with different mesogenic groups connected to a flexible, omega-bromosubstituted alkyl spacer. The length of the flexible alkyl spacer between the cationic head group and the rigid mesogenic group was varied. The substituted pyrrolidinium cations were combined with bromide, bis(trifluoromethylsulfonyl)imide, tetrakis(2-thenoyltrifluoroacetonato)europate(III), and tetrabromouranyl anions. The influence of the type of mesogenic unit, the lengths of the flexible spacer and terminal alkyl chain, the size of the mesogenic group, and the type of anion on the thermotropic mesomorphic behavior was investigated. Furthermore, the phase behavior was thoroughly compared with the previously reported mesomorphism of N-alkyl-N-methylpyrrolidinium salts. Low-ordered smectic A phases of the de Vries type, smectic C phases, higher-ordered smectic F/I phases, as well as crystal smectic phases (E and G, J, H, or K) were observed and investigated by polarizing optical microscopy, differential scanning calorimetry, and powder X-ray diffraction.


Differential mobilization of blubber fatty acids in lactating Weddell seals: evidence for selective use.
A major source of energy during lactation in mammals is provided through the mobilization of blubber fatty acids (FAs). We investigated the extent to which FAs were mobilized to support both maternal metabolic requirements and milk production in the Weddell seal and how this was reflected in the FA composition of the pup's blubber at the end of lactation (EL). FA composition of postpartum female blubber was similar in the 2 yr of study (2002 and 2003) but differed markedly by EL. Pup blubber FAs (at EL) were also different between years and did not match that of the mother's milk or blubber. Milk FA composition changed during lactation, which may have been a reflection of an increase in pup energy demands at different stages of development. In addition, there was evidence of feeding by some females during lactation, with higher levels of some FAs in the milk than in the blubber. Our results indicate that differential mobilization of FAs occurred in lactating Weddell seals and that this was related to total body lipid stores at postpartum. Furthermore, growing pups did not store FAs unmodified, providing evidence that selective use does occur and also that using FA composition to elucidate dietary sources may be problematic in growing individuals.


Structure and expression of the chicken calmodulin I gene.
The chicken calmodulin I (CaMI) gene has been isolated and characterized on the level of cDNA and genomic DNA. The deduced amino acid (aa) sequence is identical to the one of chicken CaMII which consists of 148 aa. The CaMI gene contains six exons. Its intron/exon organization is identical to that of the chicken CaMII and the CaMI and CaMIII genes of rat and human. Expression of the CaMI gene was detected in all chicken tissues examined, although at varying levels. The gene is transcribed into four mRNAs of 0.8, 1.4, 1.7 and 4.4 kb as determined by Northern blot analysis. Our results demonstrate that the "multigene-one-protein" principle of CaM synthesis is not only applicable to mammals whose CaM is encoded by three different genes, but also to chickens.


Genetic analysis of mammographic breast density in adult women: evidence of a gene effect.
The appearance of the female breast viewed by mammography varies considerably from one individual to another because of underlying differences in the relative proportions of fat, connective tissue, and glandular epithelium that combine to create a characteristic pattern of breast density. An association between mammographic patterns and family history of breast cancer has previously been reported. However, this association has not been found in all studies, and few data are available on possible genetic components contributing to mammographic breast density. Our purpose was to estimate familial correlations and perform complex genetic segregation analyses to test the hypothesis that the transmission of a major gene influences mammographic breast density. As part of a cohort study (initiated in 1944) of families with a history of breast cancer, the probands' female relatives who were older than 40 years were asked to obtain a routine mammogram. The mammograms of 1370 women from 258 independent families were analyzed. The fraction of the breast volume occupied by radiographically dense tissue was estimated visually from video displays of left or right mediolateral oblique views by one radiologist experienced in mammography who had no knowledge of individual relationships to the probands. Data on breast cancer risk factors were obtained through telephone interviews and mailed questionnaires. Unadjusted and adjusted familial correlations in breast density were calculated, and complex genetic segregation analyses were performed. Sister-sister correlations in breast density (unadjusted and adjusted for age and either body mass index, menopausal status, hormone replacement therapy, waist-to-hip ratio, number of live births, alcohol consumption, or cigarette smoking status) were all statistically significant (r = .16-.27; all P<.05 [two-sided]). Estimated mother-daughter correlations were smaller in magnitude (r = .01-.17) and not statistically significant. Segregation analyses indicate that a major autosomal gene influences breast density. The mendelian transmission of a dominant gene provided the best fit to the data; however, hypotheses involving the inheritance of either a recessive gene or a codominant gene could not be ruled out. The mendelian dominant hypothesis, accounting for 29% of the variability in breast density, suggests that approximately 12% of the population would be expected to carry at least one variant allele of this putative gene. Women who inherit the variant allele would have a mean breast density about twice that of the rest of the population. Our preliminary findings suggest that, in this cohort of women at risk of breast cancer, mammographic breast density may be genetically influenced.


Mortality patterns of Native Hawaiians across their lifespan: 1990-2000.
We examined mortality patterns across the lifespan of Native Hawaiians and compared mortality disparities across races. We determined the age-specific and age-adjusted mortality rates of Native Hawaiians from 1990 to 2000 by using national census and vital registration data. Among Native Hawaiians aged younger than 1 year, expected deaths were 15% lower than for Blacks and 50% higher than for Whites. Among older adults, Native Hawaiians had higher rates of mortality compared with the general population, particularly in 1990 and 1995. Crude death rates for Native Hawaiians were similar to those for Blacks in 1990 and 1995 but were 20% lower than those for Blacks by 2000. Crude death rates for Native Hawaiians were 30% higher than for Whites in 1990 and 1995 and more than 40% higher than for Whites in 2000. Compared with Whites, Native Hawaiians and Blacks face similar challenges regarding infant and early-life mortality and increasing risks of mortality in mid-life and early old age. Our analyses document a need for renewed efforts to identify the determinants of ill health and commitment to address them.


