Increased incidence of fractures in middle-aged and elderly men with low intakes of phosphorus and zinc.
The aim of the study was to determine dietary risk factors for fracture in men aged 46-68 years. Six thousand five hundred and seventy-six men were randomly invited using the Municipal Registry to a diet and health study. The diet was assessed using a combined 7-day menu book for hot meals, beverages and dietary supplements and a quantitative food frequency questionnaire for other foods. The fracture incidence was 103/10,000 person-years during a mean follow-up of 2.4 years. Zinc and phosphorus intake were associated with fracture risk and showed a threshold effect. The zinc intake in the lowest decentile, 10 mg daily, was associated with almost a doubled risk of fracture compared with the fourth and fifth quintiles (RR = 0.47; 95% confidence interval, 27-82) of zinc intake adjusted for energy, previous fractures, lifestyle factors and co-morbidity. Energy-adjusted phosphorus intake in the lowest quintile, mean level 1357 mg, was associated with an increased fracture risk compared with subjects in the second quintile. Smoking, martial status and physical activity were independently associated with fracture risk. Calcium, retinol and vitamin D showed no associations with fracture risk. We conclude that inadequate intakes of zinc and phosphorus are important risk factors for fracture.


Prostate cancer: more effective use of underutilized postoperative radiation therapy.
Introduction: Adverse pathological features at radical prostatectomy such as extracapsular extension, seminal-vesicle involvement, positive surgical margins and/or lymph node invasion define a particular subgroup of patients that might benefit from additional treatment after surgery, in particular radiation therapy.Areas covered: Post-prostatectomy radiation is intended as adjuvant, early-salvage or salvage depending on the timing and PSA levels at the treatment. After providing the most used definitions, the high-level evidence supporting adjuvant radiation is reviewed together with the limitations affecting its utilization. In recent years early-salvage radiation was hypothesized to be a non-inferior alternative based on good-quality retrospective data. Recently, preliminary results of ongoing trials provide additional evidence. In light of the need to identify patients that will truly benefit from adjuvant radiation, clinically based and molecular tools available for this purpose are reviewed.Expert opinion: In order to tailor treatment for the patient after radical prostatectomy, there is a need for a tool that could both improve the oncological outcomes and be cost-effective. To date, genomic testing provides the most promising results that will be reasonably improved in the coming years.


Morphological Correlates of KIT and PDGFRA Genotypes in Gastrointestinal Stromal Tumour.
The aim of the study was to study the clinicopathological and immunohistochemical features of gastrointestinal stromal tumours and correlation with KIT/PDGFRA mutations. Eighty consecutive resected cases were genotyped for KIT exons 11, 9, 13, 17 and PDGFRA exons 18, 14, 12 and correlated with histomorphology by nonparametric tests. Forty-seven cases (58.8%) were in the high-risk group. Males had higher rates of KIT exon 11 and PDGFRA exon 18 mutations than females (p=0.03). KIT and PDGFRA mutation frequencies were lower (58.8%) than western data showing KIT exon 11 mutation in 63.8%, KIT exon 9 mutation in 19% and PDGFRA exon 18 mutation in 17% of the cases. Extragastrointestinal stromal tumours (n=6) showed 100% mutation. KIT exon 11 deletion was associated with gastric location (60%) (p=0.04), spindle cells (63.3%), and high-risk stratification (66.6%) (p=0.01) while KIT exon 9 mutation was common in small intestine (66.7%) (p=0.04), in higher risk groups (66.7%) (p=0.01) and 75% of codon 502-503 duplications (p=0.03). PDGFRA 18 mutation was common in males (p=0.03), in gastric location (62.5%) (p=0.04), in cases showing mild to moderate atypia (62.5%) (p=0.01) and lower risk stratification (62.5%) (p=0.01). KIT/PDGFRA mutations were significantly associated with gender (p=0.03), location (p=0.04), nuclear atypia (p=0.01) and risk stratification (p=0.01). Morphological features and anatomic location may be useful in deciding molecular testing strategy, particularly in resource-limited settings, when a plethora of targetable mutations are present. An algorithm may be derived for genotyping with KIT exon 11 and PDGFRA exon 18 heading the list of targetable mutations. This approach may reduce financial burden on patients as well as workload on hospital staff.


