Homozygosity mapping identifies the Crumbs homologue 1 (Crb1) gene as responsible for a recessive syndrome of retinitis pigmentosa and nanophthalmos.
The association of retinitis pigmentosa (RP) and microphthalmia has been reported in a number of familial and isolated cases. Here, the results of genetic analysis in a familial case of early RP associated with nanophthalmos are described. Two affected sibs were ascertained from an endogamous population in Mexico. A genome-wide linkage analysis was performed by means of an Affymetrix 250K microarray. Five large regions of homozygosity were demonstrated. The largest interval comprised 15.08 Mb at chromosome 1q31-32.1 and contained the Crumbs homologue-1, CRB1, a gene responsible for a number of recessive retinal dystrophies. Nucleotide sequence analysis demonstrated a c.1125C>G transversion in CRB1 exon 5, predicting a novel p.Tyr375X variant. To our knowledge this is the first instance in which a CRB1 mutation has been associated with early RP and nanophthalmos. Our results suggest a role for CRB1 in promoting axial growth of the eye. Clinical analysis of additional subjects with retinal dystrophies due to CRB1 mutations will help to identify if the high hyperopia, a frequently observed trait in these subjects, could be related to decreased eye axial length (nanophthalmos).


In vitro characterization of cadmium transport along the gastro-intestinal tract of freshwater rainbow trout (Oncorhynchus mykiss).
An in vitro gut sac technique was used to examine the mechanism(s) of cadmium (Cd) uptake along the gastro-intestinal tract (GIT) of rainbow trout (Oncorhynchus mykiss). The spatial distribution of Cd between three compartments (mucus-binding, mucosal epithelium, and transport into blood space) was determined using a modified Cortland saline containing 50μM Cd (as CdCl(2)) labeled with (109)Cd radiotracer. Taking into account total surface areas, the order of relative importance for total Cd uptake rate was: posterior intestine>anterior intestine>stomach>mid intestine. Cd transport was not inhibited by experimentally reducing fluid transport rates by manipulation of osmotic gradients using mannitol, but was sensitive to internal luminal pressure changes, suggesting a mechanosensitive pathway. Q(10) values (1, 11, and 19°C) indicated a facilitated transport of Cd in the anterior- and mid-intestine. The effects of 10mM Ca on the kinetics of Cd uptake suggest the presence of a common uptake pathway for Cd and Ca in the stomach, anterior-, and mid-intestine. Further evidence of a shared route of entry was found using three Ca channel blockers, lanthanum, verapamil, and nifedipine: both voltage-insensitive and voltage-sensitive Ca channels appear to be present in either some, or all portions of the GIT. Elevated Fe (500μM), Mg (50mM), and Zn (500μM) showed varying degrees of inhibition of Cd transport depending on the compartment and segment of the GIT. Overall it appears that there are multiple sites, and mechanisms, of Cd uptake along the GIT of rainbow trout.


Monoamine Oxidase B Inhibitors in Parkinson's Disease.
Parkinson's disease (PD) is a neurodegenerative disorder with a prevalence increasing with age. Oxidative stress and glutamate toxicity are involved in its pathomechanism. There are still many unmet needs of PD patients, including the alleviation of motor fluctuations and dyskinesias, and the development of therapies with neuroprotective potential. To give an overview of the pharmacological properties, the efficacy and safety of the monoamine oxidase B (MAO-B) inhibitors in the treatment of PD, with special focus on the results of randomized clinical trials. A literature search was conducted in PubMed for 'PD treatment', 'MAO-B inhibitors', 'selegiline', 'rasagiline', 'safinamide' and 'clinical trials' with 'MAO-B inhibitors' in 'Parkinson' disease'. MAO-B inhibitors have a favorable pharmacokinetic profile, improve the dopamine deficient state and may have neuroprotective properties. Safinamide exhibits an anti-glutamatergic effect as well. When applied as monotherapy, MAO-B inhibitors provide a modest, but significant improvement of motor function and delay the need for levodopa. Rasagiline and safinamide were proven safe and effective when added to a dopamine agonist in early PD. As add-on to levodopa, MAO-B inhibitors significantly reduced off-time and were comparable in efficacy to COMT inhibitors. Improvements were achieved as regards certain non-motor symptoms as well. Due to the efficacy shown in clinical trials and their favorable side-effect profile, MAO-B inhibitors are valuable drugs in the treatment of PD. They are recommended as monotherapy in the early stages of the disease and as add-on therapy to levodopa in advanced PD.


Virus in a parasitoid wasp: suppression of the cellular immune response in the parasitoid's host.
A virus that replicates in the ovary of a parasitoid wasp is injected into the parasitoid's host during oviposition. Successful development of th parasitoid egg within the host depends on the presence of th virus, which acts to suppress the host's immune response (encapsulation) toward the egg. This is an example of obligatory mutualism between a virus and a eukaryotic organism.


Investigation of the Common Independent Component Analysis Approaches in Biological Signals for Removing Cardiac Field Artefact from EEG.
Electroencephalography (EEG) signals are often impacted by the cardiac field artefact (CFA), which can compromise EEG analysis. Independent component analysis (ICA) has proven effective in removing such artefacts, including CFA. This paper examines three well-known ICA algorithms commonly utilized in EEG signal processing and assesses their ability to decompose EEG into independent components (ICs) to remove CFA. The paper also investigates whether a new two-level ICA approach can improve performance. Results are evaluated using a synthetic dataset of 10 subjects.


Functional neuronal network activity differs with cognitive dysfunction in childhood-onset systemic lupus erythematosus.
Neuropsychiatric manifestations are common in childhood-onset systemic lupus erythematosus (cSLE) and often include neurocognitive dysfunction (NCD). Functional magnetic resonance imaging (fMRI) can measure brain activation during tasks that invoke domains of cognitive function impaired by cSLE. This study investigates specific changes in brain function attributable to NCD in cSLE that have potential to serve as imaging biomarkers. Formal neuropsychological testing was done to measure cognitive ability and to identify NCD. Participants performed fMRI tasks probing three cognitive domains impacted by cSLE: visuoconstructional ability (VCA), working memory, and attention. Imaging data, collected on 3-Tesla scanners, included a high-resolution T1-weighted anatomic reference image followed by a T2*-weighted whole-brain echo planar image series for each fMRI task. Brain activation using blood oxygenation level-dependent contrast was compared between cSLE patients with NCD (NCD-group, n = 7) vs. without NCD (noNCD-group, n = 14) using voxel-wise and region of interest-based analyses. The relationship of brain activation during fMRI tasks and performance in formal neuropsychological testing was assessed. Greater brain activation was observed in the noNCD-group vs. NCD-group during VCA and working memory fMRI tasks. Conversely, compared to the noNCD-group, the NCD-group showed more brain activation during the attention fMRI task. In region of interest analysis, brain activity during VCA and working memory fMRI tasks was positively associated with the participants' neuropsychological test performance. In contrast, brain activation during the attention fMRI task was negatively correlated with neuropsychological test performance. While the NCD group performed worse than the noNCD group during VCA and working memory tasks, the attention task was performed equally well by both groups. NCD in patients with cSLE is characterized by differential activation of functional neuronal networks during fMRI tasks probing working memory, VCA, and attention. Results suggest a compensatory mechanism allows maintenance of attentional performance under NCD. This mechanism appears to break down for the VCA and working memory challenges presented in this study. The observation that neuronal network activation is related to the formal neuropsychological testing performance makes fMRI a candidate imaging biomarker for cSLE-associated NCD.


Th17 cytokines in recall responses against Francisella tularensis in humans.
To determine whether cytokines and T-cell subsets other than Th1 cells contribute to secondary immune responses against Francisella species, we investigated production of Th17-associated cytokines IL-17 and IL-22 in a recall response to Francisella tularensis. Peripheral blood mononuclear cells (PBMCs) from volunteers previously immunized with the F. tularensis live vaccine strain (LVS) were stimulated in vitro with bacterial lysates of LVS or a nonpathogenic type A B38 strain. Gene expression analysis by real-time PCR showed that IL-17 and IL-22 transcripts were induced in immune PBMCs at a significantly higher level than in cells from nonvaccinated volunteers stimulated with LVS or B38 antigens at 24 h. In addition, we detected both cell-associated and secreted IL-22 at 24 h after stimulation and IL-17 at 72 h post-stimulation. Intracellular IL-22 and IL-17 were observed in memory CD4+ cells and less in memory CD8+ cells. These findings suggest that Th17 responses in addition to the Th1 response may play an important role in adaptive immunity against Francisella.


Preliminary evidence of reduced brain network activation in patients with post-traumatic migraine following concussion.
Post-traumatic migraine (PTM) (i.e., headache, nausea, light and/or noise sensitivity) is an emerging risk factor for prolonged recovery following concussion. Concussions and migraine share similar pathophysiology characterized by specific ionic imbalances in the brain. Given these similarities, patients with PTM following concussion may exhibit distinct electrophysiological patterns, although researchers have yet to examine the electrophysiological brain activation in patients with PTM following concussion. A novel approach that may help differentiate brain activation in patients with and without PTM is brain network activation (BNA) analysis. BNA involves an algorithmic analysis applied to multichannel EEG-ERP data that provides a network map of cortical activity and quantitative data during specific tasks. A prospective, repeated measures design was used to evaluate BNA (during Go/NoGo task), EEG-ERP, cognitive performance, and concussion related symptoms at 1, 2, 3, and 4 weeks post-injury intervals among athletes with a medically diagnosed concussion with PTM (n = 15) and without (NO-PTM) (n = 22); and age, sex, and concussion history matched controls without concussion (CONTROL) (n = 20). Participants with PTM had significantly reduced BNA compared to NO-PTM and CONTROLS for Go and NoGo components at 3 weeks and for NoGo component at 4 weeks post-injury. The PTM group also demonstrated a more prominent deviation of network activity compared to the other two groups over a longer period of time. The composite BNA algorithm may be a more sensitive measure of electrophysiological change in the brain that can augment established cognitive assessment tools for detecting impairment in individuals with PTM.


[A population-based study on factors associated with functional disability among older adults in the Great Metropolitan Belo Horizonte, Minas Gerais State, Brazil].
This study assessed factors associated with functional disability in old age in Greater Metropolitan Belo Horizonte, Minas Gerais State, Brazil. The theoretical model was based on predisposing (socio-demographic), extra-individual (social support, use of health services), and intra-individual factors (health conditions). The study was conducted in a probabilistic sample of 1,786 subjects aged > 60 years. The dependent variable was disability defined as mild (some difficulty) or severe (total dependence) in performing at least one basic activity of daily living. Prevalence of disability was 16% (8% mild; 8% severe). Age and worse self-rated health were independently and positively associated with mild and severe disability. Self-reported hypertension and arthritis were associated with mild disability, while diabetes and stroke were associated with severe disability. Severe disability was independently and negatively associated with number of visits by friends in the previous 30 days. According to the results, chronic conditions associated with disability in the study population are preventable, and severely disabled elderly had less extra-family social support.