Sleep patterns of day-working, evening high-schooled adolescents of São Paulo, Brazil.
Children who grow up in developing countries of the world must work to help financially support their families, and they must also attend school. We investigated the impact of work on the sleep of working vs. nonworking high school students. Twenty-seven São Paulo, Brazil, public high school students (eight male and eight female working students plus six nonworking female and five nonworking male students) 14-18 yrs of age who attended school Monday-Friday between 19:00 to 22:30h participated. A comprehensive questionnaire about work and living conditions, health status, and diseases and their symptoms was also answered. The activity level and rest pattern (sleep at night and napping during the day) were continuously assessed by wrist actigraphy (Ambulatory Monitoring, USA). The main variables were analyzed by a two-factor ANOVA with application of the Tukey HSD test for multiple comparisons, and the length of sleep during weekdays vs. weekends was compared by Student t-test. Working students went to sleep earlier weekends [F(1,23)=6.1; p=0.02] and woke up earlier work days than nonworking students [F(1,23) = 17.3; p = 0.001]. The length of nighttime sleep during weekdays was shorter among all the working [F(1,23)= 16.7; p <0.001] than all the nonworking students. The sleep duration of boys was shorter than of girls during weekends [F(1,23)= 10.8; p <0.001]. During weekdays, the duration of napping by working and nonworking male students was shorter than nonworking female students. During weekdays working girls took the shortest naps [F(1,23)= 5.6; p = 0.03]. The most commonly reported sleep complaint during weekdays was difficulty waking up in the morning [F(1,23) = 6.5; p = 0.02]. During weekdays, the self-perceived sleep quality of working students was worse than nonworking students [F(1,23) = 6.2; p = 0.02]. The findings of this study show that work has negative effects on the sleep of adolescents, with the possible build-up of a chronic sleep debt with potential consequent impact on quality of life and school learning.


Sero-prevalence of granulocytic Ehrlichia spp. and Borrelia burgdorferi sensu lato in Swedish dogs 1991-94.
In Sweden, 2 tick-borne zoonotic diseases, granulocytic ehrlichiosis and borreliosis, are frequently diagnosed in dogs, using serological assays. The aims of this study were to determine the sero-prevalences of antibodies to Ehrlichia spp. and Borrelia burgdorferi sensu lato during 1991-94 in dogs, not clinically suspected to be infected with either of the 2 agents. Samples (n = 611) were selected from a serum bank using a systematic sampling strategy, stratified across the 4-y period. The stored sera had originally been submitted in order to verify or rule out infection with Sarcoptes scabiei. The overall sero-prevalence for Ehrlichia spp. was 17.7% and for Borrelia burgdorferi sensu lato 3.9% (n = 588). Only a few dogs in the northern part of Sweden were sero-positive for Ehrlichia spp. and none were positive for Borrelia burgdorferi sensu lato. An increased sero-prevalence of Ehrlichia spp. was seen during the years studied. The sero-prevalence of Ehrlichia spp. varied with season. Sero-positivity to both agents increased with age. Both diseases are considered zoonotic, and the increase in sero-prevalence of Ehrlichia spp. over the years may reflect the degree of infection in ticks and may have implications for human health.


Behavioral, neuroendocrine, and immunological outcomes of escapable or inescapable shocks.
The present study was designed to evaluate the effects of repeated exposure to escapable or inescapable shocks on subsequent behavior in an activity cage, and on the reactivity of the hypothalamic-pituitary-adrenocortical (HPA) axis and the immune system. We also studied the possible influence of behavioral factors on the behavioral and physiological impact of stress. Although exposure to different stressful situations pointed out marked differential effects in subsequent behavior, it failed to elicit differences in the neuroendocrine and immunological parameters studied. However, interesting results were found in analyzing the influence of behavioral factors. The degree of control exerted over the shock was inversely related to ACTH and corticosterone levels. In addition, individual differences in the exploratory activity to novelty were correlated with poststress lymphoproliferation and antibody formation. These data indicate that the behavioral and physiological outcomes of stress depend on the interrelations between environmental and individual factors (including both preexisting individual differences and the coping responses during stress).


Response of some common annual bedding plants to three species of meloidogyne.
Twelve ornamental bedding plant cultivars were grown in soil infested with isolates of Meloidogyne incognita race 1, M. javanica, or M. arenaria race 1 in a series of tests in containers in a growth room. Root galling (0-5 scale) and eggs/plant were evaluated 8-10 weeks after soil infestation and seedling transplantation. Snapdragon, Antirrhinum majus cv. First Ladies, was extensively galled and highly susceptible (mean gall rating >/=4.2 and >/=14,500 eggs/plant), and Celosia argentea cv. Century Mix and Coleus blumei cv. Rainbow were susceptible (>1,500 eggs/plant) to all three Meloidogyne isolates. Response of Petunia x hybrida varied with cultivar and nematode isolate. Little or no galling or egg production from any Meloidogyne isolate was observed on Ageratum houstonianum cv. Blue Mink, Lobularia maritima cv. Rosie O'Day, or Tagetes patula cv. Dwarf Primrose. Galling was slight (mean rating </=2.0) but varied with nematode species on Dianthus chinensis cv. Baby Doll Mix, Salvia splendens cv. Bonfire, and Vinca rosea cv. Little Bright Eye. Verbena x hybrida cv. Florist was heavily infected (gall rating >4.0 and >/=7,900 eggs/plant) by M. javanica and M. arenaria but was nearly free of galling from M. incognita. Zinna elegans cv. Scarlet was nearly free of galling from M. incognita and M. arenaria but was susceptible (mean gall rating = 2.9; 3,400 eggs/plant) to M. javanica.


Effects of secondary warm up following stretching.
Evidence suggests that static stretching inhibits muscular power. However, research does not reflect practice whereby individuals follow up stretching with secondary activity. This study investigated muscular power following stretching, and after a second bout of activity. Participants (n = 13) completed 3 randomized testing sessions which included a 5 min warm-up, followed by a vertical jump (VJ) on a force platform; an intervention (static stretching, dynamic, or control), followed by a second VJ. Participants then completed a series of movements, followed by a VJ, up to 60 min post activity. Immediately following the intervention, there was a 10.7% difference in VJ between static and dynamic stretching. The second warm up bout increased VJ height following the dynamic intervention, whereas the static stretching condition did not show any differences. The novel finding from this study demonstrates a second exercise bout does not reverse the effects of static stretching and is still detrimental to VJ.


