Associations of SNPs in ADIPOQ and subclinical cardiovascular disease in the multi-ethnic study of atherosclerosis (MESA).
Circulating adiponectin is associated with both clinical and subclinical cardiovascular disease (CVD). Variants of the adiponectin gene (ADIPOQ) are associated with clinical CVD, but little is known about associations with subclinical CVD. We studied the association of 11 ADIPOQ single-nucleotide polymorphisms (SNPs) with common and internal carotid intima media thickness (cIMT), presence of coronary artery calcification (CAC), and CAC scores (in those with CAC) in 2,847 participants in the Multi-Ethnic Study of Atherosclerosis (MESA). Participants were white (n = 712), African American (n = 712), Chinese (n = 718), and Hispanic (n = 705). All models were adjusted for age, sex, and field site, and stratified by race/ethnic group. African Americans with genotypes AG/GG of rs2241767 had 36% greater (95% confidence interval (CI; 16%, 59%), P = 0.0001) CAC prevalence; they also had a larger common cIMT (P = 0.0043). Also in African Americans, genotypes AG/AA of rs1063537 were associated with a 35% (95% CI (14%, 59%), P = 0.0005) greater CAC prevalence. Hispanics with the AA genotype of rs11711353 had a 37% (95% CI (14%, 66%), P = 0.0011), greater CAC prevalence compared to those with the GG genotype. Additional adjustment for ancestry in African-American and Hispanic participants did not change the results. No single SNP was associated with subclinical CVD phenotypes in Chinese or white participants. There appears to be an association between ADIPOQ SNPs and subclinical CVD in African Americans and Hispanics. Replication as well as assessment of other ADIPOQ SNPs is warranted.


TLE1 modifies the effects of NOD2 in the pathogenesis of Crohn's disease.
The mechanisms by which specific mutations in NOD2/CARD15 increase the risk for Crohn's disease (CD) are unclear. We identified proteins that interact with NOD2 and investigated them by expression, genetic, and functional analyses. By using a yeast 2-hybrid screen of an intestinal epithelial library, we identified proteins that interact with NOD2 and confirmed the interactions in mammalian cells using co-immunoprecipitation. We used microarray analysis to analyze gene expression patterns in 302 intestinal biopsy samples (129 from patients with ulcerative colitis [UC], 106 with CD, and 67 controls). Eighty single-nucleotide polymorphisms within the genes that encoded 6 interacting proteins were genotyped in a discovery cohort (869 cases of inflammatory bowel disease [IBD], 885 controls) and a replication cohort (504 patients with IBD, 713 controls). We investigated interaction between transducin-like enhancer of split 1 (TLE1) and NOD2 in HEK293 cells. We identified 6 NOD2-interacting proteins (TLE1, UDP-N-acetyl-alpha-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2 [GALNT2], HIV-1 Tat interactive protein [HTATIP], Vimentin, fission 1 (mitochondrial outer membrane) homolog [FIS1], and protein phosphatase 2, regulatory subunit B', epsilon isoform [PPP2R5E]). Of these, expression of GALNT2 (CD, P = .004) and vimentin (CD, P = .006; UC, P = .0025) was altered in patients with IBD compared with controls. Single-nucleotide polymorphisms within TLE1 were associated with susceptibility to CD, specifically with ileal disease (rs6559629, P = 3.1 × 10⁻⁵; odds ratio, 1.45). The TLE1 risk allele is required for susceptibility to CD in carriers of NOD2 mutations. In cells, TLE1 and NOD2 co-localized around the nuclear membrane and TLE1 inhibited activation of nuclear factor-κB by NOD2. Epistatic and biological interactions between TLE1 and NOD2 are involved in IBD pathogenesis. NOD2 might be involved in a series of pathways such as epigenetic regulation of expression (via TLE1 and HTATIP), biosynthesis of mucin (via GALNT2), apoptosis (via PPP2R5E and FIS1), and integrity of the intracellular cytoskeleton (vimentin).


Effects of self-administered alcohol concentration on the frequency and persistence of rats' attending to alcohol cues.
Observing responses produce contact with stimuli that are to be discriminated and have been considered an animal model of attending. In the observing-response procedure, alternating periods of drug availability versus extinction for one response are not signaled, but a second response (i.e. the observing response) produces stimuli signaling whether drug is available or not. This experiment examined the effects of the concentration of self-administered alcohol and increases in observing-response requirement on rats' observing alcohol stimuli. In addition, the effects of alcohol concentration on the persistence of observing were examined when alcohol was no longer available. Results showed that observing tracked bitonic changes in the number of alcohol deliveries rather than monotonic increases in total alcohol consumption resulting from increases in alcohol concentration. Increasing the observing-response requirement decreased the number of stimulus presentations earned. The resultant decreases in time spent in the presence of the alcohol stimulus were associated with decreases in alcohol consumption. During extinction of alcohol responding, observing was more persistent when it produced a stimulus previously associated with a higher alcohol concentration. Finally, responding for alcohol was more resistant to extinction in the presence of an observing-response-produced alcohol stimulus than in its absence, but did not depend on alcohol concentration. These results suggest that increases in the difficulty of obtaining access to alcohol cues can decrease alcohol consumption by reducing contact with those cues. In addition, if observing behavior in the present procedure is analogous to attending to alcohol cues, the results suggest that attending to alcohol cues is more persistent with cues previously associated with higher doses, and that the persistence of attending to alcohol cues and their impact on drinking may be dissociable.


[Augmentation with atypical antipsychotics for the treatment of patients with a therapy-resistant depression: a review].
A considerable number of depressed patients are showing resistance to current drug treatment strategies. In such cases it is becoming increasingly common in clinical practice to augment an antidepressant with an atypical antipsychotic (aap). purpose To provide an overview of the scientific evidence for this new strategy, to explain the possible mechanisms of action and to assess the place that augmentation with an aap occupies in the treatment of therapy-resistant non-psychotic depression. Various search terms were used to locate relevant articles in Pubmed; these articles were examined for relevant references. results Only 6 randomised controlled trials were found, therefore 7 case reports and 10 open-label studies were included. There seems to be some evidence that augmentation with an aap, particularly olanzapine, is effective. One of the main advantages of this strategy is the fast response, namely within a few weeks or even within a week. In view of the lack of scientific support for augmentation with aaps, this strategy is advisable when other augmentation strategies have proved unworkable or are contraindicated. It must be exercised with caution because the combination therapy can have negative effects on the patient's glucose and lipid metabolism. Augmentation with aaps in treatment resistant depression may be a potential useful treatment strategy but its scientific evidence is insufficient to warrant inclusion within current guidelines.


Functional localization of neurotransmitter receptors and synaptic inputs to mature neurons of the medial superior olive.
Neurons of the medial superior olive (MSO) code for the azimuthal location of low-frequency sound sources via a binaural coincidence detection system operating on microsecond time scales. These neurons are morphologically simple and stereotyped, and anatomical studies have indicated a functional segregation of excitatory and inhibitory inputs between cellular compartments. It is thought that this morphological arrangement holds important implications for the computational task of these cells. To date, however, there has been no functional investigation into synaptic input sites or functional receptor distributions on mature neurons of the MSO. Here, functional neurotransmitter receptor maps for amino-3-hydroxyl-5-methyl-4-isoxazole propionate (AMPA), N-methyl-D-aspartate (NMDA), glycine (Gly), and ionotropic γ-aminobutyric acid (GABA(A)) receptors (Rs) were compared and complemented by their corresponding synaptic input map. We find in MSO neurons from postnatal day 20-35 gerbils that AMPARs and their excitatory inputs target the soma and dendrites. Functional GlyRs and their inhibitory inputs are predominantly refined to the somata, although a pool of functional GlyRs is present extrasynaptically on MSO dendrites. GABA(A)R responses are present throughout the cell but lack direct synaptic contact indicating an involvement in volume transmission. NMDARs are present both synaptically and extrasynaptically with an overall distribution similar to GlyRs. Interestingly, even at physiological temperatures these functional NMDARs can be potentiated by synaptically released Gly. The functional receptor and synaptic input maps produced here led to the identification of a cross talk between transmitter systems and raises the possibility that extrasynaptic receptors could be modulating leak conductances as a homeostatic mechanism.


Benign breast tissue composition in breast cancer patients: association with risk factors, clinical variables, and gene expression.
Breast tissue composition (epithelium, non-fatty stroma, and adipose) changes qualitatively and quantitatively throughout the lifespan, and may mediate relationships between risk factors and breast cancer initiation. We sought to identify relationships between tissue composition, risk factors, tumor characteristics, and gene expression. Participants were 146 patients from the Polish Breast Cancer Study, with data on risk factor and clinicopathological characteristics. Benign breast tissue composition was evaluated using digital image analysis of histologic sections. Whole-genome microarrays were performed on the same tissue blocks. Mean epithelial, non-fatty stromal, and adipose proportions were 8.4% (SD = 4.9%), 27.7% (SD = 24.0%), and 64.0% (SD = 24.0%), respectively. Among women <50 years old, stroma proportion decreased and adipose proportion increased with age, with approximately 2% difference per year (P < 0.01). The variation in epithelial proportion with age was modest (0.1% per year). Higher epithelial proportion was associated with obesity (7.6% in nonobese vs. 10.1% in obese; P = 0.02) and with poorly differentiated tumors (7.8% in well/moderate vs. 9.9% in poor; P = 0.05). Gene expression signatures associated with epithelial and stromal proportion were identified and validated. Stroma-associated genes were in metabolism and stem cell maintenance pathways, whereas epithelial genes were enriched for cytokine and immune response pathways. Breast tissue composition was associated with age, body mass index, and tumor grade, with consequences for breast gene expression. Breast tissue morphologic factors may influence breast cancer etiology. Composition and gene expression may act as biomarkers of breast cancer risk and progression.