Novel strains of Culex flavivirus and Hubei chryso-like virus 1 from the Anopheles mosquito in western Kenya.
Surveillance of mosquito vectors is critical for early detection, prevention and control of vector borne diseases. In this study we used advanced molecular tools, such as DNA barcoding in combination with novel sequencing technologies to discover new and already known viruses in genetically identified mosquito species. Mosquitoes were captured using BG sentinel traps in Western Kenya during May and July 2019, and homogenized individually before pooled into groups of ten mosquitoes. The pools and individual samples were then used for molecular analysis and to infect cell cultures. Of a total of fifty-four (54) 10-pools, thirteen (13) showed cytopathic effect (CPE) on VeroB4 cells, eighteen (18) showed CPE on C6/36 cells. Eight (8) 10-pools out of the 31 CPE positive pools showed CPE on both VeroB4 and C6/36 cells. When using reverse transcriptase polymerase chain reaction (RT-PCR), Sanger sequencing and Twist Comprehensive Viral Research Panel (CVRP) (Twist Biosciences), all pools were found negative by RT-PCR when using genus specific primers targeting alphaviruses, orthobunyaviruses and virus specific primers towards o'nyong-nyong virus, chikungunya virus and Sindbis virus (previously reported to circulate in the region). Interestingly, five pools were RT-PCR positive for flavivirus. Two of the RT-PCR positive pools showed CPE on both VeroB4 and C6/36 cells, two pools showed CPE on C6/36 cells alone and one pool on VeroB4 cells only. Fifty individual mosquito homogenates from the five RT-PCR positive 10-pools were analyzed further for flavivirus RNA. Of these, 19 out of the 50 individual mosquito homogenates indicated the presence of flavivirus RNA. Barcoding of the flavivirus positive mosquitoes revealed the mosquito species as Aedes aegypti (1), Mansonia uniformis (6), Anopheles spp (3), Culex pipiens (5), Culex spp (1), Coquilletidia metallica (2) and Culex quinquefasciatus (1). Of the 19 flavivirus positive individual mosquitoes, five (5) virus positive homogenates were sequenced. Genome sequences of two viruses were completed. One was identified as the single-stranded RNA Culex flavivirus and the other as the double-stranded RNA Hubei chryso-like virus 1. Both viruses were found in the same Anopheles spp. homogenate extracted from a sample that showed CPE on both VeroB4 and C6/36 cells. The detection of both viruses in a single mosquito homogenate indicated coinfection. Phylogenetic analyses suggested that the Culex flavivirus sequence detected was closely related to a Culex flavivirus isolated from Uganda in 2008. All four Hubei chryso-like virus 1 segments clusters closely to Hubei chryso-like virus 1 strains isolated in Australia, China and USA. Two novel strains of insect-specific viruses in Anopheles mosquitoes were detected and characterized.


NTBI unveiled by chelatable fluorescent beads.
Non-transferrin-bound iron (NTBI) emerges in plasma of patients with systemic iron overload, but has also been documented in further pathological conditions. Quantification of NTBI can be useful for diagnosis and management of these disorders. However, currently available detection methods are tedious and often inaccurate, hampering wide applicability. In this issue of the Biochemical Journal, Ma et al. report the development of a novel assay for NTBI measurement, based on an iron-sensitive fluorescent probe that is linked to magnetic beads. The approach offers several advantages over existing technology and may bring NTBI assessment closer to the clinic.


t-complex-associated embryonic surface antigen homologous to mLAMP-1. I. Biochemical and molecular analyses.
Previously we described an embryonic cell surface glycoprotein, ESGp, associated with the t-embryonic lethal alleles of the mouse t complex. This antigen is expressed on the cell surface of both early mouse embryos and embryonal carcinoma (EC) cell lines. The antigen is localized to areas of cell-cell contact in EC lines and redistributes to the outer edges of the blastomeres during compaction, thereby indicating a potential role in embryonic cell-cell interaction. We now report that this t-complex-associated ESGp is homologous to the mouse lysosomal-associated membrane protein-1 (LAMP-1). Limited protein sequence analyses of the amino terminal and an internal peptide indicate considerable homology with the LAMP-1 protein. Biochemical parameters such as protein core size, sulfation and phosphorylation status, and resistance to proteolysis also demonstrate homology. While we detect only a single message with a mouse LAMP-1 cDNA probe via Northern blotting, Southern analyses indicate the existence of at least two homologous LAMP-1 genes. Additionally, we present evidence suggesting that ESGp/LAMP-1 serves as a substrate which may be differentially glycosylated by the activities of the gene products of the different t-lethal alleles.


Whole-Genome Sequencing of a Healthy Aging Cohort.
Studies of long-lived individuals have revealed few genetic mechanisms for protection against age-associated disease. Therefore, we pursued genome sequencing of a related phenotype-healthy aging-to understand the genetics of disease-free aging without medical intervention. In contrast with studies of exceptional longevity, usually focused on centenarians, healthy aging is not associated with known longevity variants, but is associated with reduced genetic susceptibility to Alzheimer and coronary artery disease. Additionally, healthy aging is not associated with a decreased rate of rare pathogenic variants, potentially indicating the presence of disease-resistance factors. In keeping with this possibility, we identify suggestive common and rare variant genetic associations implying that protection against cognitive decline is a genetic component of healthy aging. These findings, based on a relatively small cohort, require independent replication. Overall, our results suggest healthy aging is an overlapping but distinct phenotype from exceptional longevity that may be enriched with disease-protective genetic factors. VIDEO ABSTRACT.


Circadian and glucocorticoid regulation of Rev-erbalpha expression in liver.
Rev-erbalpha [NR1D1], a member of the nuclear receptor superfamily, is an orphan receptor that constitutively represses gene transcription. Rev-erbalpha has been shown to play a role in myocyte differentiation and to be induced during adipogenesis. Furthermore, Rev-erbalpha is a regulator of lipoprotein metabolism. It was recently shown that Rev-erbalpha messenger RNA (mRNA) levels oscillate diurnally in rat liver. Here, we report that the circadian rhythm of Rev-erbalpha in liver is maintained in primary cultures of rat hepatocytes. Because glucocorticoids have been shown to regulate other transcription factors with circadian expression, it was furthermore examined whether hepatic Rev-erbalpha expression is also regulated by glucocorticoids. Treatment of rats with dexamethasone resulted in a decrease of Rev-erbalpha mRNA levels by 70% after 6 h. Furthermore, dexamethasone decreased Rev-erbalpha expression in rat primary hepatocytes in a dose-dependent fashion. This effect was mediated by the glucocorticoid receptor because simultaneous addition of the glucocorticoid antagonist RU486 prevented the decrease in Rev-erbalpha mRNA levels by dexamethasone. Protein synthesis inhibition with cycloheximide markedly induced Rev-erbalpha mRNA levels; however, this induction was reduced by dexamethasone supplementation in both rat and human primary hepatocytes. Treatment with actinomycin D blocked the repression of Rev-erbalpha expression by dexamethasone in rat hepatocytes, suggesting that glucocorticoids regulate Rev-erbalpha expression at the transcriptional level. Transient transfection experiments further indicated that Rev-erbalpha promoter activity is repressed by dexamethasone in the presence of cotransfected glucocorticoid receptor. Taken together, these data demonstrate that Rev-erbalpha expression is under the control of both the circadian clock and glucocorticoids in the liver.


Dapsone and Nitroso Dapsone Activation of Naı̈ve T-Cells from Healthy Donors.
Dapsone (DDS) causes hypersensitivity reactions in 0.5-3.6% of patients. Although clinical diagnosis is indicative of a hypersensitivity reaction, studies have not been performed to define whether dapsone or a metabolite activates specific T-cells. Thus, the aims of this study were to explore the immunogenicity DDS and nitroso DDS (DDS-NO) using peripheral blood mononuclear cells from healthy donors and splenocytes from mice and generate human T-cell clones to characterize mechanisms of T-cell activation. DDS-NO was synthesized from DDS-hydroxylamine and shown to bind to the thiol group of glutathione and human and mouse albumin through sulfonamide and N-hydroxyl sulphonamide adducts. Naïve T-cell priming to DDS and DDS-NO was successful in three human donors. DDS-specific CD4+ T-cell clones were stimulated to proliferate in response to drug via a MHC class II restricted direct binding interaction. Cross reactivity with DDS-NO, DDS-analogues, and sulfonamides was not observed. DDS-NO clones were CD4+ and CD8+, MHC class II and I restricted, respectively, and activated via a pathway dependent on covalent binding and antigen processing. DDS and DDS-NO-specific clones secreted a mixture of Th1 and Th2 cytokines, but not granzyme-B. Splenocytes from mice immunized with DDS-NO were stimulated to proliferate in vitro with the nitroso metabolite, but not DDS. In contrast, immunization with DDS did not activate T-cells. These data show that DDS- and DDS-NO-specific T-cell responses are readily detectable.


Occupational exposure to airborne microorganisms, endotoxins and β-glucans in poultry houses at different stages of the production cycle.
The aim of the presented study was to assess the exposure of poultry workers to airborne microorganisms, endotoxins and β-glucans during different stages of the chicken production cycle in 3 commercially-operated poultry houses. Personal and stationary sampling was carried out to assess exposure to both viable and total microbial aerosols. The stationary measurements of PM10 were performed to establish the level of endotoxins and β-glucans. The concentrations of bacterial and fungal aerosols ranged from 2.5×10(2) CFU/m(3)-2.9×10(6) CFU/m(3), and from 1.8×10(2) CFU/m(3)-1.8×10(5) CFU/m(3), respectively. The number of culturable microorganisms was significantly lower than their total counts, constituting from 0.0004%-6.4% of the total microbial flora. The level of PM10 in poultry facilities did not exceed 4.5 mg/m(3). After the flock entered the clean house, the level of endotoxins and β-glucans increased from below detection limit to 8,364 ng/m(3) and from 0.8 ng/m(3) to 6,886 ng/m(3), respectively. The presented study shows that professional activities in poultry farms are associated with constant exposure to bioaerosol, which may pose a health hazard to workers. It was found that workers' exposure to airborne microorganisms increased with consecutive stages of the chicken production cycle.


[The role of enteropathogenic infection, local immunity, and autosensibilisation in pathogenesis of acute surgical diseases of peritoneal cavity by the example of acute appendicitis].
The complex of interrelated pathogenic factors in development of postoperative suppurative inflammatory complication with abdominal cavity organs lesion was studied on the example of an acute appendicitis. It includes enteropathogenic microorganisms, insufficiency of local immunity, deficiency of immunoglobulins, lysozym. Findings show the possibility in seeking the measures of prevention of emergent state, in particular, an acute appendicitis.


