Autologous Adrenal Transplantation for the Treatment of Refractory Cushing's Disease.
To report our management of bilateral adrenalectomy with autologous adrenal gland transplantation for persistent Cushing's disease, and to discuss the feasibility of autologous adrenal transplantation for the treatment of refractory Cushing's disease. A retrospective analysis was performed in 4 patients (3 females, aged 14-36 years) who underwent autologous adrenal transplantation for persistent Cushing's disease after endonasal transsphenoidal resection of a pituitary tumor. The procedure was performed by implanting a vascularized adrenal graft into the left iliac fossa with direct and indirect anastomoses. Postoperative follow-up was performed in 1, 1.5, 8, and 10 years, and an over 8-year long-term follow-up was reached in 2 out of the 4 cases. Hormone replacement dosage was guided by clinical symptoms and endocrine results including serum cortisol (F), 24 h urine-free cortisol, and adrenocorticotrophic hormone levels. All 4 patients with symptomatic Cushing's disease experienced resolution of symptoms after autotransplantation without Nelson Syndrome. Functional autografts were confirmed through clinical evaluation and endocrine results. One year after transplantation, adrenal function and hormone replacement dosage remained stable without adrenal hyperplasia. After long-term follow-up, dosages of hormone replacement were reduced in all patients. In this series of 4 patients, we demonstrate the long-term efficacy of bilateral adrenalectomy with autologous adrenal transplantation and propose this procedure as a viable treatment option for refractory Cushing's disease.


Comprehensive Evaluation of Programmed Death-Ligand 1 Expression in Primary and Metastatic Prostate Cancer.
Antibodies targeting the programmed cell death protein 1/programmed death-ligand 1 (PD-L1) interaction have shown clinical activity in multiple cancer types. PD-L1 protein expression is a clinically validated predictive biomarker of response for such therapies. Prior studies evaluating the expression of PD-L1 in primary prostate cancers have reported highly variable rates of PD-L1 positivity. In addition, limited data exist on PD-L1 expression in metastatic castrate-resistant prostate cancer (mCRPC). Here, we determined PD-L1 protein expression by immunohistochemistry using a validated PD-L1-specific antibody (SP263) in a large and representative cohort of primary prostate cancers and prostate cancer metastases. The study included 539 primary prostate cancers comprising 508 acinar adenocarcinomas, 24 prostatic duct adenocarcinomas, 7 small-cell carcinomas, and a total of 57 cases of mCRPC. PD-L1 positivity was low in primary acinar adenocarcinoma, with only 7.7% of cases showing detectable PD-L1 staining. Increased levels of PD-L1 expression were noted in 42.9% of small-cell carcinomas. In mCRPC, 31.6% of cases showed PD-L1-specific immunoreactivity. In conclusion, in this comprehensive evaluation of PD-L1 expression in prostate cancer, PD-L1 expression is rare in primary prostate cancers, but increased rates of PD-L1 positivity were observed in mCRPC. These results will be important for the future clinical development of programmed cell death protein 1/PD-L1-targeting therapies in prostate cancer.


Trial Protocol: Using genotype to tailor prescribing of nicotine replacement therapy: a randomised controlled trial assessing impact of communication upon adherence.
The behavioural impact of pharmacogenomics is untested; informing smokers of genetic test results for responsiveness to smoking cessation medication may increase adherence to this medication. The objective of this trial is to estimate the impact upon adherence to nicotine replacement therapy (NRT) of informing smokers that their oral dose of NRT has been tailored to a DNA analysis. Hypotheses to be tested are as follows: I Adherence to NRT is greater among smokers informed that their oral dose of NRT is tailored to an analysis of DNA (genotype), compared to one tailored to nicotine dependence questionnaire score (phenotype). II Amongst smokers who fail to quit at six months, motivation to make another quit attempt is lower when informed that their oral dose of NRT was tailored to genotype rather than phenotype. An open label, parallel groups randomised trial in which 630 adult smokers (smoking 10 or more cigarettes daily) using National Health Service (NHS) stop smoking services in primary care are randomly allocated to one of two groups:i. NRT oral dose tailored by DNA analysis (OPRM1 gene) (genotype), orii. NRT oral dose tailored by nicotine dependence questionnaire score (phenotype)The primary outcome is proportion of prescribed NRT consumed in the first 28 days following an initial quit attempt, with the secondary outcome being motivation to make another quit attempt, amongst smokers not abstinent at six months. Other outcomes include adherence to NRT in the first seven days and biochemically validated smoking abstinence at six months. The primary outcome will be collected on 630 smokers allowing sufficient power to detect a 7.5% difference in mean proportion of NRT consumed using a two-tailed test at the 5% level of significance between groups. The proportion of all NRT consumed in the first four weeks of quitting will be compared between arms using an independent samples t-test and by estimating the 95% confidence interval for observed between-arm difference in mean NRT consumption (Hypothesis I). Motivation to make another quit attempt will be compared between arms in those failing to quit by six months (Hypothesis II). This is the first clinical trial evaluating the behavioural impact on adherence of prescribing medication using genetic rather than phenotypic information. Specific issues regarding the choice of design for trials of interventions of this kind are discussed. TRIAL DETAILS: Funder: Medical Research Council (MRC)Grant number: G0500274. ISRCTN: 14352545. Date trial stated: June 2007. Expected end date: December 2009. Expected reporting date: December 2010.


Children with dyslexia lack multiple specializations along the visual word-form (VWF) system.
Developmental dyslexia has been associated with a dysfunction of a brain region in the left inferior occipitotemporal cortex, called the "visual word-form area" (VWFA). In adult normal readers, the VWFA is specialized for print processing and sensitive to the orthographic familiarity of letter strings. However, it is still unclear whether these two levels of occipitotemporal specialization are affected in developmental dyslexia. Specifically, we investigated whether (a) these two levels of specialization are impaired in dyslexic children with only a few years of reading experience and (b) whether this impairment is confined to the left inferior occipitotemporal VWFA, or extends to adjacent regions of the "VWF-system" with its posterior-anterior gradient of print specialization. Using fMRI, we measured brain activity in 18 dyslexic and 24 age-matched control children (age 9.7-12.5 years) while they indicated if visual stimuli (real words, pseudohomophones, pseudowords and false-fonts) sounded like a real word. Five adjacent regions of interest (ROIs) in the bilateral occipitotemporal cortex covered the full anterior-posterior extent of the VWF-system. We found that control and dyslexic children activated the same main areas within the reading network. However, a gradient of print specificity (higher anterior activity to letter strings but higher posterior activity to false-fonts) as well as a constant sensitivity to orthographic familiarity (higher activity for unfamiliar than familiar word-forms) along the VWF-system could only be detected in controls. In conclusion, analyzing responses and specialization profiles along the left VWF-system reveals that children with dyslexia show impaired specialization for both print and orthography.


Surgical excision for challenging upper limb nerve sheath tumours: a single centre retrospective review of treatment results.
To review the accuracy of different investigation modalities for upper limb nerve sheath tumours and the resulting surgical outcomes, and propose a standard algorithm to deal with such tumours to minimise complications. Retrospective review. Regional hospital, Hong Kong. All patients with upper limb nerve sheath tumours being excised in our hospital from 1999 to 2008. The accuracy rate of different investigations, as well as corresponding neurological deficits after excision and recurrence rates. A total of 23 (10 male and 13 female) patients, aged between 28 and 72 (mean, 46) years, underwent excision of 25 lesions during the study period. The mean duration of symptom was 2.5 years and tumour size ranged from 1 to 10.5 cm (mean, 2.6 cm). A majority (80%) presented with a typical triad; only one had a true neurological deficit. Twenty-two ultrasonography and 20 magnetic resonance images were obtained, with a diagnostic accuracy of 77% and 100%, respectively. Eight fine-needle aspiration cytology examinations and two core biopsies were performed, which had respective accuracy rates of 13% and 100%. Fifteen patients experienced neurological deficits after the operation; three showed spontaneous recovery. Among 12 patients with long-term residual neurological sequelae, five had both motor and sensory deficits and four had moderate-to-severe disability. No recurrence was reported. Nerve sheath tumours in the hand need to be managed with care. Among the different investigation modalities, magnetic resonance imaging was considered to be the gold standard. Yet ultrasonography is still the most easily accessible and least invasive investigation in public hospital setting. Complications are liable to ensue even if patients are managed by hand specialists. Thus, well-planned operations and detailed discussions with the patient are important prerequisites before operation.


Observational determination of dose-response curves in hallucinogen-treated monkeys.
An objective behavioral profile that was previously shown to distinguish the effects of hallucinogens from those of other classes of drugs was used here to further study hallucinogenic behaviors. Saline, d-amphetamine sulfate and five doses of dimethyltryptamine (DMT) were administered to solitary adolescent rhesus monkeys in a totally dark environment and their behavior was observed via infrared monitors, videotaped, and scored in a number of categories. Scores in the following categories systematically increased with ascending doses of DMT; exploration, locomotion, stereotypy, spasm, tracking and duration of inappropriate behavior. In addition, some behaviors sensitive to hallucinogens occurred with greater frequency in the dark than in a previous study conducted in the light. Behaviors such as tracking and fear grimaces, usually associated with specific stimuli, emerged in the absence of such stimuli in the dark. These results suggested hallucinogen-induced changes in perceptual-motor systems, if not hallucinations per se.


Numerical simulation of two-phase partition chromatography in microchannels for moderated log P measurements.
A finite element simulation has been used in order to study the partition chromatography process of one species between an aqueous mobile phase and an organic stationary phase located at the bottom of a rectangular microchannel. The transient model incorporates convection--diffusion of the species in the water phase coupled to the diffusion in the stationary organic phase by the way of the partition kinetics at the interface. The time evolution of the injected species concentration is analyzed versus the velocity of the mobile phase, the detecting position and the thickness of the stationary phase. The comparison of simulation results with both experimental data and analytical model confirm its validity. These simulations show that thin channels can be used to measure log P of molecules from their retention time. Finally, we have shown how the sample velocity can be optimized for a given geometry of the channel and diffusion coefficient of the species.