Antiviral resistance of biologic HIV-2 clones obtained from individuals on nucleoside reverse transcriptase inhibitor therapy.
To study phenotypic and genotypic resistance of HIV-2 against nucleoside reverse transcriptase inhibitors (NRTI). Biologic HIV-2 clones were generated from 3 patients before and after initiation of antiretroviral therapy with zidovudine (AZT) in patient RH2-7, AZT and didanosine (ddI) in patient PH2-1, and after addition of lamivudine (3TC) to AZT monotherapy in patient RH2-5. The sensitivity to NRTI of the virus clones, as defined by the 50% inhibitory concentration (IC(50)), was determined in vitro. The predicted amino acid sequences of the reverse transcriptase proteins from these clones were determined. Comparing the sensitivity of the biologic HIV-2 clones obtained after start of therapy with those from antiviral naive patients, resistance had developed to AZT (patients RH2-7 and RH2-5) and 3TC (patient PH2-1 and RH2-5). No resistance to AZT was observed in the biologic clone from PH2-1 obtained after start of therapy. The resistant clones from RH2-5 and PH2-1, but not RH2-7, contained amino acid mutations at positions where HIV-1 has been shown to mutate after AZT and 3TC treatment. Phenotypic resistance of HIV-2 to nucleoside analogues, which developed in HIV-2-infected patients treated with NRTIs, was associated with genotypic changes. Some of the mutations at amino acid positions in the HIV-2 reverse transcriptase gene corresponded with those involved in HIV-1 resistance, although no conventional mutations associated with resistance to AZT were observed.


Inferior colliculus lesions impair eyeblink conditioning in rats.
The neural plasticity necessary for acquisition and retention of eyeblink conditioning has been localized to the cerebellum. However, the sources of sensory input to the cerebellum that are necessary for establishing learning-related plasticity have not been identified completely. The inferior colliculus may be a source of sensory input to the cerebellum through its projection to the medial auditory thalamus. The medial auditory thalamus is necessary for eyeblink conditioning in rats and projects to the lateral pontine nuclei, which then project to the cerebellar nuclei and cortex. The current experiment examined the role of the inferior colliculus in auditory eyeblink conditioning. Rats were given bilateral or unilateral (contralateral to the conditioned eye) lesions of the inferior colliculus prior to 10 d of delay eyeblink conditioning with a tone CS. Rats with bilateral or unilateral lesions showed equivalently impaired acquisition. The extent of damage to the contralateral inferior colliculus correlated with several measures of conditioning. The findings indicate that the contralateral inferior colliculus provides auditory input to the cerebellum that is necessary for eyeblink conditioning.


Identification of fast and slow ventilatory responses to carbon dioxide under hypoxic and hyperoxic conditions in humans.
1. Under conditions of both euoxia and hypoxia, it is generally accepted that the ventilatory response to CO2 has both rapid (peripheral chemoreflex) and slow (central chemoreflex) components. However, under conditions of hyperoxia, it is unclear in humans whether the fast component is completely abolished or merely attenuated in magnitude. 2. The present study develops a technique to determine whether or not a two-compartment model fits the ventilatory response to CO2 significantly better than a one-compartment model. Data were collected under both hypoxic (end-tidal PO2 = 50 Torr) conditions, when two components would be expected, and under hyperoxic (end-tidal PO2 = 200 Torr) conditions, when the presence of the fast compartment is under question. 3. Ten subjects were recruited, of whom nine completed the study. The end-tidal PCO2 of each subject was varied according to a multi-frequency binary sequence that involved 13 steps into and 13 steps out of hypercapnia lasting altogether 1408 s. 4. In four out of nine subjects in hypoxia, and six out of nine subjects in hyperoxia, the two-compartment model fitted the data significantly better than the one-compartment model (F ratio test on residuals). This improvement in fit was significant for the pooled data in both hypoxia (P < 0.05) and hyperoxia (P < 0.005). Mean ventilatory sensitivities for the central chemoreflex were (mean +/- s.e.m.) 1. 69 +/- 0.39 l min-1 Torr-1 in hypoxia and 2.00 +/- 0.32 l min-1 Torr-1 in hyperoxia. Mean ventilatory sensitivities for the peripheral chemoreflex were 2.42 +/- 0.36 l min-1 Torr-1 in hypoxia and 0.75 +/- 0.16 l min-1 Torr-1 in hyperoxia. 5. It is concluded that the rapid and slow components of the ventilatory response to CO2 can be separately identified, and that a rapid component persists under conditions of hyperoxia.


Planar Asymmetries in the C. elegans Embryo Emerge by Differential Retention of aPARs at Cell-Cell Contacts.
Formation of the anteroposterior and dorsoventral body axis in Caenorhabditis elegans depends on cortical flows and advection of polarity determinants. The role of this patterning mechanism in tissue polarization after formation of cell-cell contacts is not fully understood. Here, we demonstrate that planar asymmetries are established during left-right symmetry breaking: Centripetal cortical flows asymmetrically and differentially advect anterior polarity determinants (aPARs) from contacts to the medial cortex, resulting in their unmixing from apical myosin. Contact localization and advection of PAR-6 requires balanced CDC-42 activation, while asymmetric retention and advection of PAR-3 can occur independently of PAR-6. Concurrent asymmetric retention of PAR-3, E-cadherin/HMR-1 and opposing retention of antagonistic CDC-42 and Wnt pathway components leads to planar asymmetries. The most obvious mark of planar asymmetry, retention of PAR-3 at a single cell-cell contact, is required for proper cytokinetic cell intercalation. Hence, our data uncover how planar polarity is established in a system without the canonical planar cell polarity pathway through planar asymmetric retention of aPARs.


Anterior positional fault of the fibula after sub-acute lateral ankle sprains.
Recent evidence has suggested a positional fault of the fibula exists in chronically unstable ankles. However, there has been little research examining positional faults after a sub-acute lateral ankle sprain (LAS). Our purpose was to measure the position of the distal fibula in relation to the distal tibia in subjects with sub-acute LASs and to determine if there is a relationship between the amount of swelling and fibular position. Eleven subjects with a unilateral sub-acute LAS and 11 healthy controls participated. The Wilcoxon signed rank test revealed a significant (p=0.008) difference within the ankles of the injured group. The sprained ankles had a mean fibular position of 14.2+3.4 mm and were positioned significantly more anteriorly than the contralateral uninjured ankles (17.0+3.2 mm). The Mann-Whitney test revealed a significant difference (p=0.045) between the sprained ankle and the side-matched limbs of the controls (16.8+2.3) Pearson product moment correlations revealed a strong positive correlation between fibular position and swelling (r=0.793, p=0.004). Those ankles with more swelling had the most anteriorly positioned fibulae. The fibulae in sub-acutely sprained ankles appear to be positioned more anteriorly compared to the contralateral ankles. This positional fault may be maintained acutely by swelling.


Subcallosal cingulate deep brain stimulation for treatment-refractory anorexia nervosa: a phase 1 pilot trial.
Anorexia nervosa is characterised by a chronic course that is refractory to treatment in many patients and has one of the highest mortality rates of any psychiatric disorder. Deep brain stimulation (DBS) has been applied to circuit-based neuropsychiatric diseases, such as Parkinson's disease and major depression, with promising results. We aimed to assess the safety of DBS to modulate the activity of limbic circuits and to examine how this might affect the clinical features of anorexia nervosa. We did a phase 1, prospective trial of subcallosal cingulate DBS in six patients with chronic, severe, and treatment-refractory anorexia nervosa. Eligible patients were aged 20-60 years, had been diagnosed with restricting or binge-purging anorexia nervosa, and showed evidence of chronicity or treatment resistance. Patients underwent medical optimisation preoperatively and had baseline body-mass index (BMI), psychometric, and neuroimaging investigations, followed by implantation of electrodes and pulse generators for continuous delivery of electrical stimulation. Patients were followed up for 9 months after DBS activation, and the primary outcome of adverse events associated with surgery or stimulation was monitored at every follow-up visit. Repeat psychometric assessments, BMI measurements, and neuroimaging investigations were also done at various intervals. This trial is registered with ClinicalTrials.gov, number NCT01476540. DBS was associated with several adverse events, only one of which (seizure during programming, roughly 2 weeks after surgery) was serious. Other related adverse events were panic attack during surgery, nausea, air embolus, and pain. After 9 months, three of the six patients had achieved and maintained a BMI greater than their historical baselines. DBS was associated with improvements in mood, anxiety, affective regulation, and anorexia nervosa-related obsessions and compulsions in four patients and with improvements in quality of life in three patients after 6 months of stimulation. These clinical benefits were accompanied by changes in cerebral glucose metabolism (seen in a comparison of composite PET scans at baseline and 6 months) that were consistent with a reversal of the abnormalities seen in the anterior cingulate, insula, and parietal lobe in the disorder. Subcallosal cingulate DBS seems to be generally safe in this sample of patients with chronic and treatment-refractory anorexia nervosa. Klarman Family Foundation Grants Program in Eating Disorders Research and Canadian Institutes of Health Research.


A multicentre parallel-group randomised trial assessing multiparametric MRI characterisation and image-guided biopsy of prostate in men suspected of having prostate cancer: MULTIPROS study protocol.
There is growing evidence suggesting that multiparametric magnetic resonance imaging (mpMRI) is a marker for prostate cancer (PCa) aggressiveness and could be used to plan treatment. Improving early detection of clinically significant PCa with pre-biopsy mpMRI would very likely have advantages including optimising the diagnosis and treatment of diseases and diminishing patient anxiety. This is a prospective multicentre study of pre-biopsy mpMRI diagnostic test accuracy with subgroup randomisation at a 1:1 ratio with respect to transrectal ultrasound (TRUS) and MRI/US fusion-guided biopsy or TRUS-only biopsy. It is designed as a single-gate study with a single set of inclusion criteria. The total duration of the recruitment phase was 48 months; however, this has now been extended to 66 months. A sample size of 600 participants is required. The primary objective is to determine whether mpMRI can improve PCa detection and characterisation. The key secondary objective is to determine whether MRI/US fusion-guided biopsy can reduce the number of false-negative biopsies. Ethical approval was obtained from the East of Scotland Research Ethics Committee 1 (14/ES/1070) on 20 November 2014. The results of this study will be used for publication and presentation in national and international journals and at scientific conferences. ClinicalTrials.gov, NCT02745496. Retrospectively registered on 20 April 2016.