Disseminated Mycobacterium haemophilum infection in a 72-year-old patient with rheumatoid arthritis on infliximab.
Mycobacterium haemophilum is a slow growing, aerobic, fastidious mycobacterium requiring hemin and a temperature of 30-32° C for optimal growth that is ubiquitous in nature. Disease in immunocompromised adults typically manifests as skin lesions such as papules, pustules and ulcerations. This organism also causes lymphadenitis in immunocompetent children. Antitumour necrosis factor-α (anti-TNF-α) therapy with agents such as infliximab, etanercept and adalimumab is increasingly being used for immunosuppression in patients with various autoimmune conditions. These agents are known to place patients at increased risk for tuberculosis and other granulomatous diseases. However, little is known about illness caused by M haemophilum in patients on immunosuppression with anti-TNF-α therapy. We describe a case of disseminated M haemophilum manifesting as skin lesions in a 72-year-old man with rheumatoid arthritis on infliximab and methotrexate.


Modeling of Human Operator Behavior for Brain-Actuated Mobile Robots Steering.
Human operator control of brain-actuated robot steering based on electroencephalograph (EEG)-signals is a complex behavior consisting of surroundings perceiving, decision making, and commands issuing and differs among individual operators. However, no existing models allow decoupling the user from the loop to improve the system design and testing process, which can capture such behavior of a brain-actuated robot. To address this problem, in this paper, we propose an operator brain-controlled steering model consisting of an operator decision model based on the queuing network (QN) cognitive architecture and a brain-machine interface (BMI) performance model. The QN-based operator decision model can mimic the human decision process with the individual operator differences considered. The new BMI performance model is built to represent the varied accuracy of BMI during brain-controlled direction operations. Furthermore, the model is simulated and validated against the results of human operator-in-the-loop experiments. The results show that the proposed model can reproduce the behavior of human operators thanks to its similar direction control performance.


Self-control and impulsiveness in children and adults: Effects of food preferences.
Experiment 1 used 6 preschool boys and Experiment 2 used 6 adult women to explore the effects of food preference on humans' choice in self-control paradigms. The boys showed a higher proportion of responses for more delayed, larger reinforcers (a measure of self-control) when those choices resulted in receipt of the most preferred food compared to when those choices resulted in the least preferred food. Further, the boys chose the less delayed, smaller reinforcers significantly more often when only those choices, as opposed to both choices, resulted in the most preferred food. Conversely, they chose the more delayed, larger reinforcers significantly more often when only those choices, as opposed to both choices, resulted in the most preferred food. Finally, the women demonstrated significantly less sensitivity to reinforcer amount relative to sensitivity to reinforcer delay (another measure of self-control) when they had a higher preference for the juice received as the less delayed, smaller reinforcer than for the juice received as the more delayed, larger reinforcer. Together, the results show that subjects' food preferences can influence self-control for food reinforcers.


Objectively Measured School Day Physical Activity Among Elementary Students in the United States and Finland.
Schools are in a unique position to ensure that all students meet the current physical activity (PA) recommendations. This study aimed to examine 1st to 3rd grade elementary students' accelerometer measured school day PA in the United States (U.S.) and Finland. The sample consisted of 200 students (107 girls, 93 boys; ages 6 to 8) and their school day PA was monitored with hip-worn ActiGraph GT3X+ accelerometers across a 5-day school week and the thresholds 100 and 2296 count per minute were used to separate sedentary time, light PA, and moderate-to-vigorous PA (MVPA). On an average school day, students were engaged in MVPA for 20.0 min in the U.S. and 24.1 min in Finland. Students' school-day MVPA was 9 to 16 minutes higher during physical education (PE) days compared with non-PE days (U.S: 25.8 vs. 16.6 min/day; Finland: 36.3 vs. 20.1 min/day). Girls had less MVPA and more sedentary time compared with boys in both samples. This study highlights both the role of PE and other school day physical activities in meeting PA guidelines. Policy measures are needed to change the structure of the school day and enhance PA to ensure that students meet the PA recommendations.


Prognostic factors for progression of osteoarthritis of the hip: a systematic review.
Predicting which patients with hip osteoarthritis are more likely to show disease progression is important for healthcare professionals. Therefore, the aim of this review was to assess which factors are predictive of progression in patients with hip osteoarthritis. A literature search was made up until 14 March 2019. Included were cohort and case-control studies evaluating the association between factors and progression (either clinical, radiological, or THR). Excluded were studies with a follow-up < 1 year or specific underlying pathologies of osteoarthritis. Risk of bias was assessed using the QUIPS tool. A best-evidence synthesis was conducted. We included 57 articles describing 154 different factors. Of these, a best-evidence synthesis was possible for 103 factors, separately for clinical and radiological progression, and progression to total hip replacement. We found strong evidence for more clinical progression in patients with comorbidity and more progression to total hip replacement for a higher Kellgren and Lawrence grade, superior or (supero) lateral femoral head migration, and subchondral sclerosis. Strong evidence for no association was found regarding clinical progression for gender, social support, pain medication, quality of life, and limited range of motion of internal rotation or external rotation. Also, strong evidence for no association was found regarding radiological progression for the markers CTX-I, COMP, NTX-I, PINP, and PIIINP and regarding progression to total hip replacement for body mass index. Strong evidence suggested that 4 factors were predictive of progression of hip osteoarthritis, whereas 12 factors were not predictive of progression. Evidence for most of the reported factors was either limited or conflicting. PROSPERO, CRD42015010757.


The Mental Health in Austrian Teenagers (MHAT)-Study: preliminary results from a pilot study.
No epidemiological data on prevalence rates of mental disorders based on a representative sample are available for Austrian adolescents up to now. However, the knowledge of psychiatric disorders, related risk and protective factors is of great significance for treatment and prevention. The purpose of the MHAT-Study (Mental Health in Austrian Teenagers), the first epidemiological study on mental health in Austria, is to obtain prevalence rates of mental disorders and to examine risk factors, protective factors and quality of life in a representative sample of adolescents aged 10-18. Aims of this pilot study were to evaluate the feasibility and acceptability of the screening instruments, pre-estimate the frequency of mental health problems and estimate possible non-responder bias. Twenty-one schools in eastern Austria were asked to participate. Data on mental health problems were derived from self-rating questionnaires containing standardized screening measures (Youth Self-Report, measuring emotional and behavioral problems and the SCOFF, indicating eating problems. Quality of life as well as related risk and protective factors were also obtained. Four hundred and eight adolescents of five schools were recruited. The prevalence of mental health problems was 18.9 % [CI 95 %: 14.9-22.7]. Moreover, emotional and behavioral problems were highly correlated with quality of life measures. A Non-Responder Analysis indicated that non-responders (16.7 %) differ from responders with regard of school related problems. The results demonstrate that mental health problems affect approximately one fifth of the adolescents. A Non-Responder Analysis suggests that the prevalence of behavioral and emotional problems is underestimated.