Intravenous Delayed Gadolinium-Enhanced MR Imaging of the Endolymphatic Space: A Methodological Comparative Study.
In-vivo non-invasive verification of endolymphatic hydrops (ELH) by means of intravenous delayed gadolinium (Gd) enhanced magnetic resonance imaging of the inner ear (iMRI) is rapidly developing into a standard clinical tool to investigate peripheral vestibulo-cochlear syndromes. In this context, methodological comparative studies providing standardization and comparability between labs seem even more important, but so far very few are available. One hundred eight participants [75 patients with Meniere's disease (MD; 55.2 ± 14.9 years) and 33 vestibular healthy controls (HC; 46.4 ± 15.6 years)] were examined. The aim was to understand (i) how variations in acquisition protocols influence endolymphatic space (ELS) MR-signals; (ii) how ELS quantification methods correlate to each other or clinical data; and finally, (iii) how ELS extent influences MR-signals. Diagnostics included neuro-otological assessment, video-oculography during caloric stimulation, head-impulse test, audiometry, and iMRI. Data analysis provided semi-quantitative (SQ) visual grading and automatic algorithmic quantitative segmentation of ELS area [2D, mm2] and volume [3D, mm3] using deep learning-based segmentation and volumetric local thresholding. Within the range of 0.1-0.2 mmol/kg Gd dosage and a 4 h ± 30 min time delay, SQ grading and 2D- or 3D-quantifications were independent of signal intensity (SI) and signal-to-noise ratio (SNR; FWE corrected, p < 0.05). The ELS quantification methods used were highly reproducible across raters or thresholds and correlated strongly (0.3-0.8). However, 3D-quantifications showed the least variability. Asymmetry indices and normalized ELH proved the most useful for predicting quantitative clinical data. ELH size influenced SI (cochlear basal turn p < 0.001), but not SNR. SI could not predict the presence of ELH. In conclusion, (1) Gd dosage of 0.1-0.2 mmol/kg after 4 h ± 30 min time delay suffices for ELS quantification. (2) A consensus is needed on a clinical SQ grading classification including a standardized level of evaluation reconstructed to anatomical fixpoints. (3) 3D-quantification methods of the ELS are best suited for correlations with clinical variables and should include both ears and ELS values reported relative or normalized to size. (4) The presence of ELH increases signal intensity in the basal cochlear turn weakly, but cannot predict the presence of ELH.


The anti-obesogenic effects of dietary berry fruits: A review.
The prevalence of obesity in the world is fearsomely climbing, which has brought about heavy threats on human health and economic development. For coping with this problem, researchers have looked at the profound potentials of natural products for resolving obesity because of their high efficiencies and few undesirable outcomes in the recent years. Berry fruits are huge reservoirs of bioactive components, and their anti-obesity potentials are arousing much interests. In this review, the current main strategies to manage obesity were summarized, including inhibiting appetite and lowering the food intake, improving energy expenditure and thermogenesis, suppressing absorption and digestion, reducing lipid synthesis and storage as well as modulating composition of gut microbiota. In addition, this review discussed the potentials of dietary berry fruits (blueberries, cranberries, raspberries, strawberries, mulberries, lingonberries, blackberries, black chokeberries, elderberries, bilberries, grape, blackcurrants, jaboticabas, red bayberries, sea-buckthorns, goldenberries and goji berries) to counteract obesity or obesity-associated complications based on recent animal experiments and human studies. Then, the bioaccessibility of phenolic compounds present in berry fruits was discussed. On the other hand, several challenges including securing effective dosage, further understanding their interaction with human tissues, improving bioavailability and protection of functional ingredients during delivery should be taken into account and conquered in the coming years.


Diagnosis of erythroid dysplasia by flow cytometry: a review.
The diagnosis of myelodysplastic syndrome (MDS) is complex. Flow cytometric analysis of the myelomonocytic compartment can be helpful, but it is highly subjective and reproducibility by non-specialized groups is unclear. Analysis of the erythroid lineage by flow cytometry is emerging as potentially more reproducible and easier to conduct, while keeping a high diagnostic performance. We review the evidence in this area, including 1) the use of well-established markers - CD71 and CD36 - and other less well-established markers and parameters; 2) the use of flow cytometric scores for the erythroid lineage; and 3) additional aspects, including the emergence of computational tools and the roles of flow cytometry beyond diagnosis. Finally, we discuss the limitations with the current evidence, including 1) the impact of the sample processing protocol and reagents on the results, 2) the lack of a standard gating strategy, and 3) conceptualization and design issues in the available publications. We end by offering our recommendations for the current use - and our personal take on the value - of the analysis of erythroid lineage by flow cytometry.


Elective nodal irradiation with simultaneous integrated boost stereotactic body radiotherapy for pancreatic cancer: Analyses of planning feasibility and geometrically driven DVH prediction model.
We evaluate the feasibility of the elective nodal irradiation strategy in stereotactic body radiotherapy (SBRT) for pancreatic cancer. Three simultaneous integrated boost (SIB)-SBRT plans (Boost1, Boost2, and Boost3) were retrospectively generated for each of 20 different patients. Boost1 delivered 33 and 25 Gy to PTV1 and PTV2, respectively. Boost2 delivered 40, 33, and 25 Gy to boostCTV, PTV1, and PTV2, respectively. Boost3 delivered 33 and 25 Gy to PTV1 and PTV3, respectively. PTV1 covered the initial standard SBRT plan (InitPlan) gross tumor volume (GTV). PTV2 covered CTVgeom which was created by a 10-mm expansion (15 mm posterior) of GTV. PTV3 covered CTVprop which included elective nodal regions. The boostCTV included GTV as well as involved vasculature. The planning feasibility in each scenario and dose-volume histograms (DVHs) were analyzed and compared with the InitPlan (delivered 33 Gy only to PTV1) by paired t-test. Next, a novel DVH prediction model was developed and its performance was evaluated according to the prediction accuracy (AC) of planning violations. Then, the model was used to simulate the impacts of GTV-to-organs at risk (OAR) distance and gastrointestinal (GI) OAR volume variations on planning feasibility. Significant dose increases were observed in GI-OARs in SIB-SBRT plans when compared with InitPlan. All dose constraints were met in 63% of cases in InitPlan, Boost1, and Boost2, whereas Boost3 developed DVH violations in all cases. Utilizing previous patient anatomy, the novel DVH prediction model achieved a high AC in the prediction of violations for GI-OARs; the positive predictive value, negative predictive value, and AC were 66%, 90%, and 84%, respectively. Experiments with the model demonstrated that the larger proximity volume of GI-OAR at the shorter distance substantially impacted on planning violations. SIB-SBRT plan with geometrically defined prophylactic areas can be dosimetrically feasible, but including all nodal areas with 25 Gy in five fractions appears to be unrealistic.


Effects of sodium acetylsalicylate on body temperature of monkeys under heat exposure.
Exposure of Taiwan monkeys to 38 degrees C ambient temperature caused: a) raised body temperature, b) restlessness and struggling, c) increased evaporative heat loss and d) increase in tail skin temperature. After the administration of sodium acetylsalicylate, 100 to 250 mg/kg intraperitoneally or 4 to 15 mg intracerebroventricularly (third ventricle) the same heat load caused: a) less increase in body temperature, b) no restlessness or struggling, c) an initially higher rate of evaporative heat loss and d) higher tail skin temperature. Larger doses of acetylsalicylate caused greater antihyperthermic effects. Intracerebroventricular injection was more effective than the intraperitoneal route. These data show that acetylsalicylate improved tolerance of animals to external heat loads. The magnitude of body temperature elevation during heat exposure was reduced in the presence of acetylsalicylate by activation of heat-dissipating mechanisms, presumably through the central nervous system.


Therapeutic strategies in an outbreak scenario to treat the novel coronavirus originating in Wuhan, China.
A novel coronavirus (2019-nCoV) originating in Wuhan, China presents a potential respiratory viral pandemic to the world population. Current efforts are focused on containment and quarantine of infected individuals. Ultimately, the outbreak could be controlled with a protective vaccine to prevent 2019-nCoV infection. While vaccine research should be pursued intensely, there exists today no therapy to treat 2019-nCoV upon infection, despite an urgent need to find options to help these patients and preclude potential death. Herein, I review the potential options to treat 2019-nCoV in patients, with an emphasis on the necessity for speed and timeliness in developing new and effective therapies in this outbreak. I consider the options of drug repurposing, developing neutralizing monoclonal antibody therapy, and an oligonucleotide strategy targeting the viral RNA genome, emphasizing the promise and pitfalls of these approaches. Finally, I advocate for the fastest strategy to develop a treatment now, which could be resistant to any mutations the virus may have in the future. The proposal is a biologic that blocks 2019-nCoV entry using a soluble version of the viral receptor, angiotensin-converting enzyme 2 (ACE2), fused to an immunoglobulin Fc domain, providing a neutralizing antibody with maximal breath to avoid any viral escape, while also helping to recruit the immune system to build lasting immunity. The sequence of the ACE2-Fc protein is provided to investigators, allowing its possible use in recombinant protein expression systems to start producing drug today to treat patients under compassionate use, while formal clinical trials are later undertaken. Such a treatment could help infected patients before a protective vaccine is developed and widely available in the coming months to year(s).


Effect of hypoxia and hypercapnia on neurohypophyseal blood flow.
Neurohypophyseal blood flow responses to hypoxia and hypercapnia were studied in pentobarbital anesthetized, paralyzed dogs. Arterial O2 content was lowered from control (18 +/- 2 vol%) to 8 +/- 1 vol% by either decreasing O2 tension (hypoxic hypoxia, HH) or by increasing carboxyhemoglobin saturation (carbon monoxide hypoxia, COH) at normal O2 tension. In all animals HH and COH resulted in similar increases in total cerebral blood flow (239 and 300%, respectively). Regional cerebral blood flow showed a similar increase for all brain regions except the neurohypophysis (NH). The NH increased its blood flow with HH (approximately 320% of control) but was unchanged with COH (117% of control). The responsiveness of NH blood vessels was tested under conditions of hypercapnia (10% CO2) and HH with blood pressure controlled by concurrent hemorrhage. The response of NH vessels to altered arterial O2 tension occurs independently of blood pressure. Systemic [H+] or CO2 tension produce only small changes in NH blood flow. These data suggest that hypoxic and hypercapnic regulatory mechanisms for the NH are different from those of other brain regions. The precise mechanism by which the NH hypoxic response occurs remains unclear, but our data suggest an important role for systemic arterial O2 tension and chemoreceptors.