Protective Effect of Caffeic Acid against Alzheimer's Disease Pathogenesis via Modulating Cerebral Insulin Signaling, β-Amyloid Accumulation, and Synaptic Plasticity in Hyperinsulinemic Rats.
This study investigated the alleviative effect of caffeic acid (CA) on Alzheimer's disease (AD) pathogenesis and associated mechanisms in high-fat (HF) diet-induced hyperinsulinemic rats. The results of a Morris water maze indicated that, by administrating CA (30 mg/kg b.w./day) for 30 weeks, the memory and learning impairments in HF-induced hyperinsulinemic rats were significantly ameliorated. CA also enhanced superoxide dismutase and glutathione free radical scavenger activity in hyperinsulinemic rats. The Western blot data further confirmed that protein expressions of phosphorylated-glycogen synthase kinase 3β (GSK3β) were significantly increased, whereas the expression of phosphorylated-tau protein decreased in the hippocampus of rats administered with CA in comparison with the HF group. Moreover, the expression of amyloid precursor protein (APP) and β-site APP cleaving enzyme were attenuated, subsequently lowering the level of β-amyloid 1-42 (Aβ 1-42) in the hippocampus of CA-treated hyperinsulinemic rats. CA also significantly increased the expression of synaptic proteins in HF rats.


Genome sequence of the model mushroom Schizophyllum commune.
Much remains to be learned about the biology of mushroom-forming fungi, which are an important source of food, secondary metabolites and industrial enzymes. The wood-degrading fungus Schizophyllum commune is both a genetically tractable model for studying mushroom development and a likely source of enzymes capable of efficient degradation of lignocellulosic biomass. Comparative analyses of its 38.5-megabase genome, which encodes 13,210 predicted genes, reveal the species's unique wood-degrading machinery. One-third of the 471 genes predicted to encode transcription factors are differentially expressed during sexual development of S. commune. Whereas inactivation of one of these, fst4, prevented mushroom formation, inactivation of another, fst3, resulted in more, albeit smaller, mushrooms than in the wild-type fungus. Antisense transcripts may also have a role in the formation of fruiting bodies. Better insight into the mechanisms underlying mushroom formation should affect commercial production of mushrooms and their industrial use for producing enzymes and pharmaceuticals.


Containing the accidental laboratory escape of potential pandemic influenza viruses.
The recent work on the modified H5N1 has stirred an intense debate on the risk associated with the accidental release from biosafety laboratory of potential pandemic pathogens. Here, we assess the risk that the accidental escape of a novel transmissible influenza strain would not be contained in the local community. We develop here a detailed agent-based model that specifically considers laboratory workers and their contacts in microsimulations of the epidemic onset. We consider the following non-pharmaceutical interventions: isolation of the laboratory, laboratory workers' household quarantine, contact tracing of cases and subsequent household quarantine of identified secondary cases, and school and workplace closure both preventive and reactive. Model simulations suggest that there is a non-negligible probability (5% to 15%), strongly dependent on reproduction number and probability of developing clinical symptoms, that the escape event is not detected at all. We find that the containment depends on the timely implementation of non-pharmaceutical interventions and contact tracing and it may be effective (>90% probability per event) only for pathogens with moderate transmissibility (reproductive number no larger than R₀ = 1.5). Containment depends on population density and structure as well, with a probability of giving rise to a global event that is three to five times lower in rural areas. Results suggest that controllability of escape events is not guaranteed and, given the rapid increase of biosafety laboratories worldwide, this poses a serious threat to human health. Our findings may be relevant to policy makers when designing adequate preparedness plans and may have important implications for determining the location of new biosafety laboratories worldwide.


The role of surface ligands in determining the electronic properties of quantum dot solids and their impact on photovoltaic figure of merits.
Surface chemistry plays a crucial role in determining the electronic properties of quantum dot solids and may well be the key to mitigate loss processes involved in quantum dot solar cells. Surface ligands help to maintain the shape and size of the individual dots in solid films, to preserve the clean energy band gap of the individual particles and to control charge carrier conduction across solid films, in turn regulating their performance in photovoltaic applications. In this report, we show that the changes in size, shape and functional groups of small chain organic ligands enable us to modulate mobility, dielectric constant and carrier doping density of lead sulfide quantum dot solids. Furthermore, we correlate these results with performance, stability and recombination processes in the respective photovoltaic devices. Our results highlight the critical role of surface chemistry in the electronic properties of quantum dots. The role of the size, functionality and the surface coverage of the ligands in determining charge transport properties and the stability of quantum dot solids have been discussed. Our findings, when applied in designing new ligands with higher mobility and improved passivation of quantum dot solids, can have important implications for the development of high-performance quantum dot solar cells.


Electrophysiological evidence for phonological priming in Spanish Sign Language lexical access.
Interactive activation models of lexical access assume that the presentation of a given word activates not only its lexical representation but also those corresponding to words similar in form. Current theories are based on data from oral and written languages, and therefore signed languages represent a special challenge for existing theories of word recognition and lexical access since they allow us to question what the genuine fundamentals of human language are and what might be modality-specific adaptation. The aim of the present study is to determine the electrophysiological correlates and time course of phonological processing of Spanish Sign Language (LSE). Ten deaf native LSE signers and ten deaf non-native but highly proficient LSE signers participated in the experiment. We used the ERP methodology and form-based priming in the context of a delayed lexical decision task, manipulating phonological overlap (i.e. related prime-target pairs shared either handshape or location parameters). Results showed that both parameters under study modulated brain responses to the stimuli in different time windows. Phonological priming of location resulted in a higher amplitude of the N400 component (300-500 ms window) for signs but not for non-signs. This effect may be explained in terms of initial competition among candidates. Moreover, the fact that a higher amplitude N400 for related pairs was found for signs but not for non-signs points to an effect at the lexical level. Handshape overlap produced a later effect (600-800 ms window). In this window, a more negative-going wave for the related condition than for the unrelated condition was found for non-signs in the native signers group. The findings are discussed in relation to current models of lexical access and word recognition. Finally, differences between native and non-native signers point to a less efficient use of phonological information among the non-native signers.


Longer pregnancy and slower fetal development in women with latent "asymptomatic" toxoplasmosis.
The purpose of this study was to confirm that women with latent toxoplasmosis have developmentally younger fetuses at estimated pregnancy week 16 and to test four exclusive hypotheses that could explain the observed data. In the present retrospective cohort study we analysed by the GLM (general linear model) method data from 730 Toxoplasma-free and 185 Toxoplasma-infected pregnant women. At pregnancy week 16 estimated from the date of the last menstruation, the mothers with latent toxoplasmosis had developmentally younger fetuses based on ultrasound scan (P = 0.014). Pregnancy of Toxoplasma-positive compared to Toxoplasma-negative women was by about 1.3 days longer, as estimated both from the date of the last menstruation (P = 0.015) and by ultrasonography (P = 0.025). The most parsimonious explanation for the observed data is retarded fetal growth during the first weeks of pregnancy in Toxoplasma-positive women. The phenomenon was only detectable in multiparous women, suggesting that the immune system may play some role in it.


PTSD and Treatment Adherence: the role of health beliefs.
Health beliefs have been shown to influence a myriad of medical treatment decisions. More recently, the impact of health beliefs on treatment decisions for mental illness has become a focus of study. This study examines the health beliefs and treatment behavior of veterans with posttraumatic stress disorder (PTSD). Using standard survey methodology, we assessed beliefs about the cause of PTSD, expected duration and controllability of symptoms, and life consequences of having PTSD. Treatment participation and medication compliance were assessed, as were common treatment correlates, such as patient-provider relationships, dosing frequency, side effect severity, number of prescribed medications, and use of drugs or alcohol to control PTSD symptoms. Explanatory models of PTSD, perceived controllability, and use of benzodiazepines were found to predict psychiatric medication use. Negative life consequences of PTSD were associated with participation in psychotherapy. Assessment of health beliefs may help providers to understand their patients' treatment behavior and to facilitate treatment engagement.


Examining the health effects and bioactive components in Agaricus bisporus mushrooms: a scoping review.
There is evidence from both in vitro and animal models that the consumption of edible mushrooms has beneficial effects on health. It is unclear whether similar effects exist in humans and which bioactive compounds are present. This review synthesises the evidence on the world's most commonly consumed mushroom, Agaricus bisporus to (i) examine its effect on human health outcomes; and (ii) determine the nutrient density of its bioactive compounds, which may explain their health effects. A systematic literature search was conducted on the consumption of A. bisporus, without date and study design limits. Bioactive compounds included ergosterol, ergothioneine, flavonoids, glucans and chitin. Two authors independently identified studies for inclusion and assessed methodological quality. Beneficial effects of A. bisporus on metabolic syndrome, immune function, gastrointestinal health and cancer, with the strongest evidence for the improvement in Vitamin D status in humans, were found. Ultraviolet B (UVB) exposed mushrooms may increase and maintain serum 25(OH)D levels to a similar degree as vitamin D supplements. A. bisporus contain beta-glucans, ergosterol, ergothioneine, vitamin D and an antioxidant compound usually reported as flavonoids; with varying concentrations depending on the type of mushroom, cooking method and duration, and UVB exposure. Further research is required to fully elucidate the bioactive compounds in mushrooms using vigorous analytical methods and expand the immunological markers being tested. To enable findings to be adopted into clinical practice and public health initiatives, replication of existing studies in different population groups is required to confirm the impact of A. bisporus on human health.