Cell adhesion molecule-1 (CADM1) expressed on adult T-cell leukemia/lymphoma cells is not involved in the interaction with macrophages.
Cell adhesion molecule 1 (CADM1) is a cell adhesion molecule that is expressed in brain, liver, lung, testis, and some kinds of cancer cells including adult T-cell leukemia/lymphoma (ATLL). Recent studies have indicated the involvement of CADM1 in cell-cell contact between cytotoxic T-lymphocytes and virus infected cells. We previously reported that cell-cell interaction between lymphoma cells and macrophages induces lymphoma cell proliferation. In the present study, we investigated whether CADM1 is associated with cell-cell interaction between several human lymphoma cell lines and macrophages.CADM1 expression was observed in the ATLL cell lines, ATN-1, ATL-T, and ATL-35T, and in the B cell lymphoma cell lines, TL-1, DAUDI, and SLVL, using western blotting. Significant cell-cell interaction between macrophages and ATN-1, ATL-T, ATL-35T and MT-2, DAUDI, and SLVL cells, as assessed by induction of cell proliferation, was observed. Immunohistochemical analysis of human biopsy samples indicated CADM1 expression in 10 of 14 ATLL cases; however, no case of follicular lymphoma or diffuse large B-cell lymphoma was positive for CADM1. Finally, the interaction of macrophages with cells of the CADM1-negative ED ATLL cell line and CADM1-transfected ED cells was tested. However, significant cell-cell interaction between macrophage and CADM1-transfected ED cells was not observed. We conclude that CADM1 was not associated with cell-cell interaction between lymphoma cells and macrophages, although CADM1 may be a useful marker of ATLL for diagnostic procedures.


Renal sympathetic nerve discharge mediated by the paraventricular nucleus is altered in STZ induced diabetic rats.
The purpose of this study was to determine if the renal sympathetic nerve discharge (RSND), mediated by the paraventricular nucleus of the hypothalamus (PVN), is altered in streptozotocin (STZ) induced diabetic rats. Two to three weeks prior to the start of the experiment the diabetic group was treated with a single injection of STZ (65 mg/kg ip) in a 2% solution of cold 0.1 M citrate buffer (pH = 4.5). The control group was injected with the vehicle alone. Bicuculline (BIC) was injected into the PVN in three different doses (0.5, 1.0 & 2.0 nmol) while the RSND was being monitored. BIC is an antagonist of gamma-aminobutyric acid (GABA) which is an inhibitory neurotransmitter that is thought to exert a tonic inhibitory effect on the PVN. Microinjection of BIC produced a significantly lower response of RSND in the diabetic group compared to the control group. Microinjection of BIC produced a 13, 35 and 33% change in RSND in diabetic rats compared to control group in response to 0.5, 1.0, and 2.0 nmol doses, respectively. Mean blood pressure and heart rate were statistically not different between the control and the diabetic groups. However there were tendencies of the blood pressure and heart rate to be blunted upon injection of BIC into PVN in diabetic rats. This study demonstrates that RSND in response to microinjection of BIC in PVN is decreased significantly in anesthetized diabetic rats, indicating that STZ induced diabetic rats have a blunted RSND response to microinjection of BIC. There is also some evidence, although not statistically significant, for a blunted blood pressure and heart rate were diabetics. These findings suggest that there is a reduced endogenous inhibitory influence of GABAergic mechanisms within the PVN involved in regulating renal sympathetic outflow in the diabetic state.