Modifications and evaluation of smell and taste functions in total laryngectomy: systematic review.
The aim of this study was to systematically review the alterations in smell and taste functions in individuals who underwent total laryngectomy, as well as to identify in the literature some aspects involved in the evaluation of these functions in this population. We performed a systematic review on the databases Medline, LILACS and SciELO, and this data search occurred in October/2010. The search used keywords and free terms, and retrieved 84 articles, 79 from Medline, via Pubmed, and five form LILACS and SciELO. Sixteen of these articles were selected. Most studies in this review attested decline in sensory of smell and taste functions in individuals who underwent total laryngectomy and, although there is consensus regarding the damage caused to these functions by the nasal airflow interruption, it is not yet possible to evidence details in the mechanisms and structures involved, especially regarding the modifications in the olfactory mucous.


Polysialic acid and N-CAM localisation in embryonic rat kidney: mesenchymal and epithelial elements show different patterns of expression.
The expression of polysialic acid (PSA) and the neural cell adhesion molecule (N-CAM) during the embryonic development of rat kidney was investigated using immunocytochemistry and immunoblotting. A monoclonal antibody (mAb 735), which recognised only long chain PSA, and polyclonal antibodies specific for N-CAM were employed. At the earliest stages of metanephros formation, PSA and N-CAM immunostaining was found in both embryonic anlagen, namely the uretic bud and the metanephrogenic mesenchyme. Reactivity in uretic bud derivatives declined during embryonic development and was generally absent in the collecting duct system by embryonic day 18 (E18). Uninduced metanephrogenic mesenchyme was immunostained throughout development while induced mesenchymal cells showed greatly increased PSA and N-CAM immunoreactivity during their transformation into epithelium. This staining declined rapidly as nephrons differentiated. These processes were preceded by sorting of PSA and N-CAM to the basolateral plasma membrane. Similar N-CAM and PSA patterns were observed in mesonephros development. In adult kidney parenchyma both PSA and N-CAM were undetectable. Immunoblotting of samples of embryonic kidney with mAb 735 revealed a broad band ranging from 140 to greater than 200 x 10(3) Mr. N-CAM antibodies revealed reactivity in a band of 140 x 10(3) Mr after removal of PSA by endoneuraminidase treatment. Expression of N-CAM and PSA in both embryonic anlagen indicates that neither molecular species acts primarily as an inductive signal. These molecules were localised in areas where changes in cell adhesion during organogenesis might be important and thus may be involved in the grouping of developing cells.


Exploring the Therapeutic Affordances of Self-Harm Online Support Communities: An Online Survey of Members.
A growing number of online communities have been established to support those who self-harm. However, little is known about the therapeutic affordances arising from engagement with these communities and resulting outcomes. The aim of this study was to explore the presence of therapeutic affordances as reported by members of self-harm online support communities. In total, 94 respondents (aged 13-63 years, mean=23.5 years; 94% female) completed an online survey exploring their experiences of engaging with a self-harm online support community. Respondents varied in terms of how long they had been accessing an online community, with 22% (21/94) accessing less than 1 year, 39% (37/94) 1 to 2 years, 14% (13/94) 2 to 3 years, and 24.5% (23/94) more than 3 years. Responses were analyzed using deductive thematic analysis. The results of our analysis describe each of the five therapeutic affordances that were present in the data, namely (1) connection, the ability to make contact with others who self-harm for the purposes of mutual support and in so doing reduce feelings of loneliness and isolation; (2) adaptation, that is, how use of online support varies in relation to the personal circumstances of the individual user; (3) exploration, that is, the ability to learn about self-harm and learn about strategies to reduce or stop self-harming behavior; (4) narration, that is, the ability to share experiences, as well as read about the experiences of others; and (5) self-presentation, that is, how and what users present about themselves to others in the online community. Our findings suggest that engagement with self-harm online support communities may confer a range of therapeutic benefits for some users, which may serve to minimize the psychosocial burden of self-harm and promote positive coping strategies. In addition, the online nature of the support available may be helpful to those who are unable to access face-to-face support.


Scanning electron microscopy applied to seed-borne fungi examination.
The aim of this study was to test the standard scanning electron microscopy (SEM) as a potential alternative to study seed-borne fungi in seeds, by two different conditions of blotter test and water restriction treatment. In the blotter test, seeds were subjected to conditions that enabled pathogen growth and expression, whereas the water restriction method consisted in preventing seed germination during the incubation period, resulting in the artificial inoculation of fungi. In the first condition, seeds of common bean (Phaseolus vulgaris L.), maize (Zea mays L.), and cotton (Gossypium hirsutum L.) were submitted to the standard blotter test and then prepared and observed with SEM. In the second condition, seeds of cotton (G. hirsutum), soybean (Glycine max L.), and common bean (P. vulgaris L.) were, respectively, inoculated with Colletotrichum gossypii var. cephalosporioides, Colletotrichum truncatum, and Colletotrichum lindemuthianum by the water restriction technique, followed by preparation and observation with SEM. The standard SEM methodology was adopted to prepare the specimens. Considering the seeds submitted to the blotter test, it was possible to identify Fusarium sp. on maize, C. gossypii var. cephalosporioides, and Fusarium oxysporum on cotton, Aspergillus flavus, Penicillium sp., Rhizopus sp., and Mucor sp. on common bean. Structures of C. gossypii var. cephalosporioides, C. truncatum, and C. lindemuthianum were observed in the surface of inoculated seeds.


Biological Variation of Donor-Derived Cell-Free DNA in Stable Lung Transplant Recipients.
Prior studies demonstrate that donor-derived cell-free DNA (dd-cfDNA) in lung transplant recipients may serve as a marker of allograft injury for detecting allograft rejection and infection. Clinical interpretation of dd-cfDNA requires understanding its biological variation in stable lung transplant patients in order to identify abnormal results suggesting underlying allograft dysfunction. This study establishes the biological variation and reference change values (RCV) of dd-cfDNA in stable lung transplant recipients using an analytically validated assay with an established analytic coefficient of variation (CVA). The AlloSure® assay, a targeted, sequencing-based approach, was used to measure plasma dd-cfDNA in a cohort of lung transplant patients at 4 centers that used dd-cfDNA to monitor for allograft dysfunction in preference to surveillance transbronchial biopsy. Patients with stable allograft function and ≥3 dd-cfDNA samples were included. Intraindividual coefficient of variation (CVI), interindividual CV (CVG), index of individuality (II) and the RCV were calculated. Thirty-five patients with a combined 124 dd-cfDNA samples were included in the final analysis. The median dd-cfDNA was 0.31% (interquartile range 0.18%-0.68%), the 97.5th percentile and 95th percentile were 1.3% and 1.0%, respectively. In 30 stable patients with an average of 3.7 tests, the CVI was 25%, CVG 19%, II 1.33, and RCV 70%. In stable lung transplant patients, fluctuations in dd-cfDNA levels of up to 70% or levels less than 1% are within normal biological variation. With further validation, these thresholds may be incorporated into surveillance monitoring algorithms to identify potentially abnormal results indicating allograft dysfunction.


The F-box gene family is expanded in herbaceous annual plants relative to woody perennial plants.
F-box proteins are generally responsible for substrate recognition in the Skp1-Cullin-F-box complexes that are involved in protein degradation via the ubiquitin-26S proteasome pathway. In plants, F-box genes influence a variety of biological processes, such as leaf senescence, branching, self-incompatibility, and responses to biotic and abiotic stresses. The number of F-box genes in Populus (Populus trichocarpa; approximately 320) is less than half that found in Arabidopsis (Arabidopsis thaliana; approximately 660) or Oryza (Oryza sativa; approximately 680), even though the total number of genes in Populus is equivalent to that in Oryza and 1.5 times that in Arabidopsis. We performed comparative genomics analysis between the woody perennial plant Populus and the herbaceous annual plants Arabidopsis and Oryza in order to explicate the functional implications of this large gene family. Our analyses reveal interspecific differences in genomic distribution, orthologous relationship, intron evolution, protein domain structure, and gene expression. The set of F-box genes shared by these species appear to be involved in core biological processes essential for plant growth and development; lineage-specific differences primarily occurred because of an expansion of the F-box genes via tandem duplications in Arabidopsis and Oryza. The number of F-box genes in the newly sequenced woody species Vitis (Vitis vinifera; 156) and Carica (Carica papaya; 139) is similar to that in Populus, supporting the hypothesis that the F-box gene family is expanded in herbaceous annual plants relative to woody perennial plants. This study provides insights into the relationship between the structure and composition of the F-box gene family in herbaceous and woody species and their associated developmental and physiological features.


Influence of Sodium Periodate and Tyrosinase on Binding of Alginate to Adlayers of Mytilus edulis Foot Protein 1.
Mytilus edulis foot protein 1 (Mefp-1) is the most well-characterized component of this sea mussel's adhesive plaque. The plaque is a condensed, heterogeneous mixture consisting of a large proportion of cross-linked biopolymers that bonds the mussel to a chosen mooring. Mefp-1 is densely populated with lysine and L-3,4-dihyroxyphenylalanine (L-dopa) residues incorporated into a repeating amino acid sequence motif. It has been proposed that one plaque cross-linking reaction is the nucleophilic addition of the epsilon-amino groups of the lysine residues into the oxidized catechol (o-diphenol) functionality (quinone) of the L-dopa residues. In order to determine if this reaction occurs in adlayers of Mefp-1, a previously developed assay for epsilon-amino groups was applied. Adlayers of Mefp-1 were exposed to an oxidant, either the enzyme, mushroom tyrosinase, or sodium periodate. Binding of alginate to adlayers was used to probe for accessibility of epsilon-amino groups. It was found that lysine residues lose the ability to bind alginate after exposure to sodium periodate, but that this loss is not clearly due to a reaction with L-dopa residues. There is a slight decrease of binding of alginate to adlayers of Mefp-1 exposed to either active or thermally deactivated mushroom tyrosinase, probably due to the obstruction of binding sites by bound enzyme. Adsorption kinetics of mushroom tyrosinase onto adlayers of Mefp-1 for active and thermally inactivated enzyme were nearly identical. Attenuated total reflection Fourier transform infrared spectroscopy was used to characterize these interactions at a germanium (Ge) interface. Copyright 2000 Academic Press.


Post-discharge prevention of deep vein thrombosis following total joint replacement.
Recent prospective studies demonstrate that the peak incidence of deep vein thrombosis (DVT) occurs on the fifth postoperative day. If the patient is to be discharged from an acute care facility on day 3 through 6, then the patient will be removed from direct supervision by professionals-frequently observing the patient in an acute care hospital during the high-risk period for the development of DVT or pulmonary embolism. In addition, the period of vulnerability for the development of thromboembolic disease in this group of patients appears to occur for an extended period of time, eg, 3 months. This change in practice has altered the surgeon's approach to the selection of agents for the prevention of thromboembolic complications. Most surgeons wish to administer an agent in the acute care setting that can be continued after the patient is discharged to the rehabilitation facility, subacute care facility, or home. This information reflects the belief of most orthopedic surgeons that prophylactic therapy should be continued beyond the time frame in which the patient is confined to the acute care facility. Although progress has been made in identifying effective prophylactic agents, questions remain as to the appropriate duration of prophylaxis. Based upon available data, it would appear that prophylaxis against thromboembolic disorders should be continued until the patient is spending the majority of his or her day ambulating with or without external aides.