Identifying pediatric community-acquired pneumonia hospitalizations: Accuracy of administrative billing codes.
Community-acquired pneumonia (CAP) remains one of the most common indications for pediatric hospitalization in the United States, and it is frequently the focus of research and quality studies. Use of administrative data is increasingly common for these purposes, although proper validation is required to ensure valid study conclusions. To validate administrative billing data for hospitalizations owing to childhood CAP. Case-control study of 4 tertiary care, freestanding children's hospitals in the United States. A total of 998 medical records of a 25% random sample of 3646 children discharged in 2010 with at least 1 International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) code representing possible pneumonia were reviewed. Discharges (matched on date of admission) without a pneumonia-related discharge code were also examined to identify potential missed pneumonia cases. Two reference standards, based on provider diagnosis alone (provider confirmed) or in combination with consistent clinical and radiographic evidence of pneumonia (definite), were used to identify CAP. Twelve ICD-9-CM–based coding strategies, each using a combination of primary or secondary codes representing pneumonia or pneumonia-related complications. Six algorithms excluded children with complex chronic conditions. Sensitivity, specificity, and negative and positive predictive values (NPV and PPV, respectively) of the 12 identification strategies. For provider-confirmed CAP (n = 680), sensitivity ranged from 60.7% to 99.7%; specificity, 75.7% to 96.4%; PPV, 67.9% to 89.6%; and NPV, 82.6% to 99.8%. For definite CAP (n = 547), sensitivity ranged from 65.6% to 99.6%; specificity, 68.7% to 93.0%; PPV, 54.6% to 77.9%; and NPV, 87.8% to 99.8%. Unrestricted use of the pneumonia-related codes was inaccurate, although several strategies improved specificity to more than 90% with a variable effect on sensitivity. Excluding children with complex chronic conditions demonstrated the most favorable performance characteristics. Performance of the algorithms was similar across institutions. Administrative data are valuable for studying pediatric CAP hospitalizations. The strategies presented here will aid in the accurate identification of relevant and comparable patient populations for research and performance improvement studies.


Intestinal degradation of dietary fibre in green beans--effects of microwave treatments.
Intestinal degradation of dietary fibre in blanched and microwaved green beans was studied by using a rat experimental model. Content and composition of dietary fibre as well as molecular weight distribution of water-soluble polysaccharides (WSP) were analysed. There was a solubilization and a shift towards lower Mw of mainly uronic acid-containing polysaccharides with repeated microwave treatment in the raw material. Thus, the apparent Mw of water-soluble polysaccharides decreased from approximately 1,550,000 to approximately 300,000. After the beans had been digested the Mw of the WSP was significantly reduced, to approximately 100,000, and the differences in Mw seen between various processed raw materials had been evened out. After fermentation the Mw of the WSP decreased further approximately 10 times. Fibre fermentability was high (approximately 90%) and similar for the various processed beans.


Biomechanical strain as a trigger for pore formation in Schlemm's canal endothelial cells.
The bulk of aqueous humor passing through the conventional outflow pathway must cross the inner wall endothelium of Schlemm's canal (SC), likely through micron-sized transendothelial pores. SC pore density is reduced in glaucoma, possibly contributing to obstructed aqueous humor outflow and elevated intraocular pressure (IOP). Little is known about the mechanisms of pore formation; however, pores are often observed near dome-like cellular outpouchings known as giant vacuoles (GVs) where significant biomechanical strain acts on SC cells. We hypothesize that biomechanical strain triggers pore formation in SC cells. To test this hypothesis, primary human SC cells were isolated from three non-glaucomatous donors (aged 34, 44 and 68), and seeded on collagen-coated elastic membranes held within a membrane stretching device. Membranes were then exposed to 0%, 10% or 20% equibiaxial strain, and the cells were aldehyde-fixed 5 min after the onset of strain. Each membrane contained 3-4 separate monolayers of SC cells as replicates (N = 34 total monolayers), and pores were assessed by scanning electron microscopy in 12 randomly selected regions (∼65,000 μm(2) per monolayer). Pores were identified and counted by four independent masked observers. Pore density increased with strain in all three cell lines (p < 0.010), increasing from 87 ± 36 pores/mm(2) at 0% strain to 342 ± 71 at 10% strain; two of the three cell lines showed no additional increase in pore density beyond 10% strain. Transcellular "I-pores" and paracellular "B-pores" both increased with strain (p < 0.038), however B-pores represented the majority (76%) of pores. Pore diameter, in contrast, appeared unaffected by strain (p = 0.25), having a mean diameter of 0.40 μm for I-pores (N = 79 pores) and 0.67 μm for B-pores (N = 350 pores). Pore formation appears to be a mechanosensitive process that is triggered by biomechanical strain, suggesting that SC cells have the ability to modulate local pore density and filtration characteristics of the inner wall endothelium based on local biomechanical cues. The molecular mechanisms of pore formation and how they become altered in glaucoma may be studied in vitro using stretched SC cells.


Associations Between Behavioral Effects of Bisphenol A and DNA Methylation in Zebrafish Embryos.
Endocrine-disrupting contaminants have been associated with aberrant changes in epigenetic pathways in animals. In this study, zebrafish embryos were exposed bisphenol A (BPA) to search for associations between behavior and epigenetic mechanisms in fish. For concentration-dependent responses, embryos were exposed to a range of BPA concentrations (0.1 nM to 30 μM). Embryos were analyzed for locomotor activity at 3-, 4-, and 5-days post fertilization (dpf) in response to changing light conditions. Based on concentration-dependent effects on behavior and gene expression, 10 μM BPA [from 24 to 96 hours post fertilization (hpf)] was used for a whole-genome bisulfite sequencing (WGBS) study searching for genome-wide impacts on DNA methylation. Over the examined concentration ranges, hyperactivity was demonstrated for exposures to 0.001 μM BPA in comparison to embryos exposed to lower or higher BPA concentrations. Transcriptional analysis showed significant effects at >0.01 μM BPA for two genes related to DNA methylation (dnmt1, cbs). BPA exposure did not significantly affect global DNA methylation, but 20,474 differentially methylated (DM) sites in 4,873 genes were identified by WGBS analysis. Most DM sites were identified within gene bodies. The genes with the most DM sites were all protocadherin 2 gamma subfamily genes, related to axon targeting, synaptic development and neuronal survival. KEGG pathways most significantly affected by BPA exposure were phosphatidylinositol signaling system, followed by VEGF and MAPK signaling pathways. This study shows that BPA can affect zebrafish embryo swimming activity at very low concentrations as well as affecting numerous methylated sites in genes which are overrepresented in functionally relevant metabolic pathways. In conclusion, altered methylation patterns of genes associated with nervous system development might lead to abnormal swimming activity.


Influence of airway wall stiffness and parenchymal tethering on the dynamics of bronchoconstriction.
Understanding how tissue remodeling affects airway responsiveness is of key importance, but experimental data bearing on this issue remain scant. We used lung explants to investigate the effects of enzymatic digestion on the rate and magnitude of airway narrowing induced by acetylcholine. To link the observed changes in narrowing dynamics to the degree of alteration in tissue mechanics, we compared our experimental results with predictions made by a computational model of a dynamically contracting elastic airway embedded in elastic parenchyma. We found that treatment of explanted airways with two different proteases (elastase and collagenase) resulted in differential effects on the dynamics of airway narrowing following application of ACh. Histological corroboration of these different effects is manifest in different patterns of elimination of collagen and elastin from within the airway wall and the surrounding parenchyma. Simulations with a computational model of a dynamically contracting airway embedded in elastic parenchyma suggest that elastase exerts its functional effects predominately through a reduction in parenchymal tethering, while the effects of collagenase are more related to a reduction in airway wall stiffness. We conclude that airway and parenchymal remodeling as a result of protease activity can have varied effects on the loads opposing ASM shortening, with corresponding consequences for airway responsiveness.


Patterns of Neuropsychological Profile and Cortical Thinning in Parkinson's Disease with Punding.
Punding, one of dopamine replacement treatment related complications, refers to aimless and stereotyped behaviors. To identify possible neural correlates of punding behavior in patients with Parkinson's disease (PD), we investigated the patterns of cognitive profiles and cortical thinning. Of the 186 subjects with PD screened during the study period, we prospectively enrolled 10 PD patients with punding and 43 without punding on the basis of a structured interview. We performed comprehensive neuropsychological tests and voxel-based and regions-of-interest (ROIs)-based cortical thickness analysis between PD patients with and without punding. The prevalence of punding in patients with PD was 5.4%. Punding behaviors were closely related to previous occupations or hobbies and showed a temporal relationship to changes of levodopa-equivalent dose (LED). Significant predisposing factors were a long duration of PD and intake of medications of PD, high total daily LED, dyskinesia, and impulse control disorder. Punding severity was correlated with LED (p = 0.029). The neurocognitive assessment revealed that PD patients with punding showed more severe cognitive deficits in the color Stroop task than did those without punding (p = 0.022). Voxel-based analysis showed that PD-punders had significant cortical thinning in the dorsolateral prefrontal area relative to controls. Additionally, ROI-based analysis revealed that cortical thinning in PD-punders relative to PD-nonpunders was localized in the prefrontal cortices, extending into orbitofrontal area. We demonstrated that PD patients with punding performed poorly on cognitive tasks in frontal executive functions and showed severe cortical thinning in the dorsolateral prefrontal and orbitofrontal areas. These findings suggest that prefrontal modulation may be an essential component in the development of punding behavior in patients with PD.


NOD/SCID IL2Rγ-null mouse xenograft model of human p53-mutated chronic lymphocytic leukemia and ATM-mutated mantle cell lymphoma using permanent cell lines.
Xenograft models represent a promising tool to study the pathogenesis of hematological malignancies. To establish a reliable and appropriate in vivo model of aggressive human B-cell leukemia and lymphoma we xenotransplanted four p53-mutated cell lines and one ATM-mutated cell line into immunodeficient NOD/SCID IL2Rγ-null mice. The cell lines MEC-1, SU-DHL-4, JEKO-1, REC-1, and GRANTA-519 were transplanted intraperitoneally or subcutaneously and the engraftment was investigated using immunohistochemistry and flow cytometry. We found significant differences in engraftment efficiency. MEC-1, JEKO-1 and GRANTA-519 cell lines engrafted most efficiently, while SU-DHL-4 cells did not engraft at all. MEC-1 and GRANTA-519 massively infiltrated organs and the whole intraperitoneal cavity showing very aggressive growth. In addition, GRANTA-519 cells massively migrated to the bone marrow regardless of the transplantation route. The MEC-1 and GRANTA-519 cells can be especially recommended for in vivo study of p53-mutated chronic lymphocytic leukemia and ATM-mutated mantle cell lymphoma, respectively.