Adult-Onset Autoimmune Enteropathy in an European Tertiary Referral Center.
Adult-onset autoimmune enteropathy (AIE) is a rare cause of severe chronic diarrhea because of small intestinal villous atrophy. We report on patients with adult-onset AIE in an European referral center. Retrospective study including patients diagnosed with AIE in the Amsterdam UMC, location VUmc, between January 2003 and December 2019. Clinical, serological, and histological features and response to treatment were reported. The specificity of antienterocyte antibodies (AEA) was evaluated by examining the prevalence of AEA in (i) controls (n = 30) and in patients with (ii) AIE (n = 13), (iii) celiac disease (CD, n = 52), (iv) refractory celiac disease type 2 (n = 18), and (v) enteropathy-associated T-cell lymphoma (EATL, n = 10). Thirteen AIE patients were included, 8 women (62%), median age of 52 years (range 23-73), and 6 (46%) with an autoimmune disease. AEA were observed in 11 cases (85%), but were also found in CD (7.7%), refractory celiac disease type 2 (16.7%), and EATL (20%). Ten patients (77%) were human leukocyte antigen DQ2.5 heterozygous. Total parenteral nutrition was required in 8 cases (62%). Steroids induced clinical remission in 8 cases (62%). Step-up therapy with rituximab, cyclosporine, infliximab, and cladribine in steroid-refractory patients was only moderately effective. Four patients died (31%), but 4 (31%) others are in long-term drug-free remission after receiving immunosuppressive treatment, including 1 patient who underwent autologous stem cell transplantation. Adult-onset AIE is a rare but severe enteropathy that occurs in patients susceptible for autoimmune disease. Four patients (31%) died secondary to therapy-refractory malabsorption, while immunosuppressive therapy leads to a long-lasting drug-free remission in one-third of patients.


Utility of Human Relevant Preclinical Animal Models in Navigating NAFLD to MAFLD Paradigm.
Fatty liver disease is an emerging contributor to disease burden worldwide. The past decades of work established the heterogeneous nature of non-alcoholic fatty liver disease (NAFLD) etiology and systemic contributions to the pathogenesis of the disease. This called for the proposal of a redefinition in 2020 to that of metabolic dysfunction-associated fatty liver disease (MAFLD) to better reflect the current understanding of the disease. To date, several clinical cohort studies comparing NAFLD and MAFLD hint at the relevancy of the new nomenclature in enriching for patients with more severe hepatic injury and extrahepatic comorbidities. However, the underlying systemic pathogenesis is still not fully understood. Preclinical animal models have been imperative in elucidating key biological mechanisms in various contexts, including intrahepatic disease progression, interorgan crosstalk and systemic dysregulation. Furthermore, they are integral in developing novel therapeutics against MAFLD. However, substantial contextual variabilities exist across different models due to the lack of standardization in several aspects. As such, it is crucial to understand the strengths and weaknesses of existing models to better align them to the human condition. In this review, we consolidate the implications arising from the change in nomenclature and summarize MAFLD pathogenesis. Subsequently, we provide an updated evaluation of existing MAFLD preclinical models in alignment with the new definitions and perspectives to improve their translational relevance.


Comparison between automatic tube compensation and continuous positive airway pressure during spontaneous breathing trials.
Various methods to perform spontaneous breathing trials (SBTs) exist, but no one method has been shown to be superior. Automatic tube compensation (ATC) is a new and potentially advantageous ventilation mode to use during SBT. We compared ATC to continuous positive airway pressure (CPAP) during SBTs, to determine their efficacy in identifying patients ready to be liberated from mechanical ventilation. We randomized 118 adults in a general intensive care unit on mechanical ventilation for > or = 24 h who were about to undergo an SBT as part of an established respiratory-therapist-driven weaning protocol to undergo 30 min SBT with ATC or CPAP with no pressure support. We predefined the SBT-failure criteria. The primary outcome was duration of weaning (days from first SBT to extubation). Other outcomes included unsuccessful extubation within 48 h, first-SBT-pass rate, and total duration of mechanical ventilation. We found a trend toward less failure of first SBT with ATC, compared to CPAP (3% vs 13% respectively, P = .09), but no difference in duration of weaning, rate of unsuccessful extubation, or duration of mechanical ventilation. When applied as part of a respiratory-therapist-driven weaning protocol in a general intensive-care population, SBTs with ATC were safe but did not hasten liberation from mechanical ventilation, when compared to CPAP.


Object representations at multiple scales from digital elevation models.
In the last decade landform classification and mapping has developed as one of the most active areas of geomorphometry. However, translation from continuous models of elevation and its derivatives (slope, aspect, and curvatures) to landform divisions (landforms and landform elements) is filtered by two important concepts: scale and object ontology. Although acknowledged as being important, these two issues have received surprisingly little attention.This contribution provides an overview and prospects of object representation from DEMs as a function of scale. Relationships between object delineation and classification or regionalization are explored, in the context of differences between general and specific geomorphometry. A review of scales issues in geomorphometry-ranging from scale effects to scale optimization techniques-is followed by an analysis of pros and cons of using cells and objects in DEM analysis. Prospects for coupling multi-scale analysis and object delineation are then discussed. Within this context, we propose discrete geomorphometry as a possible approach between general and specific geomorphometry. Discrete geomorphometry would apply to and describe land-surface divisions defined solely by the criteria of homogeneity in respect to a given land-surface parameter or a combination of several parameters. Homogeneity, in its turn, should always be relative to scale.


Cognitive control predicts use of model-based reinforcement learning.
Accounts of decision-making and its neural substrates have long posited the operation of separate, competing valuation systems in the control of choice behavior. Recent theoretical and experimental work suggest that this classic distinction between behaviorally and neurally dissociable systems for habitual and goal-directed (or more generally, automatic and controlled) choice may arise from two computational strategies for reinforcement learning (RL), called model-free and model-based RL, but the cognitive or computational processes by which one system may dominate over the other in the control of behavior is a matter of ongoing investigation. To elucidate this question, we leverage the theoretical framework of cognitive control, demonstrating that individual differences in utilization of goal-related contextual information--in the service of overcoming habitual, stimulus-driven responses--in established cognitive control paradigms predict model-based behavior in a separate, sequential choice task. The behavioral correspondence between cognitive control and model-based RL compellingly suggests that a common set of processes may underpin the two behaviors. In particular, computational mechanisms originally proposed to underlie controlled behavior may be applicable to understanding the interactions between model-based and model-free choice behavior.


Comparison of imputation methods for handling missing covariate data when fitting a Cox proportional hazards model: a resampling study.
The appropriate handling of missing covariate data in prognostic modelling studies is yet to be conclusively determined. A resampling study was performed to investigate the effects of different missing data methods on the performance of a prognostic model. Observed data for 1000 cases were sampled with replacement from a large complete dataset of 7507 patients to obtain 500 replications. Five levels of missingness (ranging from 5% to 75%) were imposed on three covariates using a missing at random (MAR) mechanism. Five missing data methods were applied; a) complete case analysis (CC) b) single imputation using regression switching with predictive mean matching (SI), c) multiple imputation using regression switching imputation, d) multiple imputation using regression switching with predictive mean matching (MICE-PMM) and e) multiple imputation using flexible additive imputation models. A Cox proportional hazards model was fitted to each dataset and estimates for the regression coefficients and model performance measures obtained. CC produced biased regression coefficient estimates and inflated standard errors (SEs) with 25% or more missingness. The underestimated SE after SI resulted in poor coverage with 25% or more missingness. Of the MI approaches investigated, MI using MICE-PMM produced the least biased estimates and better model performance measures. However, this MI approach still produced biased regression coefficient estimates with 75% missingness. Very few differences were seen between the results from all missing data approaches with 5% missingness. However, performing MI using MICE-PMM may be the preferred missing data approach for handling between 10% and 50% MAR missingness.


Blockage of melatonin receptors impairs p53-mediated prevention of DNA damage accumulation.
Melatonin has been known to be a chemopreventive agent since its levels inversely correlate with the risk of developing cancer. We have recently shown that melatonin induces p38-dependent phosphorylation of both p53 and histone H2AX. This is associated with a p53-mediated increase in repair of both endogenous and chemotherapy-induced DNA damage. In addition, the inhibition of p38 activities impairs melatonin's capability to induce a p53-dependent DNA damage response and thus its ability to maintain genome integrity. Since melatonin-induced p53 phosphorylation requires an intact p38 phosphorylation cascade and p38 can be activated by G proteins, we supposed that melatonin's activities could be mediated by its G-protein-coupled membrane receptors, MT1 and MT2. Here, we show that the activation of the p53-dependent DNA damage response by melatonin is indeed mediated by MT1 and MT2. As a result, the absence of either receptor impairs melatonin's ability to reduce both cell proliferation and clonogenic potential of cancer cells. In addition, this causes an impairment of the p53-dependent DNA damage response. By providing molecular insight, our findings might have translational impact, suggesting the involvement of melatonin receptors in tumorigenesis.


Functional proteomic profiling of AML predicts response and survival.
Because protein function regulates the phenotypic characteristics of cancer, a functional proteomic classification system could provide important information for pathogenesis and prognosis. With the goal of ultimately developing a proteomic-based classification of acute myeloid leukemia (AML), we assayed leukemia-enriched cells from 256 newly diagnosed AML patients, for 51 total and phosphoproteins from apoptosis, cell-cycle, and signal-transduction pathways, using reverse-phase protein arrays. Expression in matched blood and marrow samples were similar for 44 proteins; another 7 had small fold changes (8%-55%), suggesting that functional proteomics of leukemia-enriched cells in the marrow and periphery are similar. Protein expression patterns were independent of clinical characteristics. However, 24 proteins were significantly different between French-American-British subtypes, defining distinct signatures for each. Expression signatures for AML with cytogenetic abnormalities involving -5 or -7 were similar suggesting mechanistic commonalities. Distinct expression patterns for FMS-like tyrosine kinase 3-internal tandem duplication were also identified. Principal component analysis defined 7 protein signature groups, with prognostic information distinct from cytogenetics that correlated with remission attainment, relapse, and overall survival. In conclusion, protein expression profiling patterns in AML correlate with known morphologic features, cytogenetics, and outcome. Confirmation in independent studies may also provide pathophysiologic insights facilitating triage of patients to emerging targeted therapies.