Suitability of [11, 12-3H2]chenodeoxycholic acid and [11, 12-3H2]lithocholic acid for isotope dilution studies of bile acid metabolism in man.
Studies were carried out to assess the stability of the label in [11,12-(3)H(2)]chenodeoxycholic acid and [11,12-(3)H(2)]lithocholic acid during enterohepatic cycling and thus define the suitability of these tracers for isotope dilution studies of bile acids in man. To do this, [11,12-(3)H(2)]-chenodeoxycholic acid and [24-(14)C]chenodeoxycholic acid were administered simultaneously to six healthy adults. The (3)H/(14)C ratio of biliary chenodeoxycholic acid and the specific activity of biliary chenodeoxycholic acid were determined to permit calculation of pool size and turnover rate by the conventional isotope dilution procedure of Lindstedt (1957. Acta Physiol. Scand. 40: 1). Excretion of label in urine was measured as well as the (3)H/(14)C ratio of fecal bile acids. Similar studies were carried out with [11,12-(3)H(2)]lithocholic acid and [24-(14)C]lithocholic acid. With [11,12-(3)H(2)]chenodeoxycholic acid, 10-15% of the label was lost during the first 24 hours, after which the (3)H/(14)C ratio of biliary chenodeoxycholic acid remained constant. The loss of (3)H caused a consistent overestimate of pool size by 10-15%, but since the fractional turnover rate of the two isotopes was identical, the calculated rate of synthesis was 10-15% greater when determined using (3)H. The loss of (3)H was attributed to the presence of (3)H in positions other than 11 and 12 occurring during the labeling procedure when the unsaturated precursor was exposed to carrier-free tritium gas. The (3)H/(14)C ratio of feces was consistently below that of bile, suggesting bacterial removal of additional (3)H during distal intestinal passage. In contrast, [11,12-(3)H(2)]lithocholic acid lost (3)H continuously, so that by 2 days after administration, the (3)H/(14)C ratio had fallen to nearly half of that of the administered mixture. It is concluded that isotope dilution studies may be carried out with one preparation of [11,12-(3)H(2)]chenodeoxycholic acid with an error of 10-15%; possibly a more stable preparation can be prepared with a gentler tritiation procedure. One preparation of [11,12-(3)H(2)]lithocholic acid was unsatisfactory for isotope dilution studies in man.


The association between IgG4 antibodies to dietary factors, islet autoimmunity and type 1 diabetes: the Diabetes Autoimmunity Study in the Young.
Infant dietary exposures have been linked to type 1 diabetes (T1D) development. IgG4 antibody responses to food antigens are associated with food intolerances but have not been explored prospectively in the period preceding T1D. Using a case-cohort design, IgG4 antibodies to ß-lactoglobulin, gluten, and ovalbumin were measured in plasma collected annually from 260 DAISY participants. Of those, 77 developed islet autoimmunity (IA), defined as positive for either insulin, GAD65 or IA-2 autoantibodies on two consecutive visits, and 22 developed T1D. In mixed model analysis adjusting for HLA-DR status, T1D family history, age and ethnicity, higher ß-lactoglobulin IgG4 concentrations were associated with shorter breastfeeding duration (beta = -0.03, 95% Confidence Interval: -0.05, -0.006) and earlier first cow's milk exposure (beta = -0.04, 95% Confidence Interval: -0.08, 0.00). Higher gluten IgG4 was associated with older age at gluten introduction (beta = 0.06, 95% Confidence Interval: 0.00, 0.13). In proportional hazards analysis adjusting for HLA-DR status, T1D family history and ethnicity, IgG4 against individual or multiple dietary antigens throughout childhood were not associated with IA. In addition, mean antigen-specific IgG4 concentrations in infancy (age <2 years) were not associated with risk of IA nor progression to T1D. Higher ovalbumin IgG4 at first IA positive visit was marginally associated with progression to T1D (Hazard Ratio: 1.39, 95% Confidence Interval: 1.00, 1.92). We found no association between the IgG4 response to β-lactoglobulin, gluten, and the development of either IA or T1D. The association between higher ovalbumin and progression to T1D in children with IA should be explored in other populations.


Sequence analysis of temperature-sensitive mutations in the Saccharomyces cerevisiae gene CDC28.
Eleven independently isolated temperature-sensitive mutations in the cell division cycle gene CDC28 were mapped with respect to the DNA sequence of the wild-type gene and then sequenced to determine the precise nature of each mutation. The set yielded six different point mutations, each of which predicts a single amino acid substitution in the CDC28 product. The positions of the mutations did not correlate in any obvious way with observable biological characteristics of the mutant alleles. When the positions of substitutions were collated with a predicted secondary structural analysis of the CDC28 protein kinase, they were found to correlate strongly with probable regions of structural transition.


Who Will Use Pre-Exposure Prophylaxis (PrEP) and Why?: Understanding PrEP Awareness and Acceptability amongst Men Who Have Sex with Men in the UK--A Mixed Methods Study.
Recent clinical trials suggest that pre-exposure prophylaxis (PrEP) may reduce HIV transmission by up to 86% for men who have sex with men (MSM), whilst relatively high levels of PrEP acceptability have been reported to date. This study examines PrEP awareness amongst sub-groups of MSM communities and acceptability amongst MSM in a low prevalence region (Scotland, UK), using a mixed methods design. Quantitative surveys of n = 690 MSM recruited online via social and sociosexual media were analysed using descriptive statistics and multivariate logistic regression. In addition, n = 10 in-depth qualitative interviews with MSM were analysed thematically. Under one third (29.7%) of MSM had heard of PrEP, with awareness related to living in large cities, degree level education, commercial gay scene use and reporting an HIV test in the last year. Just under half of participants (47.8%) were likely to use PrEP if it were available but there was no relationship between PrEP acceptability and previous PrEP awareness. Younger men (18-25 years) and those who report higher risk UAI were significantly more likely to say they would use PrEP. Qualitative data described specific PrEP scenarios, illustrating how risk, patterns of sexual practice and social relationships could affect motivation for and nature of PrEP use. These findings suggest substantial interest PrEP amongst MSM reporting HIV risk behaviours in Scotland. Given the Proud results, there is a strong case to investigate PrEP implementation within the UK. However, it appears that disparities in awareness have already emerged along traditional indicators of inequality. Our research identifies the need for comprehensive support when PrEP is introduced, including a key online component, to ensure equity of awareness across diverse MSM communities (e.g. by geography, education, gay scene use and HIV proximity), as well as to responding to the diverse informational and sexual health needs of all MSM communities.


Implant metastasis of gallbladder carcinoma in situ in a cholecystectomy scar: a case report.
A subcutaneous mass within the scar left by cholecystectomy with common bile duct exploration and T-tube drainage developed 6 years after surgery. Pathologic examination of this mass showed features of atypical villous hyperplasia, similar to that identified within the previously removed gallbladder, but with additional foci of carcinoma in situ. Since excision of the mass, the patient has had persistent fluid collections requiring frequent aspiration. Cytologic analysis of the fluid has revealed tumor cells. The cause of this spread has been unclear. Few literature reports have identified biliary drainage techniques as a source for metastatic seeding. The malignant or metastatic potential of severe dysplasia or carcinoma in situ of the gallbladder associated with T-tube drainage and implantation in the drainage tract is previously unreported.


Two series of new semisynthetic triterpene derivatives: differences in anti-malarial activity, cytotoxicity and mechanism of action.
The discovery and development of anti-malarial compounds of plant origin and semisynthetic derivatives thereof, such as quinine (QN) and chloroquine (CQ), has highlighted the importance of these compounds in the treatment of malaria. Ursolic acid analogues bearing an acetyl group at C-3 have demonstrated significant anti-malarial activity. With this in mind, two new series of betulinic acid (BA) and ursolic acid (UA) derivatives with ester groups at C-3 were synthesized in an attempt to improve anti-malarial activity, reduce cytotoxicity, and search for new targets. In vitro activity against CQ-sensitive Plasmodium falciparum 3D7 and an evaluation of cytotoxicity in a mammalian cell line (HEK293T) are reported. Furthermore, two possible mechanisms of action of anti-malarial compounds have been evaluated: effects on mitochondrial membrane potential (ΔΨm) and inhibition of β-haematin formation. Among the 18 derivatives synthesized, those having shorter side chains were most effective against CQ-sensitive P. falciparum 3D7, and were non-cytotoxic. These derivatives were three to five times more active than BA and UA. A DiOC(6)(3) ΔΨm assay showed that mitochondria are not involved in their mechanism of action. Inhibition of β-haematin formation by the active derivatives was weaker than with CQ. Compounds of the BA series were generally more active against P. falciparum 3D7 than those of the UA series. Three new anti-malarial prototypes were obtained from natural sources through an easy and relatively inexpensive synthesis. They represent an alternative for new lead compounds for anti-malarial chemotherapy.


Pseudoporphyria induced by dialysis treated with oral N-acetylcysteine.
Pseudoporphyria is a rare bullous dermatosis that clinically and histopathologically is similar to porphyria cutanea tarda. It mainly affects patients with chronic renal failure on peritoneal dialysis or hemodialysis. Medications can also be involved in the etiology. Diagnosis and management of this condition is a challenge for dermatologists. The authors report a case of pseudoporphyria related to dialysis with favorable outcome after the use of oral N-acetylcysteine.


Excitability of motor cortex inhibitory circuits in Tourette syndrome before and after single dose nicotine.
The pathophysiology underlying the involuntary tics of Gilles de la Tourette syndrome (GTS) remains unknown. Here we used transcranial magnetic stimulation (TMS) to examine the excitability of two different inhibitory systems in the human motor cortex: short interval intracortical inhibition (SICI) and short interval afferent inhibition (SAI) in 10 healthy non-smoking controls and eight untreated non-smoking patients with GTS. Compared with the healthy control group, both SICI (measured at a range of conditioning intensities) and SAI were reduced in patients. This is consistent with the suggestion that reduced excitability of cortical inhibition is one factor that contributes to the difficulty that patients have in suppressing involuntary tics. In addition, the reduced SAI indicates that impaired intracortical inhibition may not be limited to the motor cortex but also involves circuits linking sensory input and motor output. A single dose of nicotine reduced tic severity as assessed by blind video scoring in the majority of patients. In addition, it abolished the difference between patients and controls in SICI and SAI. There was no effect of nicotine, and no difference between controls and patients in measures of motor or SICI threshold. This indicates that cholinergic input can modulate the efficiency of SICI and SAI differently in GTS and healthy controls.