Large-scale immuno-magnetic cell sorting of T cells based on a self-designed high-throughput system for potential clinical application.
In this contribution, we designed four types of immuno-magnetic nanoparticles for separation of different T cells (CD3+, CD4+, CD8+ and CD14+ T cells), and we established a new large-scale immuno-magnetic cell sorting procedure to achieve an enrichment of particular T cells using our designed auto-IMACS device. This device could achieve recyclable large-scale cell sorting, for which the throughput of the system reached ∼4000 mL and the maximum cell capacity was 4 × 1010. The collected cells were analyzed by flow cytometry and visual cytology data, and the effective selection rates of CD3+, CD4+, CD8+ and CD14+ T cells were 79.3%, 74.1%, 57.1% and 67.9%, respectively. The sorted CD8+ T cells still retained good cytotoxic activity against specific cells. In addition, the sorted T cells can also be further incubated in vitro and proliferated, and even could be infused back into patients for immunotherapy in the near future.


Carrier dynamics and spin-valley-layer effects in bilayer transition metal dichalcogenides.
Transition metal dichalcogenides are an interesting class of low dimensional materials in mono- and few-layer form with diverse applications in valleytronic, optoelectronic and quantum devices. Therefore, the general nature of the band-edges and the interplay with valley dynamics is important from a fundamental and technological standpoint. Bilayers introduce interlayer coupling effects which can have a significant impact on the valley polarization. The combined effect of spin-orbit and interlayer coupling can strongly modify the band structure, phonon interactions and overall carrier dynamics in the material. Here we use first-principles calculations of electron-electron and electron-phonon interactions to investigate bilayer MoS2 and WSe2 in both the AA' and AB stacking configurations. We find that in addition to spin-orbit coupling, interlayer interactions present in the two configurations significantly alter the near-band-edge dynamics. Scattering lifetimes and dynamic behavior are highly material-dependent, despite the similarities and typical trends in TMDCs. Additionally, we capture significant differences in dynamics for the AA' and AB stacking configurations, with lifetime values differing by up to an order of magnitude between them for MoS2. Further, we evaluate the valley polarization times and find that maximum lifetimes at room temperature are of the scale of 1 picosecond for WSe2 in the AB orientation. These results present a pathway to understanding complex heterostructure configurations and 'magic angle' physics in TMDCs.


Fever as the presenting complaint of travellers returning from the tropics.
We investigated prospectively the cause of fever in patients requiring hospitalization after returning from the tropics. All consecutive admissions (n = 195) with oral temperature > 37.0 degrees C at the time of admission were enrolled. Final diagnosis as recorded on the discharge summary by the attending physician and results of any relevant laboratory or radiological investigations were recorded on standard proforma. Malaria accounted for 42% of admissions; two patients had returned to Britain more than 6 months before presentation. The second largest group was assumed to have a non-specific viral infection (25%). Cosmopolitan infections (urinary tract infection, community-acquired pneumonia, streptococcal sore throat, etc.) accounted for 9%. Coincidental infections (schistosomiasis, filariasis, intestinal helminths) were found in 16%. Serology was positive for HIV infection in 3%. The most useful investigation was a malaria film, which was positive in 45% of cases in which it was performed. The combination of thrombocytopaenia (platelet count < 100 x 10(9)) and hyperbilirubinaemia (bilirubin > 18 IU/ml) were useful predictive markers of malaria: all 23 patients with both abnormalities had positive malaria films. Malaria must be excluded in any febrile patient returning from the tropics. In the absence of a positive malaria film, the combination of a low platelet count and raised bilirubin may suggest the need for an empirical course of therapy.


What is the meaning of a 'genomic result' in the context of pregnancy?
Prenatal genetic testing and analysis in the past was usually only offered when a particular fetal phenotype was noted or suspected, meaning that filtering and interpretation of genetic variants identified could be anchored in attempts to explain an existing health concern. Advanced genomic testing is now increasingly used in "low-risk" pregnancies, producing information on genotype adrift of the phenotypic data that is necessary to give it meaning, thus increasing the difficulty in predicting whether and how particular genetic variants might affect future development and health. A challenge to healthcare scientists, clinicians, and parents therefore is deciding what qualities prenatal genotypic variation should have in order to be constructed as a 'result.' At the same time, such tests are often re requested in order to make binary decisions about whether to continue a pregnancy or not. As a range of professional organizations develop guidelines on the use of advanced genomic testing during pregnancy, we highlight the particular difficulties of discovering ambiguous findings such as variants with uncertain clinical significance, susceptibility loci for neurodevelopmental problems and susceptibility to adult-onset diseases. We aim to foster international discussions about how decisions around disclosure are made and how uncertainty is communicated.


Controlling the spread of carbapenemase-producing Gram-negatives: therapeutic approach and infection control.
Although the rapid spread of carbapenemase-producing Gram-negatives (CPGNs) is providing the scientific community with a great deal of information about the molecular epidemiology of these enzymes and their genetic background, data on how to treat multidrug-resistant or extended drug-resistant carbapenemase-producing Enterobacteriaceae and how to contain their spread are still surprisingly limited, in spite of the rapidly increasing prevalence of these organisms and of their isolation from patients suffering from life-threatening infections. Limited clinical experience and several in vitro synergy studies seem to support the view that antibiotic combinations should be preferred to monotherapies. But, in light of the data available to date, it is currently impossible to quantify the real advantage of drug combinations in the treatment of these infections. Comprehensive clinical studies of the main therapeutic options, broken down by pathogen, enzyme and clinical syndrome, are definitely lacking and, as carbapenemases keep spreading, are urgently needed. This spread is unveiling the substantial unpreparedness of European public health structures to face this worrisome emergency, although experiences from different countries-chiefly Greece and Israel-have shown that CPGN transmission and cross-infection can cause a substantial threat to the healthcare system. This unpreparedness also affects the treatment of individual patients and infection control policies, with dramatic scarcities of both therapeutic options and infection control measures. Although correct implementation of such measures is presumably cumbersome and expensive, the huge clinical and public health problems related to CPGN transmission, alongside the current scarcity of therapeutic options, seem to fully justify this choice.


Long-term disability associated with war-related experience among Vietnam veterans: retrospective cohort study.
Recent combat operations have involved large numbers of personnel. Long-term health effects of military deployment remain largely unknown. To examine patterns and trends in long-term disability among combat veterans and to relate disability to aspects of wartime experience. A total of 60,228 Australian military personnel deployed between 1962 and 1975 during the Vietnam War, and 82,877 military personnel who were not deployed overseas. Accepted physician-assessed disability claims were evaluated over follow-up periods up to 50 years after deployment, and compared with age-matched controls. Multivariable analysis was used to examine differences by service branch, rank, age, and deployment duration. The steepest rise in disability incidence was observed among Vietnam veterans starting in the 1990s, around 20-30 years after deployment for most veterans. After 1994, when Statements of Principles were introduced to guide evaluation of disability claims, the hazard ratio for disability incidence was 1.53 (95% confidence interval, 1.32-1.77) compared with the prior period. By January 2011, after an average follow-up of 42.5 years, 69.7% (95% confidence interval, 69.4%-70.1%) of veterans had at least 1 war-related disability. Many veterans had multiple disabilities, with leading causes being eye and ear disorders (48.0%), mental health conditions (47.9%), and musculoskeletal disorders (18.4%). For specific categories of disability, relative risks for accepted claims among veterans compared with controls were highest for mental health disorders, at 22.9 (21.9-24.0) and lowest for injuries, at 1.5 (1.4-1.6) with a relative risk for any disability of 3.7 (3.7-3.8). Veterans with service of >1 year were 2.5 (2.2-2.7) times more likely to have a mental health disability than those who served <100 days, and 2.3 (2.1-2.5) times more likely to have other disabilities. Long-term effects of deployment into military conflicts are substantial, and likelihood of war-related disability is associated with service history. If similar patterns follow from more recent conflicts, significant additional resources will be needed to prevent and treat long-term health conditions among veterans.


Phenomenology of prolonged febrile seizures: results of the FEBSTAT study.
Febrile status epilepticus (FSE) has been associated with hippocampal injury and subsequent mesial temporal sclerosis and temporal lobe epilepsy. However, little is known about the semiology of FSE. A prospective, multicenter study of the consequences of FSE included children, aged 1 month through 5 years, presenting with a febrile seizure lasting 30 minutes or more. Procedures included neurologic history and examination and an MRI and EEG within 72 hours. All information related to seizure semiology was reviewed by three epileptologists blinded to MRI and EEG results and to subsequent outcome. Inter-rater reliability was assessed by the kappa statistic. Among 119 children, the median age was 1.3 years, the mean peak temperature was 103.2 degrees F, and seizures lasted a median of 68.0 minutes. Seizure duration followed a Weibull distribution with a shape parameter of 1.68. Seizures were continuous in 52% and behaviorally intermittent (without recovery in between) in 48%; most were partial (67%) and almost all (99%) were convulsive. In one third of cases, FSE was unrecognized in the emergency department. Of the 119 children, 86% had normal development, 24% had prior febrile seizures, and family history of febrile seizures in a first-degree relative was present in 25%. Febrile status epilepticus is usually focal and often not well recognized. It occurs in very young children and is usually the first febrile seizure. Seizures are typically very prolonged and the distribution of seizure durations suggests that the longer a seizure continues, the less likely it is to spontaneously stop.


Elucidating the kinetic mechanism of DNA polymerization catalyzed by Sulfolobus solfataricus P2 DNA polymerase B1.
Transient-state kinetic techniques were used to resolve the kinetic mechanism of DNA polymerization catalyzed by an exonuclease-deficient mutant of Sulfolobus solfataricus P2 DNA polymerase B1 (PolB1 exo-). Here, we report the kinetic parameters of several elementary steps for the forward polymerization reaction. PolB1 exo- binds tightly to DNA (K(d)(DNA) = 1.8 nM) and a correct incoming nucleotide (apparent K(d)(dTTP) = 11 microM). Moreover, several lines of kinetic evidence suggested that correct nucleotide incorporation catalyzed by PolB1 exo- was limited by a protein conformational change which precedes the chemistry step. The utilization of an "induced fit" mechanism by PolB1 exo- was supported by the following: a small, alpha-thio elemental effect of 1.5, varying DNA dissociation rates for the binary complex (0.043 s(-1)) as well as ternary complexes before (0.18 s(-1)) and after (0.0071 s(-1)) a conformational change, a greater amplitude for the pulse-chase than the pulse-quench reaction, and an activation energy barrier of 38 kcal/mol which is greater than the predicted values of phosphodiester bond formation both in solution and within a polymerase active site. Lastly, PolB1 exo- exhibited a low processivity value of 15, thereby suggesting a protein cofactor confers this replicative DNA polymerase with higher processivity in vivo.