Improving physiotherapy services to Indigenous children with physical disability: Are client perspectives missed in the continuous quality improvement approach?
To compare the outcomes of two cycles of continuous quality improvement (CQI) at a paediatric physiotherapy service with findings from interviews with clients and their carers using the service. Case study based at one paediatric physiotherapy service Community-based paediatric allied health service in Northern Australia. Forty-nine clinical records and four staff at physiotherapy service, five Indigenous children with physical disability aged 8-21 years, and nine carers of Indigenous children aged 0-21 years (current or previous clients). The CQI process based on the Audit and Best Practice for Chronic Disease involved a clinical audit; a workshop where clinicians assessed their health care systems, identified weaknesses and strengths, and developed goals and strategies for improvement; and reassessment through a second audit and workshop. Twelve open-ended, in-depth interviews were conducted with previous or current clients selected through purposive and theoretical sampling. CQI and interview results were then compared. Comparison of findings from the two studies Both CQI and interview results highlighted service delivery flexibility and therapists' knowledge, support and advocacy as service strengths, and lack of resources and a child-friendly office environment as weaknesses. However, the CQI results reported better communication and client input into the service than the interview results. The CQI process, while demonstrating improvements in clinical and organisational aspects of the service, did not always reflect or address the primary concerns of Indigenous clients and underlined the importance of including clients in the CQI process.


Bone morphogenetic proteins induce apoptosis in human pulmonary vascular smooth muscle cells.
Pulmonary vascular medial hypertrophy in primary pulmonary hypertension (PPH) is mainly caused by increased proliferation and decreased apoptosis in pulmonary artery smooth muscle cells (PASMCs). Mutations of the bone morphogenetic protein (BMP) receptor type II (BMP-RII) gene have been implicated in patients with familial and sporadic PPH. The objective of this study was to elucidate the apoptotic effects of BMPs on normal human PASMCs and to examine whether BMP-induced effects are altered in PASMCs from PPH patients. Using RT-PCR, we detected six isoforms of BMPs (BMP-1 through -6) and three subunits of BMP receptors (BMP-RIa, -RIb, and -RII) in PASMCs. Treatment of normal PASMCs with BMP-2 or -7 (100-200 nM, 24-48 h) markedly increased the percentage of cells undergoing apoptosis. The BMP-2-mediated apoptosis in normal PASMCs was associated with a transient activation or phosphorylation of Smad1 and a marked downregulation of the antiapoptotic protein Bcl-2. In PASMCs from PPH patients, the BMP-2- or BMP-7-induced apoptosis was significantly inhibited compared with PASMCs from patients with secondary pulmonary hypertension. These results suggest that the antiproliferative effect of BMPs is partially due to induction of PASMC apoptosis, which serves as a critical mechanism to maintain normal cell number in the pulmonary vasculature. Inhibition of BMP-induced PASMC apoptosis in PPH patients may play an important role in the development of pulmonary vascular medial hypertrophy in these patients.


Gaussian graphical modeling reconstructs pathway reactions from high-throughput metabolomics data.
With the advent of high-throughput targeted metabolic profiling techniques, the question of how to interpret and analyze the resulting vast amount of data becomes more and more important. In this work we address the reconstruction of metabolic reactions from cross-sectional metabolomics data, that is without the requirement for time-resolved measurements or specific system perturbations. Previous studies in this area mainly focused on Pearson correlation coefficients, which however are generally incapable of distinguishing between direct and indirect metabolic interactions. In our new approach we propose the application of a Gaussian graphical model (GGM), an undirected probabilistic graphical model estimating the conditional dependence between variables. GGMs are based on partial correlation coefficients, that is pairwise Pearson correlation coefficients conditioned against the correlation with all other metabolites. We first demonstrate the general validity of the method and its advantages over regular correlation networks with computer-simulated reaction systems. Then we estimate a GGM on data from a large human population cohort, covering 1020 fasting blood serum samples with 151 quantified metabolites. The GGM is much sparser than the correlation network, shows a modular structure with respect to metabolite classes, and is stable to the choice of samples in the data set. On the example of human fatty acid metabolism, we demonstrate for the first time that high partial correlation coefficients generally correspond to known metabolic reactions. This feature is evaluated both manually by investigating specific pairs of high-scoring metabolites, and then systematically on a literature-curated model of fatty acid synthesis and degradation. Our method detects many known reactions along with possibly novel pathway interactions, representing candidates for further experimental examination. In summary, we demonstrate strong signatures of intracellular pathways in blood serum data, and provide a valuable tool for the unbiased reconstruction of metabolic reactions from large-scale metabolomics data sets.


The State of Open Source Electronic Health Record Projects: A Software Anthropology Study.
Electronic health records (EHR) are a key tool in managing and storing patients' information. Currently, there are over 50 open source EHR systems available. Functionality and usability are important factors for determining the success of any system. These factors are often a direct reflection of the domain knowledge and developers' motivations. However, few published studies have focused on the characteristics of free and open source software (F/OSS) EHR systems and none to date have discussed the motivation, knowledge background, and demographic characteristics of the developers involved in open source EHR projects. This study analyzed the characteristics of prevailing F/OSS EHR systems and aimed to provide an understanding of the motivation, knowledge background, and characteristics of the developers. This study identified F/OSS EHR projects on SourceForge and other websites from May to July 2014. Projects were classified and characterized by license type, downloads, programming languages, spoken languages, project age, development status, supporting materials, top downloads by country, and whether they were "certified" EHRs. Health care F/OSS developers were also surveyed using an online survey. At the time of the assessment, we uncovered 54 open source EHR projects, but only four of them had been successfully certified under the Office of the National Coordinator for Health Information Technology (ONC Health IT) Certification Program. In the majority of cases, the open source EHR software was downloaded by users in the United States (64.07%, 148,666/232,034), underscoring that there is a significant interest in EHR open source applications in the United States. A survey of EHR open source developers was conducted and a total of 103 developers responded to the online questionnaire. The majority of EHR F/OSS developers (65.3%, 66/101) are participating in F/OSS projects as part of a paid activity and only 25.7% (26/101) of EHR F/OSS developers are, or have been, health care providers in their careers. In addition, 45% (45/99) of developers do not work in the health care field. The research presented in this study highlights some challenges that may be hindering the future of health care F/OSS. A minority of developers have been health care professionals, and only 55% (54/99) work in the health care field. This undoubtedly limits the ability of functional design of F/OSS EHR systems from being a competitive advantage over prevailing commercial EHR systems. Open source software seems to be a significant interest to many; however, given that only four F/OSS EHR systems are ONC-certified, this interest is unlikely to yield significant adoption of these systems in the United States. Although the Health Information Technology for Economic and Clinical Health (HITECH) act was responsible for a substantial infusion of capital into the EHR marketplace, the lack of a corporate entity in most F/OSS EHR projects translates to a marginal capacity to market the respective F/OSS system and to navigate certification. This likely has further disadvantaged F/OSS EHR adoption in the United States.


Yeast enhancer of polycomb defines global Esa1-dependent acetylation of chromatin.
Drosophila Enhancer of Polycomb, E(Pc), is a suppressor of position-effect variegation and an enhancer of both Polycomb and trithorax mutations. A homologous yeast protein, Epl1, is a subunit of the NuA4 histone acetyltransferase complex. Epl1 depletion causes cells to accumulate in G2/M and global loss of acetylated histones H4 and H2A. In relation to the Drosophila protein, mutation of Epl1 suppresses gene silencing by telomere position effect. Epl1 protein is found in the NuA4 complex and a novel highly active smaller complex named Piccolo NuA4 (picNuA4). The picNuA4 complex contains Esa1, Epl1, and Yng2 as subunits and strongly prefers chromatin over free histones as substrate. Epl1 conserved N-terminal domain bridges Esa1 and Yng2 together, stimulating Esa1 catalytic activity and enabling acetylation of chromatin substrates. A recombinant picNuA4 complex shows characteristics similar to the native complex, including strong chromatin preference. Cells expressing only the N-terminal half of Epl1 lack NuA4 HAT activity, but possess picNuA4 complex and activity. These results indicate that the essential aspect of Esa1 and Epl1 resides in picNuA4 function. We propose that picNuA4 represents a nontargeted histone H4/H2A acetyltransferase activity responsible for global acetylation, whereas the NuA4 complex is recruited to specific genomic loci to perturb locally the dynamic acetylation/deacetylation equilibrium.


Neuroprotective effects of resveratrol and epigallocatechin gallate polyphenols are mediated by the activation of protein kinase C gamma.
Polyphenols such as epigallocatechin gallate (EGCG) and resveratrol have received a great deal of attention because they may contribute to the purported neuroprotective action of the regular consumption of green tea and red wine. Many studies, including those published by our group, suggest that this protective action includes their abilities to prevent the neurotoxic effects of beta-amyloid, a protein whose accumulation likely plays a pivotal role in Alzheimer's disease. Moreover, the scavenging activities of polyphenols on reactive oxygen species and their inhibitory action of cyclooxygenase likely explain, at least in part, their antioxidant and anti-inflammatory activities. Besides these well-documented properties, the modulatory action of these polyphenols on intracellular signaling pathways related to cell death/survival (e.g., protein kinase C, PKC) has yet to be investigated in detail. Using rat hippocampal neuronal cells, we aimed to investigate here the effects of EGCG and resveratrol on cell death induced by GF 109203X, a selective inhibitor of PKC. The MTT/resazurin and spectrin assays indicated that EGCG and resveratrol protected against GF 109203X-induced cell death and cytoskeleton degeneration, with a maximal effect at 1 and 3 μM, respectively. Moreover, immunofluorescence data revealed that cells treated with these polyphenols increased PKC gamma (γ) activation and promoted neuronal interconnections. Finally, we found that the protective effects of both polyphenols on the cytoskeleton and synaptic plasticity were mediated by the PKCγ subunit. Taken together, the results suggest that PKC, and more specifically its γ subunit, plays a critical role in the protective action of EGCG and resveratrol on neuronal integrity.