A JNK1/AP-1-dependent, COX-2 induction is implicated in 12-O-tetradecanoylphorbol-13-acetate-induced cell transformation through regulating cell cycle progression.
Cyclooxygenase-2 (COX-2) is reported to be one of the early-response gene products induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). However, the relevance of COX-2 in TPA-induced cell transformation and the underlying mechanisms remains to be explored. Initially, we verified COX-2 induction after TPA treatment in mouse embryonic fibroblasts (MEF) and mouse epidermal cells Cl 41. More importantly, introduction of COX-2 small interfering RNA in MEFs or Cl 41 cells suppressed the cell transformation caused by TPA treatment. This inhibition could be reversed by overexpression of human full-length COX-2, indicating that COX-2 is at least one of the critical molecules involved in TPA-induced cell transformation. We further showed that TPA-promoted cell cycle progression was partially suppressed by COX-2 small interfering RNA, indicating that COX-2 also participated in TPA-associated cell cycle progression. Investigation of the upstream signaling pathways revealed that c-Jun-NH(2)-kinase 1 (JNK1), but not JNK2, played important roles in COX-2 induction, because knockout of JNK1 gene rather than JNK2 gene markedly impaired COX-2 induction. Furthermore, inhibition of c-Jun/activator protein 1 pathway or JNKs/c-Jun pathway by overexpression of dominant negative mutants of c-Jun, or MKK4 and MKK7 together, resulted in impairment of COX-2 induction, suggesting that JNK1/c-Jun/activator protein 1 pathway is involved in TPA-associated COX-2 induction. In contrast, IKK/p65 nuclear factor-kappaB pathway was not implicated because knockout of IKKalpha, IKKbeta, or p65 gene did not affect COX-2 induction although nuclear factor-kappaB was activated by TPA. In addition, the TPA-promoted cell cycle progression was found impaired in JNK1-deficient, but not in JNK2-deficient, MEFs. Our results show that JNK1-associated COX-2 induction is implicated in TPA-associated cell transformation and cell cycle progression.


Neurofeedback as a Treatment for Impulsivity in a Forensic Psychiatric Population With Substance Use Disorder: Study Protocol of a Randomized Controlled Trial Combined With an N-of-1 Clinical Trial.
Impulsivity and substance use disorder (SUD) are strongly interconnected, with persons scoring high on impulsivity being more vulnerable to develop substance abuse, facing more challenges for successful treatment, and being more prone to engage in criminal behavior. Studies have shown that impulsivity and craving for substances are strongly correlated. Neurofeedback is an effective treatment to reduce impulsive behavior. This study intends to determine to what extent a neurofeedback-intervention that is aimed at reducing impulsivity can also reduce levels of craving in forensic patients with SUD and comorbid Axis I and/or II diagnoses. The main objective of this study is to investigate to what extent a reduction in impulsivity by a sensorimotor rhythm (SMR)-neurofeedback intervention will lead to a reduction in craving in a population of forensic psychiatric patients with a diagnosis of SUD. Participants will be male SUD patients with various comorbidities residing in an inpatient forensic treatment facility approached through treatment supervisors for participation. Participants have tested positive for drug use in the past 24 months. The study consists of 2 parts: a randomized controlled trial (RCT) and a n-of-1 clinical series. In the RCT, 50 patients will be randomly assigned to an intervention (n=25) or a control (n=25) condition. Patients in the intervention group will receive 20 SMR neurofeedback sessions aimed at reducing impulsivity; participants in the control group receive treatment-as-usual (TAU). Additionally, 4 in depth n-of-1 clinical trials will be conducted where effects of an SMR neurofeedback intervention will be compared to effects of sham neurofeedback. Results of this study are expected by the end of 2017. This protocol describes the design of a study testing the effects of an impulsivity-based neurofeedback protocol among forensic patients with SUD and various comorbidities. We expect a significant reduction in impulsive behavior, level of craving, and actual drug-use for participants receiving the SMR neurofeedback protocol. The n-of-1 approach might help to explain effects possibly found in the RCT study since it allows for a more direct focus on treatment effects by following participants more closely and thereby being able to directly attribute behavioral and neurophysiological change to the SMR neurofeedback protocol employed. Dutch National Trial Register NTR5386; http://www.trialregister.nl/trialreg/admin/rctview.asp?TC=5386 (Archived by WebCite at http://www.webcitation.org/6nXLQuoLl).


Growth rate consists of baseline and systematic deviation components in Thoroughbreds.
The objective of this study was to establish a procedure for differentiating a baseline curve from a systematic deviation in weight-age data, and hence to develop a physiological growth model for the Thoroughbred. A total of 2,698 records for 175 foals was obtained during a period of 8 yr (1994 to 2001). Weight-age data were fit with a sigmoid growth equation, W = A(1 + be(-kt))M, where W is BW at age t, A is the asymptotic value of W, b is a scaling parameter that defines the degree of maturity at t = 0, k is a rate constant, and M defines the point of inflection in the sigmoid curve in relation to age. Short-term systematic deviations in the weight-age data were identified by a goodness-of-fit procedure and illustrated in three-dimensional contour plots of the sigmoid equation parameters as they changed upon removal of selected subsets of the data. Based on features of the contour plots, a negative deviation between 210 and 420 d of age was set aside, with the remaining data establishing the baseline data set. The sigmoid growth equation was fit to the baseline data set using a nonlinear mixed model with repeated measures, and indicated a mature weight of 542 +/- 6.2 kg reached at 7 yr. The systematic deviation identified in this weight-age data set is present in other published Thoroughbred growth data and is likely to result in erroneous parameter estimates if not set aside before fitting sigmoid growth equations to the thus-modified weight-age data set. The techniques developed in this study enable identification of short-term systematic deviations in weight-age data and define a realistic baseline growth curve. Differentiation of these two components enables the development of a physiological model of growth that distinguishes between baseline growth and environmental influences, represented respectively, by the baseline curve and the systematic deviation.


Unconditionally stable finite-difference time-domain methods for modeling the Sagnac effect.
We present two unconditionally stable finite-difference time-domain (FDTD) methods for modeling the Sagnac effect in rotating optical microsensors. The methods are based on the implicit Crank-Nicolson scheme, adapted to hold in the rotating system reference frame-the rotating Crank-Nicolson (RCN) methods. The first method (RCN-2) is second order accurate in space whereas the second method (RCN-4) is fourth order accurate. Both methods are second order accurate in time. We show that the RCN-4 scheme is more accurate and has better dispersion isotropy. The numerical results show good correspondence with the expression for the classical Sagnac resonant frequency splitting when using group refractive indices of the resonant modes of a microresonator. Also we show that the numerical results are consistent with the perturbation theory for the rotating degenerate microcavities. We apply our method to simulate the effect of rotation on an entire Coupled Resonator Optical Waveguide (CROW) consisting of a set of coupled microresonators. Preliminary results validate the formation of a rotation-induced gap at the center of a transfer function of a CROW.


SRSF2 Regulation of MDM2 Reveals Splicing as a Therapeutic Vulnerability of the p53 Pathway.
MDM2 is an oncogene and critical negative regulator of tumor suppressor p53. Genotoxic stress causes alternative splicing of MDM2 transcripts, which leads to alterations in p53 activity and contributes to tumorigenesis. MDM2-ALT1 is one of the alternatively spliced transcripts predominantly produced in response to genotoxic stress, and is comprised of terminal coding exons 3 and 12. Previously, we found that SRSF1 induces MDM2-ALT1 by promoting MDM2 exon 11 skipping. Here we report that splicing regulator SRSF2 antagonizes the regulation of SRSF1 by facilitating the inclusion of exon 11 through binding at two conserved exonic splicing enhancers. Overexpression of SRSF2 reduced the generation of MDM2-ALT1 under genotoxic stress, whereas SRSF2 knockdown induced the expression of MDM2-ALT1 in the absence of genotoxic stress. Blocking the exon 11 SRSF2-binding sites using oligonucleotides promoted MDM2-ALT1 splicing and induced p53 protein expression, and apoptosis in p53 wild-type cells. The regulation of MDM2 splicing by SRSF2 is also conserved in mice, as mutation of one SRSF2-binding site in Mdm2 exon 11, using CRISPR-Cas9, increased the expression of the MDM2-ALT1 homolog Mdm2-MS2. IMPLICATIONS: Taken together, the data indicate that modulating MDM2 splicing may be a useful tool for fine-tuning p53 activity in response to genotoxic stress.


Quantitation of myonecrosis induced by myotoxin a from prairie rattlesnake (Crotalus viridis viridis) venom.
Using a pure myotoxin (myotoxin a) isolated from prairie rattlesnake Crotalus viridis viridis) venom, a comparison between two methods for quantitating myonecrosis in mice was made. Measurement of creatine kinase (CK) levels in the plasma showed two peaks in CK levels after myotoxin injection, whereas measurement by histological assay (vacuolation index) showed only one peak. The first peak in CK levels at 3 hr after injection did not correlate with a high vacuolation index, but did correlate with contraction of muscle induced by the toxin. However, there was good correlation between the two methods at 6, 12, 24, 48 and 72 hr after injection, at which time the muscle cells were necrotic. In the case of the first CK peak this method might be measuring contraction of the muscle, but the second peak was probably measuring altered permeability of the sarcolemma since vacuolation, i.e. swelling of sarcoplasmic reticulum, did follow. High plasma CK levels usually preceded high vacuolation indexes, indicating that CK release due to altered sarcolemma permeability preceded vacuolation. Caution should be taken when using plasma CK levels to estimate the quantity of necrotic muscle cells, so as to insure that pathologic and not physiologic changes are being measured.


IL-12 and IL-23 modulate plasticity of FoxP3+ regulatory T cells in human Leprosy.
Leprosy is a bacterial disease caused by M. leprae. Its clinical spectrum reflects the host's immune response to the M. leprae and provide an ideal model to investigate the host pathogen interaction and immunological dysregulation. Tregs are high in leprosy patients and responsible for immune suppression of the host by producing IL-10 and TGF-β cytokines. In leprosy, plasticity of Tregs remain unstudied. This is the first study describing the conversion of Tregs into Th1-like and Th17-like cells using in vitro cytokine therapy in leprosy patients. Peripheral blood mononuclear cells from leprosy patients were isolated and stimulated with M. leprae antigen (MLCwA), rIL-12 and rIL-23 for 48h. Expression of FoxP3 in CD4+CD25+ Tregs, intracellular cytokines IFN-γ, TGF-β, IL-10 and IL-17 in Tregs cells were evaluated by flow cytometry (FACS) after stimulation. rIL-12 treatment increases the levels of pStat4 in Tregs and IFN-γ production. In the presence of rIL-23, pStat3+ and IL-17A+ cells increase. rIL-12 and r-IL-23 treatment downregulated the FoxP3 expression, IL-10 and TGF-β production by Tregs and enhances the expression of co-stimulatory molecules (CD80, CD86). In conclusion rIL-12 converts Tregs into IFN-γ producing cells through STAT-4 signaling while rIL-23 converts Tregs into IL-17 producing cells through STAT-3 signaling in leprosy patients. This study may helpful to provide a new avenue to overcome the immunosuprression in leprosy patients using in vitro cytokine.