Oxysterol Sensing through the Receptor GPR183 Promotes the Lymphoid-Tissue-Inducing Function of Innate Lymphoid Cells and Colonic Inflammation.
Group 3 innate lymphoid cells (ILC3s) sense environmental signals and are critical for tissue integrity in the intestine. Yet, which signals are sensed and what receptors control ILC3 function remain poorly understood. Here, we show that ILC3s with a lymphoid-tissue-inducer (LTi) phenotype expressed G-protein-coupled receptor 183 (GPR183) and migrated to its oxysterol ligand 7α,25-hydroxycholesterol (7α,25-OHC). In mice lacking Gpr183 or 7α,25-OHC, ILC3s failed to localize to cryptopatches (CPs) and isolated lymphoid follicles (ILFs). Gpr183 deficiency in ILC3s caused a defect in CP and ILF formation in the colon, but not in the small intestine. Localized oxysterol production by fibroblastic stromal cells provided an essential signal for colonic lymphoid tissue development, and inflammation-induced increased oxysterol production caused colitis through GPR183-mediated cell recruitment. Our findings show that GPR183 promotes lymphoid organ development and indicate that oxysterol-GPR183-dependent positioning within tissues controls ILC3 activity and intestinal homeostasis.


Numerical simulation of millisecond laser-induced damage in silicon-based positive-intrinsic-negative photodiode.
An axisymmetric mathematical model was established for millisecond-pulsed Nd:YAG laser heating of silicon-based positive-intrinsic-negative photodiode. The transient temperature fields were obtained by using the finite element method. The temperature dependences of the material parameters and the absorption coefficient were taken into account in the calculation. The results indicate that the optical absorption coefficient and the thermal conductivity are the two key factors for the temperature evolution. The diffusion of boron in the liquid phase and the introduction of deep-level defects in the depletion region of the photodiode were the two reasons for the millisecond laser-induced electrical degradation of the photodiode. The morphological damage threshold and electrical degradation threshold of the photodiode were obtained numerically. Meanwhile, the influence of the antireflection coating, the doping concentration, and the junction depth were also considered. The results show that the morphological damage threshold decreases with adding an antireflection coating, the increase of the doping concentration, and junction depth. The electrical degradation threshold increases only with the junction depth.


Alternative approaches for efficient inhibition of hepatitis C virus RNA replication by small interfering RNAs.
Persistent infection with hepatitis C virus (HCV) is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. It has recently been shown that HCV RNA replication is susceptible to small interfering RNAs (siRNAs), but the antiviral activity of siRNAs depends very much on their complementarity to the target sequence. Thus, the high degree of sequence diversity between different HCV genotypes and the rapid evolution of new quasispecies is a major problem in the development of siRNA-based gene therapies. For this study, we developed two alternative strategies to overcome these obstacles. In one approach, we used endoribonuclease-prepared siRNAs (esiRNAs) to simultaneously target multiple sites of the viral genome. We show that esiRNAs directed against various regions of the HCV coding sequence as well as the 5' nontranslated region (5' NTR) efficiently block the replication of subgenomic and genomic HCV replicons. In an alternative approach, we generated pseudotyped retroviruses encoding short hairpin RNAs (shRNAs). A total of 12 shRNAs, most of them targeting highly conserved sequence motifs within the 5' NTR or the early core coding region, were analyzed for their antiviral activities. After the transduction of Huh-7 cells containing a subgenomic HCV replicon, we found that all shRNAs targeting sequences in domain IV or nearby coding sequences blocked viral replication. In contrast, only one of seven shRNAs targeting sequences in domain II or III had a similar degree of antiviral activity, indicating that large sections of the NTRs are resistant to RNA interference. Moreover, we show that naive Huh-7 cells that stably expressed certain 5' NTR-specific shRNAs were largely resistant to a challenge with HCV replicons. These results demonstrate that the retroviral transduction of HCV-specific shRNAs provides a new possibility for antiviral intervention.


Abscess induction in beige (bg/bg) mutant mice.
Humans with Chediak-Higashi Syndrome (CHS) and several animal species with similar defects have been reported to have increased susceptibility to infection, including abscess formation, associated with granulocyte abnormalities. These defects were investigated by comparing mice of the SB/Le strain, homozygous for the beige mutation, with their heterozygous littermates and with normal BALB/c mice with respect to the ability to form intraabdominal abscesses. Mice were autopsied 7 days after the intraperitoneal inoculation of a mixture of Bacteroides fragilis, Escherichia coli and an abscess-potentiating agent, bran. Although homozygous beige mice formed more numerous abscesses than controls, the total abscess sizes and bacterial contents were not significantly different. Histopathologically, the abscesses resembled those in normal mice. Phagocytic killing assays using granulocytes from beige mice of the SB/Le and C57BL/6J strains showed no significant differences between homozygous beige mice and controls. Within the context of these experiments in which neutrophil function in vivo and in vitro was examined with respect to anaerobic and facultative Gram-negative bacteria, it is concluded that beige mice can respond adequately to an infectious bacterial challenge. The increased susceptibility to an infection seen in various species with CHS-like disease may relate to virulence factors of the infecting organisms and defects in host defences documented by others but not manifest in these experiments.


Determination of mineral, trace element, and pesticide levels in honey samples originating from different regions of Malaysia compared to manuka honey.
The present study was undertaken to determine the content of six minerals, five trace elements, and ten pesticide residues in honeys originating from different regions of Malaysia. Calcium (Ca), magnesium (Mg), iron (Fe), and zinc (Zn) were analyzed by flame atomic absorption spectrometry (FAAS), while sodium (Na) and potassium (K) were analyzed by flame emission spectrometry (FAES). Trace elements such as arsenic (As), lead (Pb), cadmium (Cd), copper (Cu), and cobalt (Co) were analyzed by graphite furnace atomic absorption spectrometry (GFAAS) following the microwave digestion of honey. High mineral contents were observed in the investigated honeys with K, Na, Ca, and Fe being the most abundant elements (mean concentrations of 1349.34, 236.80, 183.67, and 162.31 mg/kg, resp.). The concentrations of the trace elements were within the recommended limits, indicating that the honeys were of good quality. Principal component analysis reveals good discrimination between the different honey samples. The pesticide analysis for the presence of organophosphorus and carbamates was performed by high performance liquid chromatography (HPLC). No pesticide residues were detected in any of the investigated honey samples, indicating that the honeys were pure. Our study reveals that Malaysian honeys are rich sources of minerals with trace elements present within permissible limits and that they are free from pesticide contamination.


The influence of HIV status on prenatal genetic diagnosis choices.
At-risk women of advanced maternal age (AMA) can choose to have second-trimester invasive testing for a prenatal genetic diagnosis on the fetus. Being HIV-positive can complicate the decision-making process. To document HIV status and prenatal genetic diagnosis choices in women of AMA attending genetic clinics in Johannesburg, South Africa, for counselling on the risks of abnormalities in their fetuses. Data on the characteristics of the sample, HIV status and prenatal diagnosis decisions were collected retrospectively from the files of 350 women (>34 years) counselled for AMA in genetic clinics in Johannesburg and Pretoria. The time period was 6 months in 2003 and 6 months in 2004. A sample of the women (n=15) who were HIV-positive were interviewed and completed questionnaires on their understanding of their situation. The data were analysed and submitted to statistical testing. Of the 350 women, 183 (52.3%) were HIV-negative, 44 (12.6%) HIV-positive, and 123 (35.1%) of unknown status. Significantly more HIV-negative patients (79/183, 43.2%) than those who were HIV-positive (6/44, 13.6%) had amniocentesis performed for fetal diagnosis. Most of the interviewed women (12/15, 80.0%) understood the severity of HIV infection, 5 (33.3%) considered termination of pregnancy based on the transmission risk, and 4 (26.7%) would have requested amniocentesis and prenatal diagnosis if they had been HIV-negative. CONCLUSION. Decision-making regarding prenatal genetic diagnosis is influenced by HIV status among older women. Effective access to highly active antiretroviral therapy throughout pregnancy would make decision-making easier for these women.


Relation between clinical efficacy and pharmacokinetic parameters after sublingual apomorphine in Parkinson's disease.
Apomorphine was administered sublingually in two single doses (0.3 and 0.6 mg/kg) to seven patients with idiopathic Parkinson's disease (PD) to assess the relation between clinical efficacy, dosage, and pharmacokinetic parameters of apomorphine. On day 1 and day 3, patients were given 0.3 mg/kg and 0.6 mg/kg of apomorphine, respectively (3 mg tablets). Before apomorphine administration and during the following 4 h, motor score was assessed by measuring tremor, akinesia scores, rising from a chair, and walking speed. The delay to turn on was not different between the two doses but after the 0.3 mg/kg dose, only three patients turned on, whereas all the patients treated with 0.6 mg/kg turned on. Apomorphine (0.3 mg/kg) induced a shorter duration of the "on" period than 0.6 mg/kg (0.3 mg/kg: 24.2 +/- 14.6 min; 0.6 mg/kg: 86.7 +/- 14.9 min). The time to obtain the peak plasma concentration (tmax) obtained with the two doses were not different (0.3 mg/kg: 31.5 +/- 3.4 min; 0.6 mg/kg: 38.3 +/- 2.8 min). Peak plasma concentrations (Cmax) and areas under the curve (AUC) were significantly higher after 0.6 mg/kg than 0.3 mg/kg (Cmax: 0.3 mg/kg: 7.5 +/- 3.2 ng/ml; 0.6 mg/kg: 22.7 +/- 3.6 ng/ml; p < 0.01; AUC: 0.3 mg/kg: 929 +/- 109 ng/ml/min; 0.6 mg/kg; 2,277 +/- 209 ng/ml/min; p < 0.01). There was a significant linear correlation between the duration of therapeutic effect, AUC, and Cmax (r = 0.86, p < 0.01 for AUC; r = 0.63, p < 0.05 for Cmax). These results show that sublingual apomorphine could be of interest in the treatment of "off" phases in parkinsonian patients with motor fluctuations.


Prevalence of cocaine use in Brazil: data from the II Brazilian national alcohol and drugs survey (BNADS).
To determine the current prevalence rates of cocaine use and dependence in a representative sample of the Brazilian population, and to investigate possible associations with sociodemographic factors. The Second Brazilian Alcohol and Drugs Survey (II BNADS) interviewed 4607 individuals aged 14 years and older from the Brazilian household population including an over sample of 1157 adolescents (14 to 18 years old).The survey gathered information on alcohol, tobacco and illegal substances use as well as on risk factors for abuse and dependence. The Severity of Dependence Scale was used to evaluate cocaine dependence rates among users. Last year consumption of crack-cocaine was 2.2% in the overall population excluding the elderly group.Lifetime and last year prevalence rate of snorted cocaine was 3.9% and 1.7%, respectively. Smoked cocaine use in Brazil was estimated in 1.5% for lifetime and 0.8% for last year use. Cocaine addiction was identified in 41.4%between users in the prior year. The prevalence rates of snorted and smoked cocaine in Brazil suggests that the country is amongst the nations with greatest annual consumption rates becoming one of the biggest consumer markets of cocaine worldwide. Prevention and treatment policies should take this into consideration and strengthen the focus on cocaine use in the country.