Self-control depletion and nicotine deprivation as precipitants of smoking cessation failure: A human laboratory model.
The need to understand potential precipitants of smoking relapse is exemplified by relapse rates as high as 95%. The Self-Control Strength model, which proposes that self-control is dependent upon limited resources and susceptible to fatigue, may offer insight into relapse processes. The current study tested the hypothesis that self-control depletion (SCD), produced from engagement in emotional suppression, would serve as a novel antecedent for cessation failure, as indexed by a validated laboratory analogue of smoking lapse and relapse. We also examined whether SCD effects interacted with those of a well-established relapse precipitant (i.e., nicotine deprivation). Craving and behavioral economic indices (delay discounting and demand) were tested as hypothesized mechanisms for increased cessation failure. Ultimately, a moderated mediation model was used to test nicotine deprivation as a hypothesized moderator of SCD effects. We used a 2 × 2 (12-hr deprivation vs. no deprivation; SCD vs. no SCD) factorial between-subjects design (N = 128 smokers). The primary hypothesis of the study was supported, as SCD increased lapse behavior (p = .04). Nicotine deprivation significantly increased craving, cigarette demand, delay discounting, and lapse behavior. No main effects were found for SCD on putative mediators (i.e., craving, demand, and discounting), but the SCD and deprivation manipulations interacted upon craving (p = .04). The moderated mediation model was significant. SCD was found to increase craving among nicotine deprived smokers, which mediated effects on lapse behavior. SCD appears to play an important role in smoking relapse and may be a viable target for intervention. (PsycINFO Database Record


Rattling the cage and opening the door.
I share with Poulin-Dubois and with Sabbagh, Koenig and Kuhlmeier the conviction that more research is needed on the mechanisms supporting selective social learning in infants and children. However, my plea is more specific: for research that tests domain-specific hypotheses about mechanism against domain-general hypotheses derived from other fields of cognitive science. Many, but not all, of these alternative hypotheses relate to domain-general mechanisms of attention.


Design of a Single-Layer Microchannel for Continuous Sheathless Single-Stream Particle Inertial Focusing.
High-throughput, high-precision single-stream focusing of microparticles has a potentially wide range of applications in biochemical analysis and clinical diagnosis. In this work, we develop a sheathless three-dimensional (3D) particle-focusing method in a single-layer microchannel. This novel microchannel consists of periodic high-aspect-ratio curved channels and straight channels. The proposed method takes advantage of both the curved channels, which induce Dean flow to promote particle migration, and straight channels, which suppress the remaining stirring effects of Dean flow to stabilize the achieved particle focusing. The 3D particle focusing is demonstrated experimentally, and the mechanism is analyzed theoretically. The effects of flow rate, particle size, and cycle number on the focusing performance were also investigated. The experimental results demonstrate that polystyrene particles with diameters of 5-20 μm can be focused into a 3D single file within seven channel cycles, with the focusing accuracy up to 98.5% and focusing rate up to 98.97%. The focusing throughput could reach up to ∼105 counts/min. Furthermore, its applicability to biological cells is also demonstrated by 3D focusing of HeLa and melanoma cells and bovine blood cells in the proposed microchannel. The proposed sheathless passive focusing scheme, featuring a simple channel structure, small footprint (9 mm × 1.2 mm), compact layout, and uncomplicated fabrication procedure, holds great promise as an efficient 3D focusing unit for the development of next-generation on-chip flow cytometry.


Combined use of Paracoccidioides brasiliensis recombinant 27-kilodalton and purified 87-kilodalton antigens in an enzyme-linked immunosorbent assay for serodiagnosis of paracoccidioidomycosis.
The diagnosis of paracoccidioidomycosis (PCM) has relied on the identification of the host's humoral response by using a variety of immunological methods, such as complement fixation and immunodiffusion. Although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. The use of combinations of purified, well-characterized antigens appears preferable and may yield optimum results. Accordingly an indirect enzyme-linked immunosorbent assay (ELISA) using combinations of the previously described 27-kDa recombinant antigen and the 87-kDa heat shock protein were used for diagnosis and follow-up of patients with PCM. A total of 37 patients classified according to their clinical presentations (7 with the acute or subacute form of the disease, 22 with the chronic form of the disease, and 8 with the chronic unifocal form) were studied. Eighteen of these patients were also evaluated at every follow-up appointment. Forty serum samples from patients with other diseases and 50 serum samples from healthy individuals were also studied. Detection of anti-27-kDa and anti-87-kDa antibodies in sera of patients with PCM by ELISA using a combination of the two purified proteins showed a sensitivity of 92% with a specificity of 88% in comparison with normal human sera and 90% in comparison with the heterologous sera. These results demonstrated a significant increase in sensitivity and specificity compared to results when the antigens were used separately. Thus, the use of combinations of well-defined antigens appears to offer clear advantages over the use of single antigens when diagnosing PCM.