Genetically Distinct Behavioral Modules Underlie Natural Variation in Thermal Performance Curves.
Thermal reaction norms pervade organismal traits as stereotyped responses to temperature, a fundamental environmental input into sensory and physiological systems. Locomotory behavior represents an especially plastic read-out of animal response, with its dynamic dependence on environmental stimuli presenting a challenge for analysis and for understanding the genomic architecture of heritable variation. Here we characterize behavioral reaction norms as thermal performance curves for the nematode Caenorhabditis briggsae, using a collection of 23 wild isolate genotypes and 153 recombinant inbred lines to quantify the extent of genetic and plastic variation in locomotory behavior to temperature changes. By reducing the dimensionality of the multivariate phenotypic response with a function-valued trait framework, we identified genetically distinct behavioral modules that contribute to the heritable variation in the emergent overall behavioral thermal performance curve. Quantitative trait locus mapping isolated regions on Chromosome II associated with locomotory activity at benign temperatures and Chromosome V loci related to distinct aspects of sensitivity to high temperatures, with each quantitative trait locus explaining up to 28% of trait variation. These findings highlight how behavioral responses to environmental inputs as thermal reaction norms can evolve through independent changes to genetically distinct modular components of such complex phenotypes.


Gene transfer of hepatocyte growth factor improves angiogenesis and function of chronic ischemic myocardium in canine heart.
Hepatocyte growth factor (HGF) induces angiogenesis in myocardium. In the present study, its effects in chronic ischemic myocardium were tested. Four weeks after left anterior descending coronary artery ligation in canine hearts, HVJ-liposome containing either human HGF gene (160 microg; HGF group, n = 7) or nothing (control group, n = 6) was directly injected into ischemic myocardium. Four weeks after gene transfection, the thickness fraction (TF), an index of regional myocardial contractility (assessed by epicardial pulse-Doppler crystals), the myocardial perfusion flow (assessed by color microspheres), and the capillary density (assessed by immunostaining of vessels) were evaluated in ischemic myocardium. Thickness fraction (percent of nonischemic myocardium) was significantly improved in the HGF group (80 +/- 15 from 52 +/- 16 of pregene; p < 0.05) whereas it was not changed in the control group (52 +/- 10 from 50 +/- 8 of pregene). The perfusion flow (% of nonischemic myocardium) was significantly improved in the HGF group (98 +/- 17 from 51 +/- 14 of pregene; p < 0.05) while it was not changed in the Control group (58 +/- 13 from 62 +/- 18 of pregene). The capillary density was significantly higher in the HGF group (894 +/- 211/mm2; p < 0.05) than that in the control group (511 +/- 127/mm2). Gene transfection of HGF improved angiogenesis, thereby improved regional myocardial function and perfusion in chronic ischemic myocardium. It indicates a potent therapeutic value of HGF gene transfection for chronic ischemic heart diseases such as myocardial infarction.


Roles of the C terminus of Armadillo in Wingless signaling in Drosophila.
Drosophila melanogaster Armadillo and its vertebrate homolog beta-catenin play multiple roles during development. Both are components of cell-cell adherens junctions and both transduce Wingless (Wg)/Wnt intercellular signals. The current model for Wingless signaling proposes that Armadillo binds the DNA-binding protein dTCF, forming a bipartite transcription factor that activates Wingless-responsive genes. In this model, Armadillo's C-terminal domain is proposed to serve an essential role as a transcriptional activation domain. In Xenopus, however, overexpression of C-terminally truncated beta-catenin activates Wnt signaling, suggesting that the C-terminal domain might not be essential. We reexamined the function of Armadillo's C terminus in Wingless signaling. We found that C-terminally truncated mutant Armadillo has a deficit in Wg-signaling activity, even when corrected for reduced protein levels. However, we also found that Armadillo proteins lacking all or part of the C terminus retain some signaling ability if overexpressed, and that mutants lacking different portions of the C-terminal domain differ in their level of signaling ability. Finally, we found that the C terminus plays a role in Armadillo protein stability in response to Wingless signal and that the C-terminal domain can physically interact with the Arm repeat region. These data suggest that the C-terminal domain plays a complex role in Wingless signaling and that Armadillo recruits the transcriptional machinery via multiple contact sites, which act in an additive fashion.


Absorption of eicosapentaenoic acid and docosahexaenoic acid from fish oil triacylglycerols or fish oil ethyl esters co-ingested with a high-fat meal.
The absorption of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil triacylglycerols and fish oil ethyl esters consumed in a high-fat meal (44 g total fat) by male volunteers was measured and compared to values previously reported for consumption in a low-fat meal (8 g total fat). Absorption of EPA, but not of DHA, from fish oil triacylglycerols was significantly improved from 69% to 90% by co-ingestion with the high-fat meal. Absorption of both EPA and DHA from fish oil ethyl esters was increased three-fold, to about 60%, by co-ingestion with the high-fat meal, indicating that absorption of fatty acid ethyl esters is highly dependent on the amount of co-ingested fat.


Main complications of hip arthroplasty: pictorial essay.
Hip arthroplasty is a widely used and successful orthopedic procedure for the treatment of degenerative, inflammatory, or traumatic joint disease. The procedure promotes significant pain relief, as well as recovery of limb function, reduction of disability, and better quality of life. However, there are related complications, which have characteristic imaging aspects. In the present study, we review the literature and exemplify such complications using images obtained at our facility, illustrating the main radiological aspects of complications such as heterotopic ossification, periprosthetic fractures, osteolysis, infection, wear, and dislocation.


Application of "in vivo cryotechnique" to detect erythrocyte oxygen saturation in frozen mouse tissues with confocal Raman cryomicroscopy.
To measure oxygen saturation (SO2) of flowing erythrocytes in blood vessels of living animals, our "in vivo cryotechnique" (IVCT) was combined with confocal Raman microscopy at low temperature (-150 degrees C), referred to as cryomicroscopy. We evaluated two resonance Raman (RR) shifts around 1355 and 1378 cm(-1), reflecting de-oxygenated and oxygenated hemoglobin molecular structures, respectively. Judging from the calibration analyses of quickly frozen human whole blood for the control experiment in vitro, the two RR shifts were well retained at the low temperature, and their calculated ratios mostly reflected the relative SO2 measured with a blood-gas analyzer. In blood vessels of living mouse organs prepared with the IVCT, their RR spectral peaks were also detected at the same RR shifts obtained in human blood. In the blood vessels of living mouse small intestines, some arterioles and venules were clearly distinguishable by monitoring different peak patterns of their RR shifts. The different ratios of the RR shift-areas were calculated even in the arterial vessels. In blood vessels of mouse livers, the Raman spectra showed a lower peak shift of 1378 cm(-1) compared to that of 1355 cm(-1), indicating an SO2 decrease in hepatic blood circulation. Thus, the new cryopreparation technique will enable us to directly analyze the in vivo SO2 in various tissues of a whole animal body prepared with the IVCT, reflecting their living states.


Three-dimensional quantitative structure-activity relationship analysis of inhibitors of human and rat cytochrome P4503A enzymes.
Cytochrome P450 3A4 (CYP3A4) is a member of the CYP family and is an important enzyme in drug metabolism. A compound that inhibits CYP3A4 activity could also affect the pharmacokinetics of other substrates, resulting in drug-drug interactions (DDIs) that could cause side effects. Pharmacokinetic data from drug-development studies in rats often determine the dosage used in human clinical trials. It is therefore useful to understand differences in metabolism in different species at an early stage in drug development. Human and rat CYP3A enzymes show different inhibition profiles with different drugs, although the mechanisms involved are not yet clear. Here we built three-dimensional quantitative structure-activity relationship (3D-QSAR) models using structure-based comparative molecular field analysis (CoMFA), to predict the direct inhibitory activity of ligands for human CYP3A4 and rat CYP3A1, based on computer-ligand docking. The alignment of the ligand docking poses suggested that key amino acid-ligand interactions (e.g., Thr309 in CYP3A4 and Pro310 in CYP3A1) characterized the different potencies with which the ligands inhibited CYP3A4 and CYP3A1. The 3D-QSAR models for human and rat CYP3A family inhibitors predicted the potency of inhibitors and could be useful for assessing DDIs at an early stage in drug discovery.


Effect of focal cerebellar lesions on procedural learning in the serial reaction time task.
Prior studies have shown that procedural learning is severely impaired in patients with diffuse cerebellar damage (cortical degeneration) as measured by the serial reaction time task (SRTT). We hypothesize that focal cerebellar lesions can also have lateralized effects on procedural learning. Our objective was to assess the effects of focal cerebellar lesions in procedural learning as measured by the SRTT. We studied 14 patients with single, unilateral vascular lesions in the territory of the posterior-inferior or superior cerebellar artery, who were compared with ten age- and sex-matched controls in a one-handed version of the SRTT. Patients with lesions at any other level of the brain or posterior fossa were excluded by cranial magnetic resonance imaging. Our results show that patients do not acquire procedural knowledge when performing the task with the hand ipsilateral to the lesion, but show normal learning with the contralateral hand. No correlation was found with the side, size, or vascular territory of the lesion. We conclude that procedural learning is impaired in hemispheric cerebellar lesions and involves only the hand ipsilateral to the lesion, which suggests a critical role for the cerebellum and/or crossed cerebellar-prefrontal connections in this type of learning.


Neuroprotective effects of inositol 1,4,5-trisphosphate receptor C-terminal fragment in a Huntington's disease mouse model.
Huntington's disease (HD) is a dominantly inherited, progressive neurodegenerative disease caused by an expanded polyglutamine tract in huntingtin protein (Htt). Medium spiny striatal neurons (MSNs) are primarily affected in HD. Mutant huntingtin protein (Htt(exp)) specifically binds to and activates type 1 inositol 1,4,5-trisphosphate receptor (InsP(3)R1), an intracellular Ca(2+) release channel. Htt(exp)-InsP(3)R1 association is mediated by a cytosolic C-terminal tail of InsP(3)R1 (a 122-aa-long IC10 fragment). To evaluate an importance of Htt(exp) association with InsP(3)R1 for HD pathology, we generated lentiviral and adeno-associated viruses expressing GFP-IC10 fusion protein and performed a series of experiments with YAC128 HD transgenic mouse. Infection with Lenti-GFP-IC10 virus stabilized Ca(2+) signaling in cultured YAC128 MSNs and protected YAC128 MSNs from glutamate-induced apoptosis. Intrastriatal injections of AAV1-GFP-IC10 significantly alleviated motor deficits and reduced MSN loss and shrinkage in YAC128 mice. Our results demonstrate an importance of InsP(3)R1-Htt(exp) association for HD pathogenesis and suggested that InsP(3)R1 is a potential therapeutic target for HD. Our data also support potential use of IC10 peptide as a novel HD therapeutic agent.