Characterisation of phenolics, betanins and antioxidant activities in seeds of three Chenopodium quinoa Willd. genotypes.
Quinoa (Chenopodium quinoa Willd.) is known for its exceptional nutritional value and potential health benefits. The present study identified the composition of different forms of extractable phenolics and betacyanins of quinoa cultivars in white, red and black, and how they contribute to antioxidant activities. Results showed that at least 23 phenolic compounds were found in either free or conjugated forms (liberated by alkaline and/or acid hydrolysis); the majority of which were phenolic acids, mainly vanillic acid, ferulic acid and their derivatives as well as main flavonoids quercetin, kaempferol and their glycosides. Betacyanins, mainly betanin and isobetanin, were confirmed for the first time to be the pigments of the red and black quinoa seeds, instead of anthocyanins. Darker quinoa seeds had higher phenolic concentration and antioxidant activity. Findings of these phenolics, along with betacyanins in this study add new knowledge to the functional components of quinoa seeds of different cultivar background.


Hg-responsive proteins identified in wheat seedlings using iTRAQ analysis and the role of ABA in Hg stress.
Wheat seedlings exposed to 100 μM HgCl2 for 3 days exhibited high-level mercury (Hg) accumulation, which led to inhibited growth, increased lipid peroxidation, and disrupted cellular ultrastructures. And root growth and ultrastructural changes of wheat seedlings were inhibited more severely than those of leaves. To identify the wheat protein response to Hg stress, the iTRAQ method was used to determine the proteome profiles of the roots and leaves of wheat seedlings exposed to high-Hg conditions. 249 proteins were identified with significantly altered abundance. 117 were found in roots and 132 in leaves. These proteins were classified into signal transduction, stress defense, carbohydrate metabolism, protein metabolism, energy production, and transport functional groups. The majority of proteins identified in Hg-stressed roots and leaves displayed differently altered abundance, revealing organ-specific differences in adaption to Hg stress. Pathway Studio software was used to identify the Hg-responsive protein interaction network that included 49 putative key proteins, and they were potentially regulated by abscisic acid (ABA). Exogenous ABA application conferred protection against Hg stress and increased activities of peroxidase enzyme, suggesting that it may be an important factor in the Hg signaling pathway. These findings can provide useful insights into the molecular mechanisms of Hg responses in higher plants.


Pediatric desmoid tumor: retrospective analysis of 63 cases.
This study was conducted to evaluate clinical prognostic factors predictive of the probability of recurrence of desmoid tumor (DT). Sixty-three patients with histologically confirmed diagnosis of DT were retrospectively studied. Median age at diagnosis was 13 years. Patient distribution by site was as follows: 61% extremities, 18% head and neck, 13% trunk (including 5% intraabdominal), and 8% multicentric lesions. All patients had partial or complete resections; 20 patients also received radiotherapy and/or chemotherapy. At a median follow-up time of 6 years since first treatment, the overall actuarial probability of having one or more recurrences was 75%. Age, sex, site, size, or number of previous recurrences had no significant impact on the likelihood of recurrence. The only factor associated with an increased proportion of recurrence-free patients was a negative margin of resection (70% v 15% with positive margins; P = .006). Of the four patients with more than 3 years follow-up since chemotherapy, two recurred, and of the 11 patients with the same follow-up after radiotherapy, four recurred, including two of five patients who received a dose of 50 Gy or more. No deaths directly related to tumor invasion were observed. A surgical approach aiming at clear margins is presently the best treatment option. When this cannot be accomplished without severe disfigurement or function impairment, partial resection is an acceptable alternative, but one associated with a high risk of regrowth. Whether adjuvant strategies should be used in this situation remains to be addressed.


Bioluminescent response of individual dinoflagellate cells to hydrodynamic stress measured with millisecond resolution in a microfluidic device.
Dinoflagellate bioluminescence serves as a model system for examining mechanosensing by suspended motile unicellular organisms. The response latency, i.e. the delay time between the mechanical stimulus and luminescent response, provides information about the mechanotransduction and signaling process, and must be accurately known for dinoflagellate bioluminescence to be used as a flow visualization tool. This study used a novel microfluidic device to measure the response latency of a large number of individual dinoflagellates with a resolution of a few milliseconds. Suspended cells of several dinoflagellate species approximately 35 microm in diameter were directed through a 200 microm deep channel to a barrier with a 15 microm clearance impassable to the cells. Bioluminescence was stimulated when cells encountered the barrier and experienced an abrupt increase in hydrodynamic drag, and was imaged using high numerical aperture optics and a high-speed low-light video system. The average response latency for Lingulodinium polyedrum strain HJ was 15 ms (N>300 cells) at the three highest flow rates tested, with a minimum latency of 12 ms. Cells produced multiple flashes with an interval as short as 5 ms between individual flashes, suggesting that repeat stimulation involved a subset of the entire intracellular signaling pathway. The mean response latency for the dinoflagellates Pyrodinium bahamense, Alexandrium monilatum and older and newer isolates of L. polyedrum ranged from 15 to 22 ms, similar to the latencies previously determined for larger dinoflagellates with different morphologies, possibly reflecting optimization of dinoflagellate bioluminescence as a rapid anti-predation behavior.


Temporal efficacy of allopurinol during the induction of pancreatitis in the ex vivo perfused canine pancreas.
Oxygen-derived free radicals play an important role in the pathogenesis of experimental acute pancreatitis in the isolated perfused canine pancreas. We have previously found that pretreatment with allopurinol inhibits xanthine oxidase--apparently the primary source of free radical generation in this model--and prevents the initial development of pancreatitis. In these experiments, we evaluated whether allopurinol administered after the onset of pancreatitis would arrest the progression of the disease process. Edema formation, weight gain, and the release of amylase activity into the perfusate in the ex vivo perfused canine pancreas model were monitored during a 4-hour perfusion period. There were six experimental groups: Group I (control) received no treatment, group II (allopurinol alone) received only allopurinol (100 mg) at the start of perfusion, and groups III through VI were each given an infusion of 0.3 ml of oleic acid (FFA) over a 1-hour period to initiate acute pancreatitis. Group III (FFA alone) received no other treatment. In group IV (pretreatment with allopurinol), group V (concurrent treatment with allopurinol), and group VI (posttreatment with allopurinol), allopurinol (100 mg) was administered 1 hour before, concurrent with, or at the end of the FFA infusion, respectively. Pretreatment with allopurinol prevented edema formation, markedly attenuated weight gain, and the release of amylase caused by the FFA infusion. Administration of allopurinol concurrent with the FFA infusion provided only partial protection, whereas posttreatment with allopurinol failed to arrest the progression of the injury process. Therefore, the use of allopurinol to inhibit oxygen-derived free radical production from xanthine oxidase prevented the development of acute pancreatitis in this model; however, treatment with allopurinol after initiation of the disease process failed to arrest the progression of acute pancreatitis.


Measuring drug absorption improves interpretation of behavioral responses in a larval zebrafish locomotor assay for predicting seizure liability.
Unanticipated effects on the central nervous system are a concern during new drug development. A larval zebrafish locomotor assay can reveal seizure liability of experimental molecules before testing in mammals. Relative absorption of compounds by larvae is lacking in prior reports of such assays; having those data may be valuable for interpreting seizure liability assay performance. Twenty-eight reference drugs were tested at multiple dose levels in fish water and analyzed by a blinded investigator. Responses of larval zebrafish were quantified during a 30min dosing period. Predictive metrics were calculated by comparing fish activity to mammalian seizure liability for each drug. Drug level analysis was performed to calculate concentrations in dose solutions and larvae. Fifteen drug candidates with neuronal targets, some having preclinical convulsion findings in mammals, were tested similarly. The assay has good predictive value of established mammalian responses for reference drugs. Analysis of drug absorption by larval fish revealed a positive correlation between hyperactive behavior and pro-convulsive drug absorption. False negative results were associated with significantly lower compound absorption compared to true negative, or true positive results. The predictive value for preclinical toxicology findings was inferior to that suggested by reference drugs. Disproportionately low exposures in larvae giving false negative results demonstrate that drug exposure analysis can help interpret results. Due to the rigorous testing commonly performed in preclinical toxicology, predicting convulsions in those studies may be more difficult than predicting effects from marketed drugs.


Electrophilic natural products and their biological targets.
The study of biologically active natural products has resulted in seminal contributions to our understanding of living systems. In the case of electrophilic natural products, the covalent nature of their interaction has largely facilitated the identification of their biological binding partners. In this review, we provide a comprehensive compilation of electrophilic natural products from all major chemical classes together with their biological targets. Covering Michael acceptor systems, ring-strained compounds and other electrophiles, such as esters or carbamates, we highlight representative and instructive examples for over 20 electrophilic moieties. The fruitful cooperation of natural product chemistry, medicinal chemistry and chemical biology has produced a collection of well-studied examples for how electrophilic natural products exert their biological functions that range from antibiotic to antitumor effects. Special emphasis is put on the elucidation of their respective biological targets via activity-based protein profiling, which together with the recent advancements in mass spectrometry has been crucial to the success of the field. The wealth of naturally occurring electrophilic moieties and their chemical complexity enables binding of a large variety of biological targets, such as enzymes of all classes, nonenzymatic proteins, DNA and other cellular compounds. With approximately 30,000 genes in the human genome but only 266 confirmed protein drug targets, the study of biologically active, electrophilic natural products has the potential to provide insights into fundamental biological processes and to greatly aid the discovery of new drug targets.