Motion correction in optical coherence tomography volumes on a per A-scan basis using orthogonal scan patterns.
High speed Optical Coherence Tomography (OCT) has made it possible to rapidly capture densely sampled 3D volume data. One key application is the acquisition of high quality in vivo volumetric data sets of the human retina. Since the volume is acquired in a few seconds, eye movement during the scan process leads to distortion, which limits the accuracy of quantitative measurements using 3D OCT data. In this paper, we present a novel software based method to correct motion artifacts in OCT raster scans. Motion compensation is performed retrospectively using image registration algorithms on the OCT data sets themselves. Multiple, successively acquired volume scans with orthogonal fast scan directions are registered retrospectively in order to estimate and correct eye motion. Registration is performed by optimizing a large scale numerical problem as given by a global objective function using one dense displacement field for each input volume and special regularization based on the time structure of the acquisition process. After optimization, each volume is undistorted and a single merged volume is constructed that has superior signal quality compared to the input volumes. Experiments were performed using 3D OCT data from the macula and optic nerve head acquired with a high-speed ultra-high resolution 850 nm spectral OCT as well as wide field data acquired with a 1050 nm swept source OCT instrument. Evaluation of registration performance and result stability as well as visual inspection shows that the algorithm can correct for motion in all three dimensions and on a per A-scan basis. Corrected volumes do not show visible motion artifacts. In addition, merging multiple motion corrected and registered volumes leads to improved signal quality. These results demonstrate that motion correction and merging improves image quality and should also improve morphometric measurement accuracy from volumetric OCT data.


Acid has antiproliferative effects in nonneoplastic Barrett's epithelial cells.
For patients with Barrett's esophagus, physicians commonly prescribe antisecretory medications in dosages above those required to heal reflux esophagitis because acid has been shown to have proproliferative and antiapoptotic effects on Barrett's cancer cells and on Barrett's mucosal explants. For a number of reasons, these model systems may not be ideal for determining the effects of acid on benign Barrett's epithelial cells, however. We studied the effects of acid on proliferation and apoptosis in a nonneoplastic, telomerase-immortalized Barrett's epithelial cell line. Barrett's cells were treated with two 3-minute exposures to acidic media. Cell growth was determined using cell counts, proliferation was studied by flow cytometry, cell viability was determined by trypan blue staining, and apoptosis was assessed by TUNEL and Annexin V. The expression levels of p53 and p21 were determined by Western blotting. p53 siRNA was used to study the effect of p53 inhibition on total cell numbers after acid exposure. Acid exposure significantly decreased total cell numbers at 24 h without affecting either cell viability or apoptosis. Acid exposure resulted in cell cycle prolongation that was associated with greater expression of p53, but not p21. The acid-induced decrease in total cell numbers was abolished by p53 RNAi. Acid exposure has p53-mediated, antiproliferative effects in nonneoplastic Barrett's epithelial cells. These findings contradict the results of prior in vitro and ex vivo studies. We speculate that the prescription of antisecretory medications in dosages beyond those required to heal gastroesophageal reflux disease (GERD) symptoms and endoscopic signs could be detrimental. Controlled, prospective clinical trials are needed to determine the optimal level of acid suppression for patients with Barrett's esophagus.


Multilineage differentiation from human embryonic stem cell lines.
Stem cells are unique cell populations with the ability to undergo both self-renewal and differentiation. A wide variety of adult mammalian tissues harbors stem cells, yet "adult" stem cells may be capable of developing into only a limited number of cell types. In contrast, embryonic stem (ES) cells, derived from blastocyst-stage early mammalian embryos, have the ability to form any fully differentiated cell of the body. Human ES cells have a normal karyotype, maintain high telomerase activity, and exhibit remarkable long-term proliferative potential, providing the possibility for unlimited expansion in culture. Furthermore, they can differentiate into derivatives of all three embryonic germ layers when transferred to an in vivo environment. Data are now emerging that demonstrate human ES cells can initiate lineage-specific differentiation programs of many tissue and cell types in vitro. Based on this property, it is likely that human ES cells will provide a useful differentiation culture system to study the mechanisms underlying many facets of human development. Because they have the dual ability to proliferate indefinitely and differentiate into multiple tissue types, human ES cells could potentially provide an unlimited supply of tissue for human transplantation. Though human ES cell-based transplantation therapy holds great promise to successfully treat a variety of diseases (e.g., Parkinson's disease, diabetes, and heart failure) many barriers remain in the way of successful clinical trials.