Doping control of testosterone and human chorionic gonadotrophin: a case study.
Doping control for testosterone and human Chorionic Gonadotrophin (hCG) requires special attention as a difference must be made between the endogenous and exogenous origin of both substances. The detection of exogenous testosterone is based on the ratio of testosterone- to epitestosterone-glucuronide (T/E) in urine. The problems with this ratio are discussed. For hCG analysis in urine the utilization of sandwich-type hCG specific assays instead of hCG/hCG beta competitive assays is recommended. A case study in which an athlete self-administered testosterone and hCG before a competition is described. The T/E ratio and hCG concentration in urine were followed during this period of self-administration. The results demonstrate the relevance of the T/E ratio and of the selected hCG assay. The ratio of testosterone to human Luteinizing Hormone (T/hLH) in serum also indicated the use of hormones. Although the athlete's urine was negative for exogenous testosterone directly after competition, he would have been found positive for hCG.


Effects of protonation state on a tyrosine-histidine bioinspired redox mediator.
The conversion of tyrosine to the corresponding tyrosyl radical in photosystem II (PSII) is an example of proton-coupled electron transfer. Although the tyrosine moiety (Tyr(Z)) is known to function as a redox mediator between the photo-oxidized primary donor (P680(•+)) and the Mn-containing oxygen-evolving complex, the protonation states involved in the course of the reaction remain an active area of investigation. Herein, we report on the optical, structural, and electrochemical properties of tyrosine-histidine constructs, which model the function of their naturally occurring counterparts in PSII. Electrochemical studies show that the phenoxyl/phenol couple of the model is chemically reversible and thermodynamically capable of water oxidation. Studies under acidic and basic conditions provide clear evidence that an ionizable proton controls the electrochemical potential of the tyrosine-histidine mimic and that an exogenous base or acid can be used to generate a low-potential or high-potential mediator, respectively. The phenoxyl/phenoxide couple associated with the low-potential mediator is thermodynamically incapable of water oxidation, whereas the relay associated with the high-potential mediator is thermodynamically incapable of reducing an attached photoexcited porphyrin. These studies provide insight regarding the mechanistic role of the tyrosine-histidine complex in water oxidation and strategies for making use of hydrogen bonds to affect the coupling between proton and electron transfer in artificial photosynthetic systems.


Factor analytic and reduced animal models for the investigation of additive genotype-by-environment interaction in outcrossing plant species with application to a Pinus radiata breeding programme.
Modelling additive genotype-by-environment interaction is best achieved with the use of factor analytic models. With numerous environments and for outcrossing plant species, computation is facilitated using reduced animal models. The development of efficient plant breeding strategies requires a knowledge of the magnitude and structure of genotype-by-environment interaction. This information can be obtained from appropriate linear mixed model analyses of phenotypic data from multi-environment trials. The use of factor analytic models for genotype-by-environment effects is known to provide a reliable, parsimonious and holistic approach for obtaining estimates of genetic correlations between all pairs of trials. When breeding for outcrossing species the focus is on estimating additive genetic correlations and effects which is achieved by including pedigree information in the analysis. The use of factor analytic models in this setting may be computationally prohibitive when the number of environments is moderate to large. In this paper, we present an approach that uses an approximate reduced animal model to overcome the computational issues associated with factor analytic models for additive genotype-by-environment effects. The approach is illustrated using a Pinus radiata breeding dataset involving 77 trials, located in environments across New Zealand and south eastern Australia, and with pedigree information on 315,581 trees. Using this approach we demonstrate the existence of substantial additive genotype-by-environment interaction for the trait of stem diameter measured at breast height. This finding has potentially significant implications for both breeding and deployment strategies. Although our approach has been developed for forest tree breeding programmes, it is directly applicable for other outcrossing plant species, including sugarcane, maize and numerous horticultural crops.


Snake bite in Chittagong Division, Bangladesh: a study of bitten patients who developed no signs of systemic envenoming.
The demographics, epidemiology, first aid, clinical management, treatment and outcome of snake bites causing no significant signs of systemic envenoming were documented in Chittagong Medical College Hospital, Bangladesh, between May 1999 and October 2002. Among 884 patients admitted, 350 were systemically envenomed and 534 were without signs of either systemic or significant local envenoming. The average age of patients with physical evidence of snake bite but no systemic envenoming was 26.4 years. Most had been bitten on their feet or hands. Ligatures had been applied proximal to the bite site in >95% of cases and the bite site had been incised in 13%. Patients were typically discharged at 24h. Those with clinical signs of systemic envenoming resembled the non-envenomed cases demographically and epidemiologically except that they arrived at hospital significantly later than non-envenomed patients, having spent longer with traditional healers. No non-envenomed patient was treated with antivenom and none went on to develop symptoms of systemic envenoming after discharge. The potential complications and confusing signs caused by ligatures and incision demand that all patients admitted with a history of snake bite be kept under observation for 24h after admission even if they have no signs of systemic envenoming.


The Parkin co-regulated gene product, PACRG, is an evolutionarily conserved axonemal protein that functions in outer-doublet microtubule morphogenesis.
Eukaryotic cilia and flagella are highly conserved structures composed of a canonical 9+2 microtubule axoneme. Comparative genomics of flagellated and non-flagellated eukaryotes provides one way to identify new putative flagellar proteins. We identified the Parkin co-regulated gene, or PACRG, from such a screen. Male mice deficient in PACRG are sterile, but its function has been little explored. The flagellated protozoan parasite Trypanosoma brucei possesses two homologues of PACRG. We performed RNA interference knockdown experiments of the two genes independently and both together. Simultaneous ablation of both proteins produced slow growth and paralysis of the flagellum with consequent effects on organelle segregation. Moreover, using transmission electron microscopy, structural defects were seen in the axoneme, with microtubule doublets missing from the canonical 9+2 formation. The occurrence of missing doublets increased toward the distal end of the flagellum and sequential loss of doublets was observed along individual axonemes. GFP fusion proteins of both PACRG homologues localised along the full length of the axoneme. Our results provide the first evidence for PACRG function within the axoneme, where we suggest that PACRG acts to maintain functional stability of the axonemal outer doublets of both motile and sensory cilia and flagella.