Phytochemical screening and acute toxicity evaluation of Telfairia occidentalis aqueous extracts on rats.
The phytochemical composition and acute toxicity of Telfairia occidentalis aqueous extracts were investigated in this study. Phytochemical screening was carried out on the pulverized leaf, root, pod and stem samples. Proximate analysis was also conducted for the root to ascertain the effect of drying procedures on its composition. Fifty-six (56) Wister albino rats, male and female were divided into two broad groups of 28 animals per group. The first group was randomly separated into seven (7) groups of four (4) animals per group. The control group received distilled water alone while the other groups received varied doses (1500mg/kg, 2250mg/kg and 3000mg/kg) of the Soluble and Insoluble Tefairia occidentalis root fraction. The second group of 28 animals was also distributed into 7 groups of 4 animals per group. Six test groups received varied doses (1500mg/kg, 2250mg/kg and 3000mg/kg) of Telfairia occidentalis fruit and stem extracts. The animals were observed for the first 12hr for any toxic symptoms and for 48 hr for mortality rate. Surviving animals were sacrificed after 48 hours. Phytochemical screening results reveal the presence of tannins, flavonoid, steroid, terpenoids, saponin, alkaloid, glycosides, proteins and carbohydrates. Flavonoid and saponin was not detected in stem sample; alkaloid is present in all samples except pod; and cyanogenic glycoside was found in both root and pod samples. Except for the fibre content, the method of preparation of the root had no significant effect on the proximate composition of the sample. The root extracts cause insignificant reduction in Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) activities, except for the significant reduction in ALT activity at highest dose. The pod extract significantly increased the ALT and AST activities, which is dose dependent, while the stem extract only caused increased activity of ALT, but not AST. None of the extracts administered had any significant effect on the levels of serum creatinine and urea. Thus, while the root extract may exhibit some hepatoprotective effect (or nephrotoxic due to cyanogenic glycoside) and its proximate composition, not affected by heat treatment, the pod and stem extracts of Telfairia occidentalis may have some effects on rat hepatocytes.


Enhancer hubs and loop collisions identified from single-allele topologies.
Chromatin folding contributes to the regulation of genomic processes such as gene activity. Existing conformation capture methods characterize genome topology through analysis of pairwise chromatin contacts in populations of cells but cannot discern whether individual interactions occur simultaneously or competitively. Here we present multi-contact 4C (MC-4C), which applies Nanopore sequencing to study multi-way DNA conformations of individual alleles. MC-4C distinguishes cooperative from random and competing interactions and identifies previously missed structures in subpopulations of cells. We show that individual elements of the β-globin superenhancer can aggregate into an enhancer hub that can simultaneously accommodate two genes. Neighboring chromatin domain loops can form rosette-like structures through collision of their CTCF-bound anchors, as seen most prominently in cells lacking the cohesin-unloading factor WAPL. Here, massive collision of CTCF-anchored chromatin loops is believed to reflect 'cohesin traffic jams'. Single-allele topology studies thus help us understand the mechanisms underlying genome folding and functioning.


Suppression of spermatogenesis in man induced by Nal-Glu gonadotropin releasing hormone antagonist and testosterone enanthate (TE) is maintained by TE alone.
GnRH antagonists plus testosterone (T) suppress LH and FSH levels and inhibit spermatogenesis to azoospermia or severe oligozoospermia. High-dose T treatment alone has been shown to be an effective male contraceptive (contraceptive efficacy rate of 1.4 per 100 person yr). Combined GnRH antagonist and T induces azoospermia more rapidly and at a higher incidence than T alone; this combination has therefore been proposed as a prototype male contraceptive. However, because GnRH antagonists are expensive to synthesize and difficult to deliver, it would be desirable to rapidly suppress sperm counts to low levels with GnRH antagonist plus T and maintain azoospermia or severe oligozoospermia with T alone. In this study, 15 healthy men (age 21-41 yr) with normal semen analyses were treated with T enanthate (TE) 100 mg im/week plus 10 mg Nal-Glu GnRH antagonist sc daily for 12 weeks to induce azoospermia or severe oligozoospermia. At 12-16 weeks, 10 of 15 subjects had zero sperm counts, and 14 of 15 had sperm counts less than 3 x 10(6)/mL. The 14 who were suppressed on combined treatment were maintained on TE alone (100 mg/week im) for an additional 20 weeks. Thirteen of 14 subjects in the TE alone phase had sperm counts maintained at less than 3 x 10(6)/mL for 20 weeks. Ten remained persistently azoospermic or had sperm concentration of 0.1 x 10(6)/mL once during maintenance. Mean LH and FSH levels in the subjects were suppressed to 0.4+/-0.2 IU/L and 0.5+/-0.2 IU/L in the induction phase, which was maintained in the maintenance phase. The 1 subject who failed to suppress sperm counts during induction had serum LH and FSH reduced to 0.3 and 0.5 IU/L, respectively. The subject who failed to maintenance had LH and FSH suppressed to 1.0 and 0.2 IU/L, respectively, during the induction phase but these rose to 1.6 and 2.1 IU/L, respectively, during maintenance. Failure to suppress or maintain low sperm counts may be related to incomplete suppression of serum LH and FSH levels. We conclude that sperm counts suppressed with GnRH antagonist plus T can be maintained with relatively low dose TE treatment alone. This concept should be explored further in the development of effective, safe, and affordable hormonal male contraceptives.


The inventory of parent and peer attachment: Individual differences and their relationship to psychological well-being in adolescence.
The results of two studies are reported. Study I involved the development of the Inventory of Parent and Peer Attachment (IPPA), a self-report instrument for use with adolescents. Subject were 179 college students aged 16-20 years. Item content of the instrument was suggested by attachment theory's formulations concerning the nature of feelings toward attachment figures. In Study II, the convergent validity of the IPPA was examined. Also, a hierarchial regression model was employed to investigate the association between quality of attachment and self-esteem, life-satisfaction, and affective status. Respondents were 86 adolescents from the Study I sample. As hypothesized, perceived quality of both parent and peer attachments was significantly related to psychological well-being. Results of the development of a theoretically focused, exploratory classification scheme indicated that adolescents classified as highly securely attached reported greater satisfaction with themselves, a higher likelihood of seeking social support, and less symptomatic response to stressful life events.


Assembling and ordering polymer-grafted nanoparticles in three dimensions.
Taking advantage of the aqueous biphasic behavior of polyethylene glycol (PEG)/salts, recent experiments have demonstrated self-assembly and crystallization of PEG-grafted gold nanoparticles (PEG-AuNPs) into tunable two-dimensional (2D) supercrystals by adjusting salt concentration (for instance, K2CO3). In those studies, combined experimental evidence and theoretical analysis have pointed out the possibility that similar strategies can lead to three-dimensional (3D) formation of ordered nanoparticle precipitates. Indeed, a detailed small-angle X-ray scattering (SAXS) study reported herein reveals the spontaneous formation of PEG-AuNPs assemblies in high-concentration salt solutions that exhibit short-range 3D order compatible with fcc symmetry. We argue that the assembly into fcc crystals is driven by partnering nearest-neighbors to minimize an effective surface-tension gradient at the boundary between the polymer shell and the high-salt media. We report SAXS and other results on PEG-AuNPs of various Au core diameters in the range of 10 to 50 nm and analyze them in the framework of brush-polymer theory revealing a systematic prediction of the nearest-neighbor distance in the 3D assemblies.


A novel abundance-based algorithm for binning metagenomic sequences using l-tuples.
Metagenomics is the study of microbial communities sampled directly from their natural environment, without prior culturing. Among the computational tools recently developed for metagenomic sequence analysis, binning tools attempt to classify the sequences in a metagenomic dataset into different bins (i.e., species), based on various DNA composition patterns (e.g., the tetramer frequencies) of various genomes. Composition-based binning methods, however, cannot be used to classify very short fragments, because of the substantial variation of DNA composition patterns within a single genome. We developed a novel approach (AbundanceBin) for metagenomics binning by utilizing the different abundances of species living in the same environment. AbundanceBin is an application of the Lander-Waterman model to metagenomics, which is based on the l-tuple content of the reads. AbundanceBin achieved accurate, unsupervised, clustering of metagenomic sequences into different bins, such that the reads classified in a bin belong to species of identical or very similar abundances in the sample. In addition, AbundanceBin gave accurate estimations of species abundances, as well as their genome sizes-two important parameters for characterizing a microbial community. We also show that AbundanceBin performed well when the sequence lengths are very short (e.g., 75 bp) or have sequencing errors. By combining AbundanceBin and a composition-based method (MetaCluster), we can achieve even higher binning accuracy. Supplementary Material is available at www.liebertonline.com/cmb .


It's only teeth - are there limits to genetic testing?
Dental genetic disorders can cause severe social and psychological effects in affected individuals. The cost of treatment can be considerable, not only in financial terms but also in time spent during treatment. In theory it is, or will soon be, possible to use advances in molecular genetics for pre-natal testing, for selection of embryos using in vitro fertilization techniques, and for gene therapy. The questions we pose are whether these approaches are appropriate. We hope that this review will stimulate debate on these issues.


Modification and uptake of a cisplatin carbonato complex by Jurkat cells.
The interactions of Jurkat cells with cisplatin, cis-[Pt(15NH3)2Cl2]1, are studied using 1H-15N heteronuclear single quantum coherence (HSQC) NMR and inductively coupled plasma mass spectrometry. We show that Jurkat cells in culture rapidly modify the monocarbonato complex cis-[Pt(15NH3)2(CO3)Cl]- (4), a cisplatin species that forms in culture media and probably also in blood. Analysis of the HSQC NMR peak intensity for 4 in the presence of different numbers of Jurkat cells reveals that each cell is capable of modifying 0.0028 pmol of 4 within approximately 0.6 h. The amounts of platinum taken up by the cell, weakly bound to the cell surface, remaining in the culture medium, and bound to genomic DNA were measured as functions of time of exposure to different concentrations of drug. The results show that most of the 4 that has been modified by the cells remains in the culture medium as a substance of molecular mass <3 kDa, which is HSQC NMR silent, and is not taken up by the cell. These results are consistent with a hitherto undocumented extracellular detoxification mechanism in which the cells rapidly modify 4, which is present in the culture medium, so it cannot bind to the cell. Because there is only a slow decrease in the amount of unmodified 4 remaining in the culture medium after 1 h, -1.1 +/- 0.4 microM h(-1), the cells subsequently lose their ability to modify 4. These observations have important implications for the mechanism of action of cisplatin.