The epidemiology of 5-methoxy- N, N-dimethyltryptamine (5-MeO-DMT) use: Benefits, consequences, patterns of use, subjective effects, and reasons for consumption.
5-Methoxy- N,N-dimethyltryptamine (5-MeO-DMT) is a psychoactive compound found in several plants and in high concentrations in Bufo alvarius toad venom. Synthetic, toad, and plant-sourced 5-MeO-DMT are used for spiritual and recreational purposes and may have psychotherapeutic effects. However, the use of 5-MeO-DMT is not well understood. Therefore, we examined patterns of use, motivations for consumption, subjective effects, and potential benefits and consequences associated with 5-MeO-DMT use. Using internet-based advertisements, 515 respondents ( Mage=35.4. SD=11.7; male=79%; White/Caucasian=86%; United States resident=42%) completed a web-based survey. Most respondents consumed 5-MeO-DMT infrequently (<once/year), for spiritual exploration, and had used less than four times in their lifetime. The majority (average of 90%) reported moderate-to-strong mystical-type experiences ( Mintensity=3.64, SD=1.11; range 0-5; e.g., ineffability, timelessness, awe/amazement, experience of pure being/awareness), and relatively fewer (average of 37%) experienced very slight challenging experiences ( Mintensity=0.95, SD=0.91; range 0-5; e.g., anxiousness, fear). Less than half (39%) reported repeated consumption during the same session, and very few reported drug craving/desire (8%), or legal (1%), medical (1%), or psychiatric (1%) problems related to use. Furthermore, of those who reported being diagnosed with psychiatric disorders, the majority reported improvements in symptoms following 5-MeO-DMT use, including improvements related to post-traumatic stress disorder (79%), depression (77%), anxiety (69%), and alcoholism (66%) or drug use disorder (60%). Findings suggest that 5-MeO-DMT is used infrequently, predominantly for spiritual exploration, has low potential for addiction, and might have psychotherapeutic effects. Future research should examine the safety and pharmacokinetics of 5-MeO-DMT administration in humans using rigorous experimental designs.


Secondary Hypertension and Complications: Diagnosis and Role of Imaging.
Hypertension is a common problem; if left untreated, it can result in significant complications, including those involving the cardiovascular system and end organs. Approximately 10% of patients with hypertension are classified as having secondary hypertension, defined as hypertension attributable to a specific and potentially remediable cause. The evaluation for secondary hypertension typically begins with acquiring the patient history and performing a physical examination and screening laboratory tests. Directed imaging may be performed, on the basis of laboratory test results, to assess for potential causes of secondary hypertension. The causes can be broadly classified as endocrine (eg, hyperaldosteronism, pheochromocytoma, hyperparathyroidism) and nonendocrine (eg, aortic coarctation, renal vascular hypertension). In addition, patients with hypertension can develop significant complications that also are diagnosed with imaging, including conditions involving the cardiovascular system (eg, aortic aneurysm, acute aortic syndrome) and central nervous system (eg, stroke, subarachnoid hemorrhage, and posterior reversible encephalopathy syndrome). The imaging workup and imaging appearances of some of the causes of secondary hypertension are reviewed, treatment options are discussed, and the imaging appearances of hypertension-related complications are described. It is important for radiologists to accurately diagnose the secondary causes of hypertension, as many of them are treatable, and treatment may result in improved symptoms or resolution of hypertension. ©RSNA, 2019.


RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome.
RNA-Seq is revolutionizing the way transcript abundances are measured. A key challenge in transcript quantification from RNA-Seq data is the handling of reads that map to multiple genes or isoforms. This issue is particularly important for quantification with de novo transcriptome assemblies in the absence of sequenced genomes, as it is difficult to determine which transcripts are isoforms of the same gene. A second significant issue is the design of RNA-Seq experiments, in terms of the number of reads, read length, and whether reads come from one or both ends of cDNA fragments. We present RSEM, an user-friendly software package for quantifying gene and isoform abundances from single-end or paired-end RNA-Seq data. RSEM outputs abundance estimates, 95% credibility intervals, and visualization files and can also simulate RNA-Seq data. In contrast to other existing tools, the software does not require a reference genome. Thus, in combination with a de novo transcriptome assembler, RSEM enables accurate transcript quantification for species without sequenced genomes. On simulated and real data sets, RSEM has superior or comparable performance to quantification methods that rely on a reference genome. Taking advantage of RSEM's ability to effectively use ambiguously-mapping reads, we show that accurate gene-level abundance estimates are best obtained with large numbers of short single-end reads. On the other hand, estimates of the relative frequencies of isoforms within single genes may be improved through the use of paired-end reads, depending on the number of possible splice forms for each gene. RSEM is an accurate and user-friendly software tool for quantifying transcript abundances from RNA-Seq data. As it does not rely on the existence of a reference genome, it is particularly useful for quantification with de novo transcriptome assemblies. In addition, RSEM has enabled valuable guidance for cost-efficient design of quantification experiments with RNA-Seq, which is currently relatively expensive.


Development of an Intracranial Dural Arteriovenous Fistula after Venous Sinus Stenting for Idiopathic Intracranial Hypertension.
We report a case in which an intracranial dural arteriovenous fistula (DAVF) developed after endovascular treatment of a patient with idiopathic intracranial hypertension with venous sinus stenting (VSS). The pathogenesis may involve hemodynamic alterations secondary to increased poststenting venous sinus pressure, which may cause new arterial ingrowth into the fistulous sinus wall without capillary interposition. Despite administration of dual antiplatelet therapy, there may also be subclinical cortical vein thrombosis that contributed to DAVF formation. In addition to the aforementioned mechanisms, increased inflammation induced by VSS may upregulate vascular endothelial growth factor and platelet-derived growth factor expression and also promote DAVF pathogenesis. Since VSS has been used to obliterate DAVFs, DAVF formation after VSS may seem counterintuitive. Previous stents have generally been closed cell, stainless steel designs used to maximize radial compression of the fistulous sinus wall. In contrast, our patient's stent was an open cell, self-expandable nitinol design (Protégé Everflex). Neurointerventionalists should be aware of this potential, although rare complication of DAVF formation after VSS.


Emo, love and god: making sense of Urban Dictionary, a crowd-sourced online dictionary.
The Internet facilitates large-scale collaborative projects and the emergence of Web 2.0 platforms, where producers and consumers of content unify, has drastically changed the information market. On the one hand, the promise of the 'wisdom of the crowd' has inspired successful projects such as Wikipedia, which has become the primary source of crowd-based information in many languages. On the other hand, the decentralized and often unmonitored environment of such projects may make them susceptible to low-quality content. In this work, we focus on Urban Dictionary, a crowd-sourced online dictionary. We combine computational methods with qualitative annotation and shed light on the overall features of Urban Dictionary in terms of growth, coverage and types of content. We measure a high presence of opinion-focused entries, as opposed to the meaning-focused entries that we expect from traditional dictionaries. Furthermore, Urban Dictionary covers many informal, unfamiliar words as well as proper nouns. Urban Dictionary also contains offensive content, but highly offensive content tends to receive lower scores through the dictionary's voting system. The low threshold to include new material in Urban Dictionary enables quick recording of new words and new meanings, but the resulting heterogeneous content can pose challenges in using Urban Dictionary as a source to study language innovation.


Methodological challenges of genome-wide association analysis in Africa.
Medical research in Africa has yet to benefit from the advent of genome-wide association (GWA) analysis, partly because the genotyping tools and statistical methods that have been developed for European and Asian populations struggle to deal with the high levels of genome diversity and population structure in Africa. However, the haplotypic diversity of African populations might help to overcome one of the major roadblocks in GWA research, the fine mapping of causal variants. We review the methodological challenges and consider how GWA studies in Africa will be transformed by new approaches in statistical imputation and large-scale genome sequencing.


Aberrant sensory innervation of the olfactory bulb in neuropilin-2 mutant mice.
The mammalian olfactory system consists of two anatomically segregated structures, the main olfactory system and the vomeronasal system, which each detect distinct types of chemical stimuli in the environment. During development, sensory neurons establish precise axonal connections with their respective targets within the olfactory bulb. The specificity of the odorant or vomeronasal receptor expressed by the sensory neuron is crucial in this process, yet it is less clear which of the more conventional axon guidance molecules are involved. Here, we show that neuropilin-2, a coreceptor for some of the class 3 semaphorins, is expressed in subpopulations of olfactory and vomeronasal sensory neurons. We generated a knock-out mutation in the neuropilin-2 gene by gene targeting in embryonic stem cells. Neuropilin-2 mutant mice exhibit profound and distinct effects on target innervation within the olfactory bulb. In the main olfactory system, axons of olfactory sensory neurons penetrate into the deeper layers of the main olfactory bulb. In the vomeronasal system, axonal fasciculation within the vomeronasal nerve is affected; some axons are misrouted and innervate glomeruli in an ectopic domain of the accessory olfactory bulb.


Mobilization of endothelial progenitor cells into the circulation in burned patients.
Bone marrow-derived endothelial progenitor cells (EPCs) have been detected in the peripheral blood of patients following thermal injury. EPCs migrate to sites of active neovascularization in response to mediators released after trauma, contributing to wound healing. The aim was to characterize levels and kinetics of EPCs in burned patients, then relate these to key mobilizing factors, vascular endothelial growth factor (VEGF) and the chemokine (C-X-C motif) ligand 12 (CXCL 12), and compare them with those in healthy subjects. The study included 19 adult patients with superficial or full-thickness burns and 50 blood donor volunteer controls. EPCs, identified by cell surface markers CD45(dim/-), CD133+, CD144+ and VEGF receptor 2, were quantified by four-colour flow cytometry. Plasma VEGF and CXCL12 were measured using enzyme-linked immunosorbent assay. Burned patients showed a rapid rise in EPC levels within 24 h, a ninefold increase compared with controls, returning to basal levels by 72 h. Body surface area burned correlated strongly with the degree of mobilization. EPC levels correlated significantly with rises in plasma VEGF and CXCL12. Thermal injury induced a rapid rise in EPCs that was proportional to the extent of the burn and significantly correlated with levels of angiogenic cytokines. Such cytokines may be used to stimulate EPCs as a future therapeutic target in burned patients.