Implementation of point-of-care testing in a pediatric healthcare setting.
Point-of-care testing (POCT) refers to testing performed outside the clinical laboratory near the patient or at the site of patient care. This could be in critical care settings like the intensive care unit (ICU) and emergency department (ED) or primary care settings like physician offices where testing is performed by nonlaboratory personnel. POCT circumvents several steps in central laboratory testing including specimen transportation and processing resulting in faster turnaround times. Provider access to rapid test results at the site of patient care allows for prompt medical decision making which can lead to improved patient outcomes, operational efficiencies, patient satisfaction, and even cost savings in some cases. In addition to providing results rapidly, POCT devices have small specimen volume requirements compared to central laboratory tests making POCT particularly attractive for pediatric healthcare settings. The availability of published reports on the impact of POCT implementation in pediatric care are helpful resources when evaluating the clinical necessity of POCT prior to implementation. Even though several studies have shown advantages to implementing POCT in different pediatric settings, it is important to note that limitations exist that might limit the utilization of certain POCTs in some pediatric populations. So, it is important that these limitations and the analytical performance of a test are considered while keeping the target patient population in mind. Since POCTs are performed by non-laboratory staff who are not trained laboratory personnel, one challenge with POCT is maintaining regulatory compliance and quality assurance. It is therefore important that regulatory and quality assurance programs be put in place prior to implementing POCT in the pediatric hospital. With advances in POCT technology, most POCT devices have the capability to interface to the laboratory information system (LIS) and electronic medical record (EMR). POCT device interfacing allows for improved compliance to regulatory and quality assurance standards. Maintaining a cost efficient POCT program is becoming increasingly important as hospitals and healthcare systems are undergoing consolidation and harmonization. This includes assessing the clinical and operational benefit of POCT before implementation and inventory management to ensure minimal reagent wastage. This review discusses these different considerations when implementing POCT with a focus on the pediatric healthcare setting.


Hilbert phase microscopy for investigating fast dynamics in transparent systems.
We introduce Hilbert phase microscopy (HPM) as a novel optical technique for measuring high transverse resolution quantitative phase images associated with optically transparent objects. Because of its single-shot nature, HPM is suitable for investigating rapid phenomena that take place in transparent structures such as biological cells. The potential of this technique for studying biological systems is demonstrated with measurements of red blood cells, and its ability to quantify dynamic processes on a millisecond scale is exemplified with measurements of evaporating micrometer-sized water droplets.


Neurochemical and behavioral alterations elicited by a chronic intermittent stressor regimen: implications for allostatic load.
Although stressors induce a series of adaptive neurochemical changes, sustained physiological activation associated with protracted stressor exposure may engender adverse effects (allostatic load). In the present investigation CD-1 mice exposed to a series of different stressors, twice a day over 54 days, exhibited increased signs of depression and anxiety, including increased passivity in a forced swim test, reduced aggression in a social interaction test, and delayed approach to food in a novel environment. Consistent with the view that a chronic stressor regimen affects immune-related processes, sickness behavior elicited by the proinflammatory cytokine, interleukin-1beta, was augmented in response to a chronic but not an acute stressor. Relative to nonstressed mice, median eminence serotonin was augmented by the cytokine treatment administered 24 h after chronic stressor exposure. Treatment with IL-1beta diminished plasma growth hormone levels and increased circulating corticosterone levels irrespective of the animals stressor history. It is suggested that chronic stressor exposure may instigate relatively protracted neurochemical effects, thereby influencing the behavioral responses to later psychological and systemic challenges.