Trends in hepatocyte growth factor, insulin-like growth factor 1, thyroid-stimulating hormone, and leptin expression levels in uveal melanoma patient serum and tumor tissues: correlation to disease progression.
This exploratory study was carried out to determine the expression levels of hepatocyte growth factor (HGF), insulin-like growth factor 1, thyroid-stimulating hormone (TSH), and leptin in serum and tumor samples from patients with uveal melanoma and to investigate the potential association of these expression levels with disease progression and patient survival. Seventeen patients, including nine nonmetastatic and eight metastatic, were included in the study. Eighteen healthy individuals served as controls. The levels of these four proteins in serum and tissue samples were determined by enzyme-linked immunosorbent assays and immunohistochemical staining, respectively. Associations between protein levels and survival, disease progression, and other clinicopathological factors were analyzed statistically. Serum levels of HGF were significantly higher and TSH levels were lower in uveal melanoma patients than in healthy individuals, but the level of neither protein differed significantly between metastatic and nonmetastatic groups. Of the four proteins tested, only serum TSH was significantly associated with patient survival. No correlation was observed between the tissue and serum levels of each protein. The levels of HGF in serum may be markers of uveal melanoma development. The prognostic and predictive values of these potential markers need to be determined in a larger cohort.


Conformational rearrangements in the transmembrane domain of CNGA1 channels revealed by single-molecule force spectroscopy.
Cyclic nucleotide-gated (CNG) channels are activated by binding of cyclic nucleotides. Although structural studies have identified the channel pore and selectivity filter, conformation changes associated with gating remain poorly understood. Here we combine single-molecule force spectroscopy (SMFS) with mutagenesis, bioinformatics and electrophysiology to study conformational changes associated with gating. By expressing functional channels with SMFS fingerprints in Xenopus laevis oocytes, we were able to investigate gating of CNGA1 in a physiological-like membrane. Force spectra determined that the S4 transmembrane domain is mechanically coupled to S5 in the closed state, but S3 in the open state. We also show there are multiple pathways for the unfolding of the transmembrane domains, probably caused by a different degree of α-helix folding. This approach demonstrates that CNG transmembrane domains have dynamic structure and establishes SMFS as a tool for probing conformational change in ion channels.


Demographic, structural and genetic predictors of late cognitive decline after penetrating head injury.
We examined the relationship of pre-injury intelligence, demographic variables, lesion location, brain tissue volume loss and a number of genetic markers to long-term cognitive decline in a group of Vietnam veterans with predominantly penetrating head injury (PHI) suffered more than 30 years ago. Using linear and stepwise regression procedures, we found that those with PHI demonstrated a greater degree of cognitive decline overall during the years following recovery from injury compared with a control group of uninjured Vietnam veterans. This became increasingly significant later in life. We also found that pre-injury intelligence was the most consistent predictor of cognitive outcome across all phases of potential recovery and decline after such injuries. While laterality of lesion was not a factor, we did find some associations between atrophy and specific regions of tissue loss and long-term cognitive functioning. Finally, we found evidence for an association between level of cognitive decline following PHI and the possession of certain genetic markers that have been linked with brain injury and neurodegeneration. Thus exacerbated decline does occur in Vietnam veterans with PHI and it is apparently unrelated to dementia and is determined by multiple factors (most notably pre-injury intelligence).


How a Dove prism transforms the orbital angular momentum of a light beam.
It is generally assumed that a light beam with orbital angular momentum (OAM) per photon of lh, is transformed, when traversing a Dove prism, into a light beam with OAM per photon of -lh. In this paper, we show theoretically and experimentally that this OAM transformation rule does not apply for highly focused light beams. This result should be taken into account when designing classical and quantum algorithms that make use of Dove prims to manipulate the OAM of light.


[Physical activity behavior in the Swiss population: what do we know in 2010?].
Since 1999 Switzerland has physical activity recommendations for adults, since 2006 also for school-age children and adolescents. The best available estimates indicate that 32% of adults meet the recommendations for three endurance-type training sessions and another 9% the minimal recommendations of half an hour moderate intensity physical activity on most days of the week. 13% of adolescents seem to meet the minimal recommendations of one hour on every day of the week, 35% on five days per week. All approaches for physical activity promotion are very important, with the medical care settings having a particular role in reaching inactive individuals.


The influence of obesity on the outcome of treatment of lumbar disc herniation: analysis of the Spine Patient Outcomes Research Trial (SPORT).
Questions remain as to the effect that obesity has on patients managed for symptomatic lumbar disc herniation. The purpose of this study was to determine if obesity affects outcomes following the treatment of symptomatic lumbar disc herniation. An as-treated analysis was performed on patients enrolled in the Spine Patient Outcomes Research Trial for the treatment of lumbar disc herniation. A comparison was made between patients with a body mass index of <30 kg/m² (nonobese) (n = 854) and those with a body mass index of ≥30 kg/m² (obese) (n = 336). Baseline patient demographic and clinical characteristics were documented. Primary and secondary outcomes were measured at baseline and at regular follow-up time intervals up to four years. The difference in improvement from baseline between operative and nonoperative treatment was determined at each follow-up period for both groups. At the time of the four-year follow-up evaluation, improvements over baseline in primary outcome measures were significantly less for obese patients as compared with nonobese patients in both the operative treatment group (Short Form-36 physical function, 37.3 compared with 47.7 points [p < 0.001], Short Form-36 bodily pain, 44.2 compared with 50.0 points [p = 0.005], and Oswestry Disability Index, -33.7 compared with -40.1 points [p < 0.001]) and the nonoperative treatment group (Short Form-36 physical function, 23.1 compared with 32.0 points [p < 0.001] and Oswestry Disability Index, -21.4 compared with -26.1 points [p < 0.001]). The one exception was that the change from baseline in terms of the Short Form-36 bodily pain score was statistically similar for obese and nonobese patients in the nonoperative treatment group (30.9 compared with 33.4 points [p = 0.39]). At the time of the four-year follow-up evaluation, when compared with nonobese patients who had been managed operatively, obese patients who had been managed operatively had significantly less improvement in the Sciatica Bothersomeness Index and the Low Back Pain Bothersomeness Index, but had no significant difference in patient satisfaction or self-rated improvement. In the present study, 77.5% of obese patients and 86.9% of nonobese patients who had been managed operatively were working a full or part-time job. No significant differences were observed in the secondary outcome measures between obese and nonobese patients who had been managed nonoperatively. The benefit of surgery over nonoperative treatment was not affected by body mass index. Obese patients realized less clinical benefit from both operative and nonoperative treatment of lumbar disc herniation. Surgery provided similar benefit over nonoperative treatment in obese and nonobese patients.