Detection of reverse transcriptase activity in human melanoma cell lines and identification of a murine leukemia virus contaminant.
Stimulated by earlier reports on the presence of retroviruses in mouse and hamster melanoma cell lines, we addressed the question as to whether human melanoma cell lines might also harbour a retrovirus. The melanoma cell lines SK-MEL-25, SK-MEL-28, MEL-JUSO, MML-I, MeWo, A-375, Colo-38, BS-780 were confirmed to be human by human leucocyte antigen (HLA) typing, and supernatants were tested by the product-enhanced reverse transcriptase (PERT) assay for reverse transcriptase (RT) activity. Cell lines SK-MEL-25, SK-MEL-28, MEL-JUSO and MML-I were positive, whereas cell lines MeWo, A-375, Colo-38 and BS-780 were negative. The RT activity peaked at a buoyant density in sucrose typical for retroviruses. From this peak fraction an R-U5 sequence indistinguishable from murine leukemia virus (MLV) was identified by particle-associated retrovirus RNA amplification (PARRA). Semiquantitative MLV-specific RNA-PCR demonstrated colocalization of the MLV-like RNA and RT activity on the sucrose gradient of SK-Mel-25. MLV RNA and DNA were also detectable in culture supernatants of SK-MEL-28, MEL-JUSO and MML-I, but not of MeWo, A-375, Colo-38 and BS-780 by semiquantitative polymerase chain reaction (PCR). Sequence comparison revealed highest homology with the RET sequence previously identified in mouse myeloma SP2/0-AG14 cells. Taken together, our data strongly suggest that certain human melanoma cell lines are productively infected by a MLV which was probably introduced during tumour passage in mice or by laboratory contamination many years ago and subsequently spread to other lines. We recommend mandatory testing of melanoma and other human cell lines for contamination with infectious MLV or other animal retroviruses, similar to mycoplasma screening, in order to avoid artificial experimental data.


Disc distraction shows evidence of regenerative potential in degenerated intervertebral discs as evaluated by protein expression, magnetic resonance imaging, and messenger ribonucleic acid expression analysis.
An animal model of degeneration was used to determine the effects of disc distraction, and was evaluated with magnetic resonance imaging (MRI) as well as gene and protein expression levels. To investigate gene expression and MRI effects of distraction. Disc degeneration can result from hyper-physiologic loading. Distracted discs with degeneration showed histologic signs of tissue recovery. There were 18 rabbits that underwent 28 days of compression (200 N) to induce moderate disc degeneration followed by 28 days of distraction (120 N; attached and loaded distraction device) or sham distraction (attached but unloaded distraction device). Comparison was performed with 56 days of compressed discs without distraction. Quantitative outcome measures were MRI signal intensity and gene expression analysis to determine: messenger ribonucleic acid levels for extracellular matrix genes, including collagen 1, collagen 2, biglycan, decorin, aggrecan, fibromodulin, and osteonectin; and matrix-regulative genes, including matrix metalloproteinase-13, tissue-inhibitor of matrix metalloproteinase-1, and bone morphogenetic protein (BMP)-2. Immunohistology was performed for collagen 2 and BMP-2 to label cells semiquantitatively by staining of the cell-surrounding matrix. A total of 28 days of compression decreased signal intensity. Distraction over the same period reestablished physiologic signal intensity, however, a persistent reduction was found in sham distraction. Distraction resulted in gene expression up-regulation of collagen 1 (5.4-fold), collagen 2 (5.5-fold), biglycan (7.7-fold), and decorin (3.4-fold), while expression of fibromodulin (0.16-fold), tissue-inhibitor of matrix metalloproteinase-1 (0.05-fold), and BMP-2 (0.15-fold) was decreased, as compared with 56 days compression. Distracted discs showed more BMP-2 (19.67 vs. 3.67 in 56 days compression) and collagen 2 (18.67 vs. 11.33 in 56 days compression) positive cells per field. Distraction results in disc rehydration, stimulated extracellular matrix gene expression, and increased numbers of protein-expressing cells.


The ADRP domain from a virulent strain of infectious bronchitis virus is not sufficient to confer a pathogenic phenotype to the attenuated Beaudette strain.
The replicase gene of the coronavirus infectious bronchitis virus (IBV) encodes 15 non-structural proteins (nsps). Nsp 3 is a multi-functional protein containing a conserved ADP-ribose-1″-phosphatase (ADRP) domain. The crystal structures of the domain from two strains of IBV, M41 (virulent) and Beaudette (avirulent), identified a key difference; M41 contains a conserved triple-glycine motif, whilst Beaudette contains a glycine-to-serine mutation that is predicted to abolish ADRP activity. Although ADRP activity has not been formally demonstrated for IBV nsp 3, Beaudette fails to bind ADP-ribose. The role of ADRP in virulence was investigated by generating rIBVs, based on Beaudette, containing either a restored triple-glycine motif or the complete M41 ADRP domain. Replication in vitro was unaffected by the ADRP modifications and the in vivo phenotype of the rIBVs was found to be apathogenic, indicating that restoration of the triple-glycine motif is not sufficient to restore virulence to the apathogenic Beaudette strain.


Multilocus analysis of SNP and metabolic data within a given pathway.
Complex traits, which are under the influence of multiple and possibly interacting genes, have become a subject of new statistical methodological research. One of the greatest challenges facing human geneticists is the identification and characterization of susceptibility genes for common multifactorial diseases and their association to different quantitative phenotypic traits. Two types of data from the same metabolic pathway were used in the analysis: categorical measurements of 18 SNPs; and quantitative measurements of plasma levels of several steroids and their precursors. Using the combinatorial partitioning method we tested various thresholds for each metabolic trait and each individual SNP locus. One SNP in CYP19, 3UTR, two SNPs in CYP1B1 (R48G and A119S) and one in CYP1A1 (T461N) were significantly differently distributed between the high and low level metabolic groups. The leave one out cross validation method showed that 6 SNPs in concert make 65% correct prediction of phenotype. Further we used pattern recognition, computing the p-value by Monte Carlo simulation to identify sets of SNPs and physiological characteristics such as age and weight that contribute to a given metabolic level. Since the SNPs detected by both methods reside either in the same gene (CYP1B1) or in 3 different genes in immediate vicinity on chromosome 15 (CYP19, CYP11 and CYP1A1) we investigated the possibility that they form intragenic and intergenic haplotypes, which may jointly account for a higher activity in the pathway. We identified such haplotypes associated with metabolic levels. The methods reported here may enable to study multiple low-penetrance genetic factors that together determine various quantitative phenotypic traits. Our preliminary data suggest that several genes coding for proteins involved in a common pathway, that happen to be located on common chromosomal areas and may form intragenic haplotypes, together account for a higher activity of the whole pathway.


The use of computed tomography attenuation to evaluate osteoporosis following acute fractures of the thoracic and lumbar vertebra.
Relatively few patients have dual-energy x-ray absorptiometry to quantify the magnitude of bone loss as they age. Recent work correlates mean computed tomography (CT) attenuation in the level I (L1) vertebra with bone mineral density (BMD), making it possible to objectively evaluate the magnitude of bone loss in osteoporosis by this method. The aims of this study were to evaluate the utility of using CT scans in patients with acute thoracic and lumbar spine fractures to diagnose osteoporosis and using CT attenuation to evaluate the association between age and BMD. We performed a retrospective study of patients with acute fractures of the thoracic or lumbar spine who had also undergone an abdominal (or L1) CT scan and compared mean CT attenuation in L1 against threshold values. We also compared differences in CT attenuation between younger (<65 years) and older (≥65 years) and older patients. A total of 124 patients were evaluated (74 thoracic and 50 lumbar fractures). Overall, there was a strong correlation between age and bone density as measured by CT attenuation (r = -.76). Among those with thoracic fractures (<65 years), mean CT attenuation was 196.51 HU. Forty-one patients were ≥65 years and had mean CT attenuation of 105.90 HU (P < .001). In patients with lumbar fractures, 27 patients were <65 years and had a mean CT attenuation of 192.26 HU and 23 patients were ≥65 years and had mean CT attenuation of 114.31 HU (P < .001). At the threshold of 110 HU, set for specificity, the magnitude of difference between the age-stratified cohorts was greater in the thoracic spine (P < .0001 vs P = .003). Using opportunistic CT, we demonstrate the relative frequency of osteoporosis in patients with acute fractures of the thoracic and lumbar spine and confirm that the association increases with age. The CT attenuation may provide a cheap and convenient method to help confirm a clinical diagnosis of osteoporosis in patients with fractures.


Big mountains but small barriers: population genetic structure of the Chinese wood frog (Rana chensinensis) in the Tsinling and Daba Mountain region of northern China.
Amphibians in general are poor dispersers and highly philopatric, and landscape features often have important impacts on their population genetic structure and dispersal patterns. Numerous studies have suggested that genetic differentiation among amphibian populations are particularly pronounced for populations separated by mountain ridges. The Tsinling Mountain range of northern China is a major mountain chain that forms the boundary between the Oriental and Palearctic zoogeographic realms. We studied the population structure of the Chinese wood frog (Rana chensinensis) to test whether the Tsinling Mountains and the nearby Daba Mountains impose major barriers to gene flow. Using 13 polymorphic microsatellite DNA loci, 523 individuals from 12 breeding sites with geographical distances ranging from 2.6 to 422.8 kilometers were examined. Substantial genetic diversity was detected at all sites with an average of 25.5 alleles per locus and an expected heterozygosity ranging from 0.504 to 0.855, and two peripheral populations revealed significantly lower genetic diversity than the central populations. In addition, the genetic differentiation among the central populations was statistically significant, with pairwise FST values ranging from 0.0175 to 0.1625 with an average of 0.0878. Furthermore, hierarchical AMOVA analysis attributed most genetic variation to the within-population component, and the between-population variation can largely be explained by isolation-by-distance. None of the putative barriers detected from genetic data coincided with the location of the Tsinling Mountains. The Tsinling and Daba Mountains revealed no significant impact on the population genetic structure of R. chensinensis. High population connectivity and extensive juvenile dispersal may account for the significant, but moderate differentiation between populations. Chinese wood frogs are able to use streams as breeding sites at high elevations, which may significantly contribute to the diminishing barrier effect of mountain ridges. Additionally, a significant decrease in genetic diversity in the peripheral populations supports Mayr's central-peripheral population hypothesis.