An intelligent rapid odour recognition model in discrimination of Helicobacter pylori and other gastroesophageal isolates in vitro.
Two series of experiments are reported which result in the discrimination between Helicobacter pylori and other bacterial gastroesophageal isolates using a newly developed odour generating system, an electronic nose and a hybrid intelligent odour recognition system. In the first series of experiments, after 5 h of growth (37 degrees C), 53 volatile 'sniffs' were collected over the headspace of complex broth cultures of the following clinical isolates: Staphylococcus aureus, Klebsiella sp., H. pylori, Enterococcus faecalis (10(7) ml(-1)), Mixed infection (Proteus mirabilis, Escherichia coli, and E. faecalis 3 x 10(6) ml each) and sterile cultures. Fifty-six normalised variables were extracted from 14 conductive polymer sensor responses and analysed by a 3-layer back propagation neural network (NN). The NN prediction rate achieved was 98% and the test data (37.7% of all data) was recognised correctly. Successful clustering of bacterial classes was also achieved by discriminant analysis (DA) of a normalised subset of sensor data. Cross-validation identified correctly seven 'unknown' samples. In the second series of experiments after 150 min of microaerobic growth at 37 degrees C, 24 volatile samples were collected over the headspace of H. pylori cultures in enriched (HPP) and normal (HP) media and 11 samples over sterile (N) cultures. Forty-eight sensor parameters were extracted from 12 sensor responses and analysed by a 3-layer NN previously optimised by a genetic algorithm (GA). GA-NN analysis achieved a 94% prediction rate of 'unknown' data. Additionally the 'genetically' selected 16 input neurones were used to perform DA-cross validation that showed a clear clustering of three groups and reclassified correctly nine 'sniffs'. It is concluded that the most important factors that govern the performance of an intelligent bacterial odour detection system are: (a) an odour generation mechanism, (b) a rapid odour delivery system similar to the mammalian olfactory system, (c) a gas sensor array of high reproducibility and (d) a hybrid intelligent model (expert system) which will enable the parallel use of GA-NNs and multivariate techniques.


Inhibition of methane production in microbial fuel cells: operating strategies which select electrogens over methanogens.
Methanogenesis may diminish coulombic efficiency of microbial fuel cells (MFCs), although its importance is application dependent; e.g., suppression of methanogenesis may improve MFC sensing accuracy, but may be tolerable in COD removal from wastewaters. Suppression of methanogenesis was investigated in three H-type MFCs, enriched and acclimated with acetate, propionate and butyrate substrates and subsequently operated under open and closed circuit (OC/CC) regimes. Altering the polarisation state of the electrode displaces microorganisms from the anodic biofilm and leads to observable methane inhibition. The planktonic archeal community was compared to the electrode biofilm whilst under the OC/CC regimes. Semi-quantitative DNA analyses indicate a shift in some dominant species, from the electrode to the solution, during OC operation. The effect of prolonged starvation on anodic species was also studied. The results indicate progressive inhibition of methanogenesis from OC/CC operations; and virtual cessation of methanogenesis when an MFC was starved for a significant period.


Two cases of nevoid basal cell carcinoma syndrome associated with meningioma caused by a PTCH1 or SUFU germline mutation.
Nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder characterized by developmental defects and tumorigenesis. The gene responsible for NBCCS is PTCH1. The development of meningioma in NBCCS patients is a rare event. Here, we report two cases of NBCCS in which meningiomas did develop. The first patient carried a germline mutation in one allele of PTCH1, c.290dupA (p.N97KfsX43). In addition, the meningioma sample carried a somatic mutation, c.307delG (p.Val103LeufsX15), in the other allele of the same gene, suggesting a second hit. This is the first case of NBCCS-associated meningioma explained by the standard two-hit hypothesis. The second patient had a germline nonsense mutation in the SUFU gene, c.550C>T (p.Q184X). SUFU is located downstream of PTCH1 in the sonic hedgehog signaling pathway. This is the second time a germline mutation in SUFU has been found to cause NBCCS. Together with the previous report describing three cases of non-NBCCS medulloblastoma carrying a germline mutation in this gene, individuals with a SUFU germline mutation are expected to have a markedly high risk of developing medulloblastoma and probably meningioma.


Neonatal outcomes following in utero exposure to methadone or buprenorphine: a National Cohort Study of opioid-agonist treatment of Pregnant Women in Norway from 1996 to 2009.
In Norway, most opioid-dependent women are in opioid maintenance treatment (OMT) with either methadone or buprenorphine throughout pregnancy. The inclusion criteria for both medications are the same and both medications are provided by the same health professionals in any part of the country. International studies comparing methadone and buprenorphine in pregnancy have shown differing neonatal outcomes for the two medications. This study compared the neonatal outcomes following prenatal exposure to either methadone or buprenorphine in a national clinical cohort of 139 women/neonates from 1996 to 2009. After adjusting for relevant covariates, buprenorphine-exposed newborns had larger head circumferences and tended to be heavier and longer than methadone-exposed newborns. The incidence of neonatal abstinence syndrome (NAS) and length of treatment of NAS did not differ between methadone- and buprenorphine-exposed newborns. There was little use of illegal drugs and benzodiazepines during the pregnancies. However, the use of any drugs or benzodiazepines during pregnancy was associated with longer lasting NAS-treatment of the neonates. The clinical relevance of these findings is that both methadone and buprenorphine are acceptable medications for the use in pregnancy, in line with previous studies. If starting OMT in pregnancy, buprenorphine should be considered as the drug of choice, due to more favorable neonatal growth parameters. Early confirmation of the pregnancy and systematic follow-up throughout the pregnancy are of importance to encourage the women in OMT to abstain from the use of tobacco, alcohol, illegal drugs or misuse of prescribed drugs.


Identification of a domain within the human T-cell leukemia virus type 2 envelope required for syncytium induction and replication.
In vitro infection by human T-cell leukemia virus type 1 and 2 (HTLV-1 and HTLV-2) can result in syncytium formation, facilitating viral entry. Using cell lines that were susceptible to HTLV-2-mediated syncytium formation but were nonfusogenic with HTLV-1, we constructed chimeric envelopes between HTLV-1 and -2 and assayed for the ability to induce syncytia in BJAB cells and HeLa cells. We have identified a fusion domain composed of the first 64 amino acids at the amino terminus of the HTLV-2 transmembrane protein, p21, the retention of which was required for syncytium induction. Construction of replication-competent HTLV genomic clones allowed us to correlate the ability of HTLV-2 to induce syncytia with the ability to replicate in BJAB cells. Differences in the ability to induce syncytia were not due to differences in the levels of total or cell membrane-associated envelope or in the formation of multimers. Therefore, we have localized a fusion domain within the amino terminus of the transmembrane protein of HTLV-2 envelope that is necessary for syncytium induction and viral replication.


Adsorption and decolorization study of reactive black 5 by immobilized metal-organic framework of UiO-66 and Gloeophyllum trabeum fungus.
This study aimed to investigate immobilized metal-organic framework (MOF) UiO-66 and brown-rot fungus Gloeophyllum trabeum (GT) in PVA-SA matrices for adsorption and decolorization of reactive black 5 (RB5). Furthermore, UiO-66/GT@PVA-SA composite was successfully fabricated and obtained by immobilizing UiO-66 and GT mycelia into a mixture of PVA-SA. This composite demonstrated a decolorization ability of 80.12% for RB5 after 7 days. The composite's reusability was assessed for three cycles; at last, it only achieved 21%. This study reported that adsorption of RB5 by the composite followed a pseudo-second-order kinetic model with a correlation coefficient (R2) of 0.9997. The Freundlich model was found to be suitable for the isotherm adsorption. The process was also spontaneous and feasible, as indicated by the negative ΔG value. Subsequently, four metabolite products resulting from decolorization of RB5 by UiO-66/GT@PVA-SA composite were proposed, namely: C24H19N5Na2O13S4 (m/z = 762), C10H13N2O8S2- (m/z = 353), C12H9N4O7S2- (m/z = 384), and C10H13O8S2- (m/z = 325).


Investigating the interconnectedness of carbon, fossil energy, and financial markets: A dynamic spillover index approach.
Against the background of the global active pursuit of carbon neutrality, this paper uses the DY spillover index method to analyze the spillover network effects between carbon, fossil energy and financial markets. The research results show that the spillover effects between these three markets change over time, with an average spillover index of 25.30%, showing a significant mutual influence. Further analysis found that the EU carbon market plays an important role in spillover effects. Especially under the influence of extreme events, the spillover effects reach their peak. At this time, the degree of mutual influence between markets is as high as 60.01%. In addition, during the COVID-19 epidemic, the spillover effect of the EU carbon market on other markets also reached its maximum, indicating that the epidemic increased the contagion of cross-market risks and caused the carbon market to bear greater risks. The research results of this article have important guiding significance for environmental protection investment and emphasize the importance of formulating differentiated environmental protection policies in different time frames. Facing the dual challenges of global climate change and promoting the goal of carbon neutrality, governments and relevant institutions should pay close attention to changes in spillover effects between markets and timely adjust environmental protection policies to achieve maximum results.


Architecturally diverse proteins converge on an analogous mechanism to inactivate Uracil-DNA glycosylase.
Uracil-DNA glycosylase (UDG) compromises the replication strategies of diverse viruses from unrelated lineages. Virally encoded proteins therefore exist to limit, inhibit or target UDG activity for proteolysis. Viral proteins targeting UDG, such as the bacteriophage proteins ugi, and p56, and the HIV-1 protein Vpr, share no sequence similarity, and are not structurally homologous. Such diversity has hindered identification of known or expected UDG-inhibitory activities in other genomes. The structural basis for UDG inhibition by ugi is well characterized; yet, paradoxically, the structure of the unbound p56 protein is enigmatically unrevealing of its mechanism. To resolve this conundrum, we determined the structure of a p56 dimer bound to UDG. A helix from one of the subunits of p56 occupies the UDG DNA-binding cleft, whereas the dimer interface forms a hydrophobic box to trap a mechanistically important UDG residue. Surprisingly, these p56 inhibitory elements are unexpectedly analogous to features used by ugi despite profound architectural disparity. Contacts from B-DNA to UDG are mimicked by residues of the p56 helix, echoing the role of ugi's inhibitory beta strand. Using mutagenesis, we propose that DNA mimicry by p56 is a targeting and specificity mechanism supporting tight inhibition via hydrophobic sequestration.