Chronic leg ulcers in adult patients with rheumatological diseases - a 7-year retrospective review.
Chronic leg ulcers in patients with rheumatological diseases can cause significant morbidity. We performed a retrospective case review to describe the epidemiology, clinical features and outcome of chronic leg ulcers in this group of patients. Twenty-nine patients with underlying rheumatological conditions, such as, rheumatoid arthritis (15 patients), systemic lupus erythematosus (8 patients), overlap syndromes (3 patients), systemic sclerosis (1 patient) and ankylosing spondylitis (1 patient) were included. The ulcers were mostly located around the ankle (55·2%) and calves (37·9%). The predominant aetiology of the ulcers, in decreasing order of frequency, was venous disease, multifactorial, vasculitis or vasculopathy, infective, pyoderma gangrenosum, ischaemic microangiopathy and iatrogenic. Treatment modalities included aggressive wound bed preparation, compression therapy (17 patients), changes in immunosuppressive therapy (15 patients), hyperbaric oxygen therapy (4 patients) and cellular skin grafting (2 patients). Management of chronic leg ulcers in rheumatological patients is challenging and the importance of careful clinicopathological correlation and treatment of the underlying cause cannot be overemphasised.


Neural coding of formant-exaggerated speech in the infant brain.
Speech scientists have long proposed that formant exaggeration in infant-directed speech plays an important role in language acquisition. This event-related potential (ERP) study investigated neural coding of formant-exaggerated speech in 6-12-month-old infants. Two synthetic /i/ vowels were presented in alternating blocks to test the effects of formant exaggeration. ERP waveform analysis showed significantly enhanced N250 for formant exaggeration, which was more prominent in the right hemisphere than the left. Time-frequency analysis indicated increased neural synchronization for processing formant-exaggerated speech in the delta band at frontal-central-parietal electrode sites as well as in the theta band at frontal-central sites. Minimum norm estimates further revealed a bilateral temporal-parietal-frontal neural network in the infant brain sensitive to formant exaggeration. Collectively, these results provide the first evidence that formant expansion in infant-directed speech enhances neural activities for phonetic encoding and language learning.


Endocranial Development in the Coyote (Canis latrans) and Gray Wolf (Canis lupus): A Computed Tomographic Study.
The purpose of this study was to examine the pattern of postnatal brain growth in two wild canid species: the coyote (Canis latrans) and gray wolf (Canis lupus). Adult regional and total brain volume differences were also compared between the two species as well as within each species by sex. Three-dimensional virtual endocasts of endocranial airspace were created from computed tomography scans of 52 coyote skulls (28 female, 24 male; 1 day to 13.4 years) and 46 gray wolf skulls (25 female, 21 male; 1 day to 7.9 years). Age was known in coyotes or estimated from dentition patterns in wolves. The 95% asymptotic growth of the endocranium is completed by 21 weeks in male and 17.5 weeks in female coyotes and by 27 weeks in male and 18.5 weeks in female wolves. These ages are well before age at first reproduction (coyote - 40.4 weeks; wolf - 91.25 weeks). Skull growth as measured by centroid size lags behind endocranial growth but is also completed before sexual maturity. Intra- and interspecific comparisons of brain volumes in the adult wolves and coyotes revealed that relative anterior cerebrum (AC) volume was greater in males than females in both species. Relative brain size was greater in the coyote than in the wolf as was relative cerebrum volume. However, relative AC volume and relative cerebellum and brainstem volume was greater in the wolf than coyote. One explanation for the increased AC volume in males compared to females may be related to the role of social information processing. However, additional data are needed to determine the correspondence between regional volumes and functional differences either between or within these species. Nonetheless, these findings provide important baseline data for further studies on wild canid brain variations and development.


Dedifferentiated chondrosarcomas arising in preexisting osteochondromas.
Dedifferentiated chondrosarcomas that arise in osteochondromas are extremely rare lesions for which very little information on treatment and outcome is available in the literature. The purpose of the present study was to describe the specific clinical, radiographic, and histologic features of this lesion and to evaluate the oncologic outcome after different treatment strategies. We reviewed the files of the Rizzoli Institute between 1970 and 2002 and identified eighteen patients for whom adequate records and histologic images were available and in whom a high-grade sarcoma had been diagnosed at the same location as a preexisting osteochondroma. Radiographic studies, histologic slides, and clinical records were reviewed, the features of those studies were tabulated, and prognostic features and the results of treatment were identified. The patients included twelve men and six women with an average age of forty-six years (range, twenty-two to seventy-four years). Eight lesions occurred in patients with multiple osteochondromas, and ten occurred in patients with solitary lesions. The most common locations were the pelvis (six lesions) and the femur (five lesions). Symptoms included pain, swelling, and a growing mass; the average duration of symptoms was eighteen months. Radiographically, ten lesions appeared as a conventional secondary chondrosarcoma arising in an exostosis, whereas eight showed typical signs of dedifferentiation. Histologic evaluation of the cartilage component demonstrated thirteen grade-1 and two grade-2 chondrosarcomas. In three cases, no cartilage component was recognized. The dedifferentiated component was considered to be an osteosarcoma in nine cases (including six cases in which it was osteoblastic and three in which it was fibroblastic), a malignant fibrous histiocytoma in eight, and a fibrosarcoma in one. The dedifferentiated component represented an average of 59% (range, 20% to 100%) of the lesion. For the fifteen patients who were managed at the authors' institution, the two and five-year survival rates were 47% and 29%, respectively; the median survival time was fourteen months. Patients who were managed with a combination of surgery and chemotherapy had a better overall survival rate than did those who were managed with surgery alone (p = 0.03). Dedifferentiated chondrosarcoma arising in a preexisting osteochondroma is an extremely rare lesion with a poor prognosis. In the present small series, overall survival was better when wide surgical resection was combined with adjuvant chemotherapy.