GMP synthetase stimulates histone H2B deubiquitylation by the epigenetic silencer USP7.
The packaging of eukaryotic genomic DNA into chromatin is modulated through a range of posttranslational histone modifications. Among these, the role of histone ubiquitylation remains poorly understood. Here, we show that the essential Drosophila ubiquitin-specific protease 7 (USP7) contributes to epigenetic silencing of homeotic genes by Polycomb (Pc). We purified USP7 from embryo nuclear extracts as a stable heteromeric complex with guanosine 5'-monophosphate synthetase (GMPS). The USP7-GMPS complex catalyzed the selective deubiquitylation of histone H2B, but not H2A. Biochemical assays confirmed the tight association between USP7 and GMPS in Drosophila embryo extracts. Similar to USP7, mutations in GMPS acted as enhancers of Pc in vivo. USP7 binding to GMPS was required for histone H2B deubiquitylation and strongly augmented deubiquitylation of the human tumor suppressor p53. Thus, GMPS can regulate the activity of a ubiquitin protease. Collectively, these results implicate a biosynthetic enzyme in chromatin control via ubiquitin regulation.


Ethical Issues in Neuromarketing: "I Consume, Therefore I am!".
Neuromarketing is a recent interdisciplinary field which crosses traditional boundaries between neuroscience, neuroeconomics and marketing research. Since this nascent field is primarily concerned with improving marketing strategies and promoting sales, there has been an increasing public aversion and protest against it. These protests can be exemplified by the reactions observed lately in Baylor School of Medicine and Emory University in the United States. The most recent attempt to stop ongoing neuromarketing research in France is also remarkable. The pertaining ethical issues have been continuously attracting much attention, especially since the number of neuromarketing companies has exceeded 300 world-wide. This paper begins with a brief introduction to the field of neurotechnology by presenting its current capabilities and limitations. Then, it will focus on the ethical issues and debates most related with the recent applications of this technology. The French Parliament's revision of rules on bioethics in 2004 has an exemplary role in our discussion. The proposal by Murphy et al. (2008) has attracted attention to the necessity of ethical codes structuring this field. A code has recently been declared by the Neuromarketing Science and Business Association. In this paper, it is argued that these technologies should be sufficiently discussed in public spheres and its use on humans should be fully carried out according to the ethical principles and legal regulations designed in line with human rights and human dignity. There is an urgent need in the interdisciplinary scientific bodies like ethics committees monitoring the research regarding the scientific and ethical values of nonmaleficence, beneficence, autonomy, confidentiality, right to privacy and protection of vulnerable groups.


Beyond artificial intelligence: exploring artificial wisdom.
The ultimate goal of artificial intelligence (AI) is to develop technologies that are best able to serve humanity. This will require advancements that go beyond the basic components of general intelligence. The term "intelligence" does not best represent the technological needs of advancing society, because it is "wisdom", rather than intelligence, that is associated with greater well-being, happiness, health, and perhaps even longevity of the individual and the society. Thus, the future need in technology is for artificial wisdom (AW). We examine the constructs of human intelligence and human wisdom in terms of their basic components, neurobiology, and relationship to aging, based on published empirical literature. We review the development of AI as inspired and driven by the model of human intelligence, and consider possible governing principles for AW that would enable humans to develop computers which can operationally utilize wise principles and result in wise acts. We review relevant examples of current efforts to develop such wise technologies. AW systems will be based on developmental models of the neurobiology of human wisdom. These AW systems need to be able to a) learn from experience and self-correct; b) exhibit compassionate, unbiased, and ethical behaviors; and c) discern human emotions and help the human users to regulate their emotions and make wise decisions. A close collaboration among computer scientists, neuroscientists, mental health experts, and ethicists is necessary for developing AW technologies, which will emulate the qualities of wise humans and thus serve the greatest benefit to humanity. Just as human intelligence and AI have helped further the understanding and usefulness of each other, human wisdom and AW can aid in promoting each other's growth.


Leading change: evidence-based transition.
The purpose of this article was to provide a framework for evidence-based transition of patient populations within an acute care pediatric institution. Transition within a hospital is foreseeable, given the ever-changing needs of the patients within an evolving healthcare system. These changes include moving patient populations because of expansion, renovation, or cohorting similar patient diagnoses to provide care across a continuum. Over the past 1 to 2 years, Children's Health Children's Medical Center Dallas has experienced a wide variety of transition. To provide a smooth transition for patients and families into new care areas resulting in a healthy work environment for all team members. The planning phase for patient population moves, and transition should address key aspects to include physical location and care flow, supplies and equipment, staffing model and human resources (HR), education and orientation, change process and integrating teams, and family preparation. It is imperative to consider these aspects in order for transitions within a healthcare system to be successful. During a time of such transitions, the clinical nurse specialist (CNS) is a highly valuable team member offering a unique perspective and methodological approach, which is central to the new initiative's overall success. The themes addressed in this article on evidence-based transition are organized according to the CNS spheres of influence: system/organization, patient/family, and nursing. An evidence-based transition plan was developed and implemented successfully with the support from the CNS for 3 patient populations. Organizational leadership gained an increased awareness of the CNS role at the conclusion of each successful transition. The CNS plays a pivotal role as clinical experts and proponents of evidence-based practice and effects change in the system/organization, nursing, and patient/family spheres of influence. While transitions can be a source of stress for leaders and bedside staff, it is also a time that allows for growth and new opportunities for staff and may result in development of a healthier work environment. The CNS is able to provide leadership while working collaboratively to oversee the moves with a forward-thinking approach. There are key components to consider during times of transition. These include (1) organize, plan, and improve work efficiencies during a construction build; (2) identify the key elements for improvement in nurse and patient satisfaction; (3) develop or maintain healthy work environment standards; (4) establish adequate staffing levels and staff education to successfully care for patient populations following transition; and (5) support the staff and patients during transition.


The tyrosine kinase c-Src enhances RIG-I (retinoic acid-inducible gene I)-elicited antiviral signaling.
Antiviral immune responses are initiated through Toll-like receptors (TLRs) and RIG-I (retinoic acid-inducible gene-I)-like RNA helicases that recognize nucleic acids from distinct viruses. In this study, we show that the tyrosine kinase c-Src participates in antiviral responses induced by the cytoplasmic RNA helicase RIG-I. Sendai virus (SV), which is recognized by RIG-I, induced c-Src phosphorylation. Functional impairment of c-Src through chemical inhibition or transient expression of a c-Src kinase-inactive mutant attenuated production of endogenous antiviral proteins after SV infection or after expression of RIG-I or its adapter protein MAVS. Importantly, SV-stimulated synthesis of antiviral proteins was significantly impaired in cells treated with c-Src small interfering RNA and in cells from c-Src-deficient mice. In addition, we found that c-Src interacted with components of the RIG-I pathway: RIG-I, MAVS, and TRAF3 (tumor necrosis factor receptor-associated factor-3). The interaction between c-Src and TRAF3 was found to occur within the RING domain of TRAF3. Taken together, our results suggest that c-Src enhances RIG-I-mediated signaling, acting at the level of TRAF3.


Rigidity coincides with reduced cognitive control to affective cues in children with autism.
The present study tested whether salient affective cues would negatively influence cognitive control in children with and without autism spectrum disorder (ASD). One hundred children aged 6-12 years who were either typically developing or had ASD performed a novel go/no-go task to cues of their interest versus cues of noninterest. Linear mixed-effects (LME) models for hit rate, false alarms, and the sensitivity index d' were used to test for group differences. Caregivers completed the Repetitive Behavior Scale-Revised to test associations between repetitive behaviors and task performance. Children with ASD had reduced cognitive control toward their interests compared with typically developing children. Further, children with ASD showed reduced cognitive control to interests compared with noninterests, a pattern not observed in typically developing children. Decreased cognitive control toward interests was associated with higher insistence on sameness behavior in ASD, but there was no association between sameness behavior and cognitive control for noninterests. Together, children with ASD demonstrated decreased cognitive flexibility in the context of increased affective salience related to interests. These results provide a mechanism for how salient affective cues, such as interests, interfere with daily functioning and social communication in ASD. Further, the findings have broader clinical implications for understanding how affective cues can drive interactions between restricted patterns of behavior and cognitive control. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Developing consensus on hospital prescribing indicators of potential harms amenable to decision support.
To develop a list of prescribing indicators specific for the hospital setting that would facilitate the prospective collection of high-severity and/or high-frequency prescribing errors, which are also amenable to electronic clinical decision support. A two-stage consensus technique (electronic Delphi) was carried out with 20 experts across England. Participants were asked to score prescribing errors using a five-point Likert scale for their likelihood of occurrence and the severity of the most likely outcome. These were combined to produce risk scores, from which median scores were calculated for each indicator across the participants in the study. The degree of consensus between the participants was defined as the proportion that gave a risk score in the same category as the median. Indicators were included if a consensus of 80% or more was achieved. A total of 80 prescribing errors were identified by consensus as being high or extreme risk. The most common drug classes named within the indicators were antibiotics (n = 13), antidepressants (n = 8), nonsteroidal anti-inflammatory drugs (n = 6) and opioid analgesics (n = 6). The most frequent error type identified as high or extreme risk were those classified as clinical contraindications (n = 29 of 80). Eighty high-risk prescribing errors in the hospital setting have been identified by an expert panel. These indicators can serve as a standardized, validated tool for the collection of prescribing data in both paper-based and electronic prescribing processes. This can assess the impact of safety improvement initiatives, such as the implementation of electronic clinical decision support.