Characterization and risk of exposure to elements from artisanal gold mining operations in the Bolivian Andes.
Artisanal and small-scale gold mining (ASGM) offers low-skilled workers an opportunity to elevate themselves out of poverty. However, this industry operates with little to no pollution controls and the cost to the environment and human health can be large. The objectives of this study were to measure levels of arsenic (As), manganese (Mn), cobalt (Co), lead (Pb), and mercury (Hg) in the environment and characterize health risks to miners and residents in an area with active ASGM operations. An exposure assessment was conducted at two different mining sites and a nearby village in the Bolivian Anders. The resulting measurements were then used to quantify cancerous and noncancerous health risks to children and adults working at and living near ASGM areas. Soil concentrations of As were well above background levels and showed great variations between the village and mining area. Mercury vapor levels at the two mining sites were approximately 30 times larger than the EPA reference concentration. The risk of developing non-cancerous health effects were primarily due to exposure to As and Hg. The probability of individuals developing cancer was considerably increased with adult miners having a probability of 1.3 out of 100. Cancer potential was driven by exposure to As, with de minimus cancer risk from all other elements. Based on the environmental characterization of elements in soils and Hg vapors, the risk of developing cancerous and non-cancerous health outcomes were above a level of concern based on EPA risk assessment guidance. Personal protective equipment was not worn by workers and Hg amalgam is commonly heated in workers' homes. Better education of the risks of ASGM is needed as well as simple controls to reduce exposure.


An aberrant transcription factor network essential for Wnt signaling and stem cell maintenance in glioblastoma.
Glioblastoma (GBM) is thought to be driven by a subpopulation of cancer stem cells (CSCs) that self-renew and recapitulate tumor heterogeneity yet remain poorly understood. Here, we present a comparative analysis of chromatin state in GBM CSCs that reveals widespread activation of genes normally held in check by Polycomb repressors. These activated targets include a large set of developmental transcription factors (TFs) whose coordinated activation is unique to the CSCs. We demonstrate that a critical factor in the set, ASCL1, activates Wnt signaling by repressing the negative regulator DKK1. We show that ASCL1 is essential for the maintenance and in vivo tumorigenicity of GBM CSCs. Genome-wide binding profiles for ASCL1 and the Wnt effector LEF-1 provide mechanistic insight and suggest widespread interactions between the TF module and the signaling pathway. Our findings demonstrate regulatory connections among ASCL1, Wnt signaling, and collaborating TFs that are essential for the maintenance and tumorigenicity of GBM CSCs.


Sexual dysfunction in female partners of men who have undergone radical prostatectomy correlates with sexual dysfunction of the male partner.
Preservation and restoration of erectile function after radical prostatectomy (RP) for prostate cancer has been extensively studied. However, the influence of RP on the sexual function of female partners is poorly understood. The purpose of this retrospective study is to assess sexuality in men who have undergone RP for prostate cancer and their female partners. Men who underwent RP for localized prostate cancer at our institution from 1996 to 2000 were identified and invited to participate in this study with their female partners. Both partners completed a demographic survey. Men completed the International Index of Erectile Function (IIEF) and female partners completed the Female Sexual Function Index (FSFI) and supplemental questions. Correlation between IIEF and FSFI domain scores was determined in matched couples using Pearson correlation coefficient. Kappa statistics and Spearman correlation coefficient were calculated for supplemental questions and IIEF domain scores. Of 1,134 men contacted by letter, 90 (8%) couples completed demographic surveys and both the IIEF/FSFI. Pearson correlation coefficients of IIEF and FSFI domain scores in matched couples demonstrated significant correlation (P<0.05) of all IIEF domains with all FSFI domains with the exception of male erectile function and overall sexual function with female sexual desire. There was moderate agreement between partner supplemental questions and IIEF domain scores. Response rate was very low. FSFI domain scores correlate with IIEF domain scores, indicating an interrelationship between male and female sexual dysfunction in these couples. Evaluation and treatment of sexual dysfunction after RP should involve both partners.


Self-identification as a moderator of the relationship between gambling-related perceived norms and gambling behavior.
This research was designed to evaluate social influences and perceived social norms on gambling behavior among undergraduate students. Furthermore, this research was designed to replicate and extend previous research demonstrating that young adults overestimate the prevalence of gambling among peers, and that the magnitude of overestimation is positively associated with own use (Larimer and Neighbors, Psychol Addict Behav 17:235-243, 2003). We expected that; (1) gambling college students would identify more strongly with other gambling students compared to other students in general; (2) identification with other gambling students would predict gambling behaviors over and above perceived prevalence of gambling; and (3) identification with other gambling students would moderate the association between perceived social norms and gambling behavior. Participants included 1,486 undergraduate students who completed measures assessing gambling quantity and frequency, gambling-related perceived descriptive norms, and identification with groups. Results revealed that perceived norms for gambling were associated with gambling and revealed that students identified more strongly with other students than either gamblers or student gamblers. However, gambling behavior was more strongly associated with identification with gambling students than students in general. There was consistent support for the perspective that social identity moderates the association between perceived norms for gambling and gambling behavior. This research builds on previous examinations of social influences related to gambling and suggests that it may be important to consider the overall prevalence of a given behavior before considering norms-based intervention approaches. Interventions utilizing social norms for gambling may be advised to consider references other than just the typical student.


Phenotypic variability, neurological outcome and genetics background of 6-pyruvoyl-tetrahydropterin synthase deficiency.
This study aimed to investigate the clinical variability and factors implied in the outcome of 6-pyruvoyl-tetrahydropterin synthase deficiency (PTPSd). Biochemical and clinical phenotype, treatment variables, and 6-pyruvoyl-tetrahydropterin synthase (PTS) genotype, were explored retrospectively in 19 Italian patients (12 males and 7 females, aged 4 months to 33 years). According to the level of biogenic amines in cerebrospinal fluid (CSF) at the diagnosis, the patients were classified as mild (6) (normal level) or severe (13) (abnormal low level) form (MF and SF, respectively). Blood Phe ranged from 151 to 1053 micromol/l in MF (mean +/- SD: 698 +/- 403) and 342-2120 micromol/l in SF (mean +/- SD: 1175 +/- 517) (p = 0.063). Patients with MF showed a normal neurological development (a transient dystonia was detected in one), while all SF patients except one presented with severe neurological impairment and only four had a normal neurological development. The outcome of the SF was influenced by the precocity of the treatment. Serial CSF examinations revealed a decline of 5-hydroxyindolacetic acid in MFs and an incomplete restoration of neurotransmitters in SFs: neither obviously affected the prognosis. PTS gene analysis detected 17 different mutations (seven so far unreported) (only one affected allele was identified in three subjects). A good correlation was found between genotype and clinical and biochemical phenotype. The occurrence of brain neurotransmitter deficiency and its early correction (by the therapy) are the main prognostic factors in PTPSd.


Hydrogen-free catalytic fractionation of woody biomass.
The pulping industry could become a biorefinery if the lignin and hemicellulose components of the lignocellulose are valorized. Conversion of lignin into well-defined aromatic chemicals is still a major challenge. Lignin depolymerization reactions often occur in parallel with irreversible condensation reactions of the formed fragments. Here, we describe a strategy that markedly suppresses the undesired condensation pathways and allows to selectively transform lignin into a few aromatic compounds. Notably, applying this strategy to woody biomass at organosolv pulping conditions, the hemicellulose, cellulose, and lignin were separated and in parallel the lignin was transformed into aromatic monomers. In addition, we were able to utilize a part of the lignocellulose as an internal source of hydrogen for the reductive lignin transformations. We hope that the presented methodology will inspire researchers in the field of lignin valorization as well as pulp producers to develop more efficient biomass fractionation processes in the future.


Debriefing 101: training faculty to promote learning in simulation-based training.
Debriefing is recognized as essential for successful simulation-based training. Unfortunately, its effective use is variable. We developed a train the trainer workshop to teach key evidence-based components of effective debriefing. A workshop focusing on best practices for debriefing in surgical simulation-based training was developed for the 2012 Annual Meeting of the Association for Surgical Education. Content emphasized key theoretical concepts related to and evidence-based components of an effective debriefing. Additionally, the workshop incorporated experiential learning via active debriefing following a simulated scenario. Content of the workshop emphasized effective debriefing as the key to learning in simulation-based education. Key elements of debriefing for educators to keep in mind include the following: approach, learning environment, engagement of learners, reaction, reflection, analysis, diagnosis, and application. Effective debriefing is an essential skill for educators involved in surgical simulation-based training. Without it, learning opportunities are missed. Training the trainer in effective debriefing is essential to ensure standardization of practice.


Efficacy of MR vs CT in epilepsy.
We studied 59 seizure patients with CT, MR, and EEG to determine the efficacy of each in the detection of an epileptogenic focus. EEG was most sensitive (67%), MR was next (53%), and CT was least sensitive (42%). MR detected an abnormality in five patients (8%) in whom CT was negative. EEG was positive in each of these patients. CT failed to demonstrate any focal lesion not detected by MR. MR and CT detected focal abnormalities in seven patients (12%) who had negative EEGs. Five of the seven patients had brain tumors. Eighteen of the 26 patients who underwent surgery had positive CT and MR; 14 of these patients had tumors. The remaining eight patients who had surgery all had temporal lobectomies for intractable seizures; none had tumors. In the complex partial seizure subgroup of 34 patients, MR was positive in 44%, CT was positive in 29%, and EEG was positive in 80%. We consider MR to be the imaging procedure of choice for the detection of an epileptogenic focus in seizure patients. When indicated, CT may be performed as a second procedure to try to distinguish neoplasm from thrombosed vascular malformations and other lesions.


Adherence to Lamivudine after an early withdrawal of hepatitis B immune globulin plays an important role in the long-term prevention of hepatitis B virus recurrence.
Lamivudine combined with hepatitis B immune globulin (HBIg) is the standard of care for preventing the recurrence hepatitis B virus after liver transplant. To determine the risk of hepatitis B virus (HBV) recurrence after early withdrawal of HBIg in patients receiving lamivudine maintenance therapy, 20 patients receiving a course of HBIg and lamivudine after transplantation and long-term maintenance therapy with lamivudine and 9 patients receiving HBIg and lamivudine indefinitely were analyzed. The survival rate was 90% after a mean follow-up of 83 months. The HBV recurrence rate was 14% with a mean period of 91 months free from HBV recurrence. Both groups had similar HBV recurrence rates, 15% for the combination and 11% for lamivudine alone. Four patients, 3 of whom were noncompliant with therapy, experienced posttransplant HBV recurrence. Patients who adhere to long-term prophylaxis with lamivudine after early withdrawal of HBIg have a low risk of HBV recurrence, similar to those who receive combination prophylaxis.


Complete Genome Sequence of Plesiomonas shigelloides Type Strain NCTC10360.
Plesiomonas shigelloides is a Gram-negative rod within the Enterobacteriaceae family. It is a gastrointestinal pathogen of increasing notoriety, often associated with diarrheal disease. P. shigelloides is waterborne, and infection is often linked to the consumption of seafood. Here, we describe the first complete genome for P. shigelloides type strain NCTC10360.