Molecular analysis of the cumulus matrix: insights from mice with O-glycan-deficient oocytes.
During follicle development, oocytes secrete factors that influence the development of granulosa and cumulus cells (CCs). In response to oocyte and somatic cell signals, CCs produce extracellular matrix (ECM) molecules resulting in cumulus expansion, which is essential for ovulation, fertilisation, and is predictive of oocyte quality. The cumulus ECM is largely made up of hyaluronan (HA), TNF-stimulated gene-6 (TSG-6, also known as TNFAIP6), pentraxin-3 (PTX3), and the heavy chains (HCs) of serum-derived inter-α-inhibitor proteins. In contrast to other in vivo models where modified expansion impairs fertility, the cumulus mass of C1galt1 Mutants, which have oocyte-specific deletion of core 1-derived O-glycans, is modified without impairing fertility. In this report, we used C1galt1 Mutant (C1galt1(FF):ZP3Cre) and Control (C1galt1(FF)) mice to investigate how cumulus expansion is affected by oocyte-specific deletion of core 1-derived O-glycans without adversely affecting oocyte quality. Mutant cumulus-oocyte complexes (COCs) are smaller than Controls, with fewer CCs. Interestingly, the CCs in Mutant mice are functionally normal as each cell produced normal levels of the ECM molecules HA, TSG-6, and PTX3. However, HC levels were elevated in Mutant COCs. These data reveal that oocyte glycoproteins carrying core 1-derived O-glycans have a regulatory role in COC development. In addition, our study of Controls indicates that a functional COC can form provided all essential components are present above a minimum threshold level, and thus some variation in ECM composition does not adversely affect oocyte development, ovulation or fertilisation. These data have important implications for IVF and the use of cumulus expansion as a criterion for oocyte assessment.


Ventral striatum and amygdala activity as convergence sites for early adversity and conduct disorder.
Childhood family adversity (CFA) increases the risk for conduct disorder (CD) and has been associated with alterations in regions of affective processing like ventral striatum (VS) and amygdala. However, no study so far has demonstrated neural converging effects of CFA and CD in the same sample. At age 25 years, functional MRI data during two affective tasks, i.e. a reward (N = 171) and a face-matching paradigm (N = 181) and anatomical scans (N = 181) were acquired in right-handed currently healthy participants of an epidemiological study followed since birth. CFA during childhood was determined using a standardized parent interview. Disruptive behaviors and CD diagnoses during childhood and adolescence were obtained by diagnostic interview (2-19 years), temperamental reward dependence was assessed by questionnaire (15 and 19 years).CFA predicted increased CD and amygdala volume. Both exposure to CFA and CD were associated with a decreased VS response during reward anticipation and blunted amygdala activity during face-matching. CD mediated the effect of CFA on brain activity. Temperamental reward dependence was negatively correlated with CFA and CD and positively with VS activity. These findings underline the detrimental effects of CFA on the offspring's affective processing and support the importance of early postnatal intervention programs aiming to reduce childhood adversity factors.


The Nucleotide-Binding Sites of SUR1: A Mechanistic Model.
ATP-sensitive potassium (KATP) channels comprise four pore-forming Kir6.2 subunits and four modulatory sulfonylurea receptor (SUR) subunits. The latter belong to the ATP-binding cassette family of transporters. KATP channels are inhibited by ATP (or ADP) binding to Kir6.2 and activated by Mg-nucleotide interactions with SUR. This dual regulation enables the KATP channel to couple the metabolic state of a cell to its electrical excitability and is crucial for the KATP channel's role in regulating insulin secretion, cardiac and neuronal excitability, and vascular tone. Here, we review the regulation of the KATP channel by adenine nucleotides and present an equilibrium allosteric model for nucleotide activation and inhibition. The model can account for many experimental observations in the literature and provides testable predictions for future experiments.


Autoantibody profiles reveal ubiquilin 1 as a humoral immune response target in lung adenocarcinoma.
There is considerable evidence that the presence of cancer can elicit a humoral immune response to specific proteins in the host, and these resulting autoantibodies may have potential as noninvasive biomarkers. To characterize the autoantibody repertoire present in the sera of patients with lung adenocarcinoma, we developed a high-density peptide microarray derived from biopanning a lung cancer phage display library. Using a 2,304-element microarray, we interrogated a total of 250 sera from Michigan lung cancer patients and noncancer controls to develop an "autoantibody profile" of lung adenocarcinoma. A set of 22 discriminating peptides derived from a training set of 125 serum samples from lung adenocarcinoma patients and control subjects was found to predict cancer status with 85% sensitivity and 86% specificity in an independent test set of 125 sera. Sequencing of the immunoreactive phage-peptide clones identified candidate humoral immune response targets in lung adenocarcinoma, including ubiquilin 1, a protein that regulates the degradation of several ubiquitin-dependent proteasome substrates. An independent validation set of 122 serum samples from Pittsburgh was examined using two overlapping clones of ubiquilin 1 that showed 0.79 and 0.74 of the area under the receiver operating characteristics curve, respectively. Significantly increased levels of both ubiquilin 1 mRNA and protein, as well as reduced levels of the phosphorylated form of this protein, were detected in lung tumors. Immunofluorescence using anti-ubiquilin 1 antibodies confirmed intracellular expression within tumors cells. These studies indicate that autoantibody profiles, as well as individual candidates, may be useful for the noninvasive detection of lung adenocarcinoma.