[Combating the new influenza A (H1N1) virus. II. Surveillance and non-pharmaceutical interventions].
Intensive surveillance in Mexico, the US, Canada, Spain and the UK has resulted in the first data on the new influenza A (H1N1) virus. As of yet, it is still unknown if, how and where further transmission within the Netherlands will take place. Therefore, intensified surveillance in the Netherlands is essential. Information on individual patients and their contacts is available through the compulsory notification of the new influenza. The number of general practitioners and nursing homes participating in surveillance through national networks is extended. In addition mortality statistics and internet-based surveillance are being used (the Dutch Great Influenza Survey). Based on studies on previous flu epidemics and mathematical models we can estimate the expected effect of non-pharmaceutical interventions.The reduction of social contacts, in particular through the closure of schools, hand washing routines and wearing of facemasks if done correctly, are expected to be effective non-pharmaceutical interventions in slowing the transmission of the new influenza virus.


Anemia as the Main Manifestation of Myelodysplastic Syndromes.
Myelodysplastic syndromes (MDS) are a constellation of different diseases sharing anemia in the great majority of cases, and this cytopenia defines these pathologies and their most dramatic clinical manifestations. Anemia in MDS is due to ineffective erythropoiesis, with a high degree of apoptosis of marrow erythroid progenitors. These progenitors show distinctive dysplastic features that consent diagnosis, and are recognizable and differentiated, although not easily, from other morphologic alterations present in other types of anemia. Reaching the diagnosis of MDS in a macrocytic anemia and alleviating the symptoms of anemia are therefore an essential objective of the treating physician. In this work, the signs and symptoms of anemia in MDS, as well as its peculiar pathophysiology, are discussed. Erythopoietic stimulating agents (ESAs) are providing the best treatment for anemic MDS patients, but their use is still not approved by health agencies. While still waiting for this waiver, their clinical use is widespread and their effectivness is well known, as well as the dismal prognosis of patients who do not respond to ESAs and require transfusions. MDS with del5q constitute a unique model of anemia whose complex pathophysiology has been clarified at least partially, defining its link to ribosomal alterations likewise what observed in hereditary anemias like Blackfan Diamond anemia. Lenalidomide is the agent that has shown striking and specific erythropoietic activity in del5q MDS, and the basis of this response is starting to be understood. Several new agents are under evaluation for ESA refractory/relapsed MDS patients, targeting different putative mechanisms of ineffective erythropoiesis, and are here reviewed.


Testing Rare-Variant Association without Calling Genotypes Allows for Systematic Differences in Sequencing between Cases and Controls.
Next-generation sequencing of DNA provides an unprecedented opportunity to discover rare genetic variants associated with complex diseases and traits. However, the common practice of first calling underlying genotypes and then treating the called values as known is prone to false positive findings, especially when genotyping errors are systematically different between cases and controls. This happens whenever cases and controls are sequenced at different depths, on different platforms, or in different batches. In this article, we provide a likelihood-based approach to testing rare variant associations that directly models sequencing reads without calling genotypes. We consider the (weighted) burden test statistic, which is the (weighted) sum of the score statistic for assessing effects of individual variants on the trait of interest. Because variant locations are unknown, we develop a simple, computationally efficient screening algorithm to estimate the loci that are variants. Because our burden statistic may not have mean zero after screening, we develop a novel bootstrap procedure for assessing the significance of the burden statistic. We demonstrate through extensive simulation studies that the proposed tests are robust to a wide range of differential sequencing qualities between cases and controls, and are at least as powerful as the standard genotype calling approach when the latter controls type I error. An application to the UK10K data reveals novel rare variants in gene BTBD18 associated with childhood onset obesity. The relevant software is freely available.


Ethicist as designer: a pragmatic approach to ethics in the lab.
Contemporary literature investigating the significant impact of technology on our lives leads many to conclude that ethics must be a part of the discussion at an earlier stage in the design process i.e., before a commercial product is developed and introduced. The problem, however, is the question regarding how ethics can be incorporated into an earlier stage of technological development and it is this question that we argue has not yet been answered adequately. There is no consensus amongst scholars as to the kind of ethics that should be practiced, nor the individual selected to perform this ethical analysis. One school of thought holds that ethics should have pragmatic value in research and design and that it should be implemented by the (computer) engineers and/or (computer) scientists themselves, while another school of thought holds that ethics need not be so pragmatic. For the latter, the ethical reflection can aim at a variety of goals, and be carried out by an ethicist. None of the approaches resulting from these lines of thinking have been adopted on a wide-scale basis. To that end, the approach presented here is intended to bridge the gap between these schools of thought. It is our contention that ethics ought to be pragmatic and to provide utility for the design process and we maintain that adequate ethical reflection, and all that it entails, ought to be conducted by an ethicist. Thus, we propose a novel role for the ethicist--the ethicist as designer--who subscribes to a pragmatic view of ethics in order to bring ethics into the research and design of artifacts-no matter the stage of development. In this paper we outline the series of steps that a pragmatic value analysis entails: uncovering relevant values, scrutinizing these values and, working towards the translation of values into technical content. In conclusion, we present a list of tasks for the ethicist in his/her role as designer on the interdisciplinary team.


Activation of adenosine A(3) receptors potentiates stimulatory effects of IL-3, SCF, and GM-CSF on mouse granulocyte-macrophage hematopoietic progenitor cells.
Adenosine A(3) receptor agonist N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) has been tested from the point of view of potentiating the effects of hematopoietic growth factors interleukin-3 (IL-3), stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) on the growth of hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in suspension of normal mouse bone marrow cells in vitro. IB-MECA alone induced no GM-CFC growth. Significant elevation of numbers of GM-CFC evoked by the combinations of IB-MECA with IL-3, SCF, or GM-CSF as compared with these growth factors alone has been noted. Combination of IB-MECA with G-CSF did not induce significantly higher numbers of GM-CFC in comparison with G-CSF alone. Joint action of three drugs, namely of IB-MECA + IL-3 + GM-CSF, produced significantly higher numbers of GM-CFC in comparison with the combinations of IB-MECA + IL-3, IB-MECA + GM-CSF, or IL-3 + GM-CSF. These results give evidence of a significant role of selective activation of adenosine A(3) receptors in stimulation of the growth of granulocyte/ macrophage hematopoietic progenitor cells.


Occipital and orbitofrontal hemodynamics during naturally paced reading: an fNIRS study.
Humans typically read at incredibly fast rates, because they predict likely occurring words from a given context. Here, we used functional near-infrared spectroscopy (fNIRS) to track the ultra-rapid hemodynamic responses of words presented every 280 ms in a naturally paced sentence context. We found a lower occipital deoxygenation to unpredictable than to predictable words. The greater hemodynamic responses to unexpected words suggest that the visual features of expected words have been pre-activated previous to stimulus presentation. Second, we tested opposing theoretical proposals about the role of the medial orbitofrontal cortex (OFC): Either OFC may respond to the breach of expectation; or OFC is activated when the present stimulus matches the prediction. A significant interaction between word frequency and predictability indicated OFC responses to breaches of expectation for low- but not for high-frequency words: OFC is sensitive to both, bottom-up processing as mediated by word frequency, as well as top-down predictions. Particularly, when a rare word is unpredictable, OFC becomes active. Finally, we discuss how a high temporal resolution can help future studies to disentangle the hemodynamic responses of single trials in such an ultra-rapid event succession as naturally paced reading.


Early differentiation and migration of cranial neural crest in the opossum, Monodelphis domestica.
Marsupial mammals are born at a highly altricial state. Nonetheless, the neonate must be capable of considerable functional independence. Comparative studies have shown that in marsupials the morphogenesis of many structures critical to independent function are advanced relative to overall development. Many skeletal and muscular elements in the facial region show particular heterochrony. Because neural crest cells are crucial to forming and patterning much of the face, this study investigates whether the timing of cranial neural crest differentiation is also advanced. Histology and scanning electron microscopy of Monodelphis domestica embryos show that many aspects of cranial neural crest differentiation and migration are conserved in marsupials. For example, as in other vertebrates, cranial neural crest differentiates at the neural ectoderm/epidermal boundary and migrates as three major streams. However, when compared with other vertebrates, a number of timing differences exist. The onset of cranial neural crest migration is early relative to both neural tube development and somite formation in Monodelphis. First arch neural crest cell migration is particularly advanced and begins before any somites appear or regional differentiation exists in the neural tube. Our study provides the first published description of cranial neural crest differentiation and migration in marsupials and offers insight into how shifts in early developmental processes can lead to morphological change.


Age-associated differences in cardiovascular inflammatory gene induction during endotoxic stress.
Upon physiological stress, families of stress response genes are activated as natural defense mechanisms. Here, we show that induction of specific inflammatory genes is significantly dysregulated and altered in the heart of aged (24--26-month-old) versus young (4-month-old) mice experimentally challenged with a bacterial endotoxin, lipopolysaccharide (LPS, 1.5 mg/kg of body mass). Whereas the LPS-mediated induction of cardiac mRNA for tumor necrosis factor alpha or inducible nitric-oxide synthase showed no age-associated differences, the induction of interleukin-1 beta (IL-1 beta) and intracellular adhesion molecule-1 was modestly extended with aging, and the induction of IL-6 was significantly prolonged with aging. This age-associated phenomenon occurred gradually from 4 to 17 months of age and became more evident after 23 months of age. The age-associated augmentation of the cardiac IL-6 induction was also dramatic at the protein level. Immunohistochemically, the LPS-induced cardiac IL-6 was localized mainly in the microvascular walls. Aged but not young mice showed a high mortality rate during these experiments. These results demonstrate that endotoxin-mediated induction of specific inflammatory genes in cardiovascular tissues is altered with aging, which may be causally related to the increased susceptibility of aged animals to endotoxic stress.