Reduction strategies in animal research: a review of scientific approaches at the intra-experimental, supra-experimental and extra-experimental levels.
When discussing animal use and considering alternatives to animals in biomedical research and testing, the number of animals required gets to the root of the matter on ethics and justification. In this paper, some reduction strategies are reviewed. Many articles and reports on reduction of animal use focus mostly on the experimental level, but other approaches are also possible. Reduction at the intraexperimental level probably offers the greatest scope for reduction, as the design and statistical analysis of individual experiments can often be improved. Supra-experimental reduction aims to reduce the number of animals by a change in the setting in which a series of experiments take place--for example, by improved education and training, reduction of breeding surpluses, critical analysis of test specifications, and re-use of animals. At the extra-experimental level, reduction is a spin-off of other developments, rather than the direct goal. Through improved research or production strategies, aimed at better quality, consistency and safety, reduction in the number of animals used can be substantial. A revised definition of reduction is proposed, which does not include the level of information needed, as in some cases reduction in the number of animals resulting in less information or data, is still acceptable.


New "light" for one-world approach toward safe and effective control of animal diseases and insect vectors from leishmaniac perspectives.
Light is known to excite photosensitizers (PS) to produce cytotoxic reactive oxygen species (ROS) in the presence of oxygen. This modality is attractive for designing control measures against animal diseases and pests. Many PS have a proven safety record. Also, the ROS cytotoxicity selects no resistant mutants, unlike other drugs and pesticides. Photodynamic therapy (PDT) refers to the use of PS as light activable tumoricides, microbicides and pesticides in medicine and agriculture.Here we describe "photodynamic vaccination" (PDV) that uses PDT-inactivation of parasites, i.e. Leishmania as whole-cell vaccines against leishmaniasis, and as a universal carrier to deliver transgenic add-on vaccines against other infectious and malignant diseases. The efficacy of Leishmania for vaccine delivery makes use of their inherent attributes to parasitize antigen (vaccine)-presenting cells. Inactivation of Leishmania by PDT provides safety for their use. This is accomplished in two different ways: (i) chemical engineering of PS to enhance their uptake, e.g. Si-phthalocyanines; and (ii) transgenic approach to render Leishmania inducible for porphyrinogenesis. Three different schemes of Leishmania-based PDV are presented diagrammatically to depict the cellular events resulting in cell-mediated immunity, as seen experimentally against leishmaniasis and Leishmania-delivered antigen in vitro and in vivo. Safety versus efficacy evaluations are under way for PDT-inactivated Leishmania, including those further processed to facilitate their storage and transport. Leishmania transfected to express cancer and viral vaccine candidates are being prepared accordingly for experimental trials.We have begun to examine PS-mediated photodynamic insecticides (PDI). Mosquito cells take up rose bengal/cyanosine, rendering them light-sensitive to undergo disintegration in vitro, thereby providing a cellular basis for the larvicidal activity seen by the same treatments. Ineffectiveness of phthalocyanines and porphyrins for PDI underscores its requirement for different PS. Differential uptake of PS by insect versus other cells to account for this difference is under study.The ongoing work is patterned after the one-world approach by enlisting the participation of experts in medicinal chemistry, cell/molecular biology, immunology, parasitology, entomology, cancer research, tropical medicine and veterinary medicine. The availability of multidisciplinary expertise is indispensable for implementation of the necessary studies to move the project toward product development.


"Choking game" awareness and participation among 8th graders--Oregon, 2008.
The "choking game" is an activity in which persons strangulate themselves to achieve euphoria through brief hypoxia. It is differentiated from autoerotic asphyxiation. The activity can cause long-term disability and death among youths. In 2008, CDC reported 82 deaths attributed to the choking game and other strangulation activities during the period 1995-2007; most victims were adolescent males aged 11-16 years. To assess the awareness and prevalence of this behavior among 8th graders in Oregon, the Oregon Public Health Division added a question to the 2008 Oregon Healthy Teens survey concerning familiarity with and participation in this activity. This report describes the results of that survey, which indicated that 36.2% of 8th-grade respondents had heard of the choking game, 30.4% had heard of someone participating, and 5.7% had participated themselves. Youths in rural areas were significantly more likely (6.7%) to have participated than youths in urban areas (4.9%). Choking game participation was higher among 8th graders who reported mental health risk factors (4.0%), substance use (7.9%), or both (15.8%), compared with those who reported neither (1.7%). Public health surveillance of these strangulation activities among youths should be expanded to better quantify the risks and understand the motives and circumstances surrounding participation. Parents, educators, counselors, and others who work with youths should be aware of strangulation activities and their serious health effects; they should watch for signs of participation in strangulation activities, especially among youths with suspected substance use or mental health risk factors.


Biliverdin Reductase A (BVRA) Mediates Macrophage Expression of Interleukin-10 in Injured Kidney.
Biliverdin reductase A is an enzyme, with serine/threonine/tyrosine kinase activation, converting biliverdin (BV) to bilirubin (BR) in heme degradation pathway. It has been reported to have anti-inflammatory and antioxidant effect in monocytes and human glioblastoma. However, the function of BVRA in polarized macrophage was unknown. This study aimed to investigate the effect of BVRA on macrophage activation and polarization in injured renal microenvironment. Classically activated macrophages (M1macrophages) and alternative activation of macrophages (M2 macrophages) polarization of murine bone marrow derived macrophage was induced by GM-CSF and M-CSF. M1 polarization was associated with a significant down-regulation of BVRA and Interleukin-10 (IL-10), and increased secretion of TNF-α. We also found IL-10 expression was increased in BVRA over-expressed macrophages, while it decreased in BVRA knockdown macrophages. In contrast, BVRA over-expressed or knockdown macrophages had no effect on TNF-α expression level, indicating BVRA mediated IL-10 expression in macrophages. Furthermore, we observed in macrophages infected with recombinant adenoviruses BVRA gene, which BVRA over-expressed enhanced both INOS and ARG-1 mRNA expression, resulting in a specific macrophage phenotype. Through in vivo study, we found BVRA positive macrophages largely existed in mice renal ischemia perfusion injury. With the treatment of the regular cytokines GM-CSF, M-CSF or LPS, excreted in the injured renal microenvironment, IL-10 secretion was significantly increased in BVRA over-expressed macrophages. In conclusion, the BVRA positive macrophage is a source of anti-inflammatory cytokine IL-10 in injured kidney, which may provide a potential target for treatment of kidney disease.