NANOmetric BIO-Banked MSC-Derived Exosome (NANOBIOME) as a Novel Approach to Regenerative Medicine.
Mesenchymal stem cells (MSCs) are well known for their great potential in clinical applications. In fact, MSCs can differentiate into several cell lineages and show paracrine behavior by releasing endogenous factors that stimulate tissue repair and modulate local immune response. Each MSC type is affected by specific biobanking issues-technical issues as well as regulatory and ethical concerns-thus making it quite tricky to safely and commonly use MSC banking for swift regenerative applications. Extracellular vesicles (EVs) include a group of 150⁻1000 nm vesicles that are released by budding from the plasma membrane into biological fluids and/or in the culture medium from varied and heterogenic cell types. EVs consist of various vesicle types that are defined with different nomenclature such as exosomes, shedding vesicles, nanoparticles, microvesicles and apoptotic bodies. Ectosomes, micro- and nanoparticles generally refer to the direct release of single vesicles from the plasma membrane. While many studies describe exosomes as deriving from multivesicular bodies, solid evidence about the origin of EVs is often lacking. Extracellular vesicles represent an important portion of the cell secretome. Their numerous properties can be used for diagnostic, prognostic, and therapeutic uses, so EVs are considered to be innovative and smart theranostic tools. The aim of this review is to investigate the usefulness of exosomes as carriers of the whole information panel characterizing the use of MSCs in regenerative medicine. Our purpose is to make a step forward in the development of the NANOmetric BIO-banked MSC-derived Exosome (NANOBIOME).


Neonatal, infant and under-five mortalities in Nigeria: An examination of trends and drivers (2003-2013).
Neonatal (NMR), infant (IMR) and under-five (U5M) mortality rates remain high in Nigeria. Evidence-based knowledge of trends and drivers of child mortality will aid proper interventions needed to combat the menace. Therefore, this study assessed the trends and drivers of NMR, IMR, and U5M over a decade in Nigeria. A nationally representative data from three consecutive Nigeria Demographic and Household Surveys (NDHS) was used. A total of 66,158 live births within the five years preceding the 2003 (6029), 2008 (28647) and 2013 (31482) NDHS were included in the analyses. NMR was computed using proportions while IMR and U5 were computed using life table techniques embedded in Stata version 12. Probit regression model and its associated marginal effects were used to identify the predisposing factors to NMR, IMR, and U5M. The NMR, IMR, and U5M per 1000 live births in 2003, 2008 and 2013 were 52, 41, 39; 100, 75, 69; and 201, 157, 128 respectively. The NMR, IMR, and U5M were consistently lower among children whose mothers were younger, living in rural areas and from richer households. Generally, the probability of neonate death in 2003, 2008 and 2013 were 0.049, 0.039 and 0.038 respectively, the probability of infant death was 0.093, 0.071 and 0.064 while the probability of under-five death was 0.140, 0.112 and 0.092 for the respective survey years. While adjusting for other variables, the likelihood of infant and under-five deaths was significantly reduced across the survey years. Maternal age, mothers' education, place of residence, child's sex, birth interval, weight at birth, skill of birth attendant, delivery by caesarean operation or not significantly influenced NMR, IMR, and U5M. The NMR, IMR, and U5M in Nigeria reduced over the studied period. Multi-sectoral interventions targeted towards the identified drivers should be instituted to improve child survival.


Pharmacologic treatment of anorexia nervosa: where do we go from here?
Anorexia nervosa (AN) is a serious mental disorder, characterized by severely low weight and cognitive distortions about body shape and weight. AN is generally associated with a range of psychological symptoms, including depression, anxiety, obsessions, and rituals. The current study summarized findings from randomized controlled trials (RCT) using pharmacologic treatments in patients with AN. We conducted a review of literature using Medline. Several classes of pharmacologic agents have been studied in small samples of patients with acute AN without finding clear benefit to eating, weight, body shape concerns, or associated psychopathology. Studies have been limited by small sample sizes, as well as by research design with most studies adding medication to comprehensive hospital-based treatment programs. Future directions for pharmacologic treatment research in AN should include outpatient trials, rigorous study of atypical antipsychotic medication, and assessment of medication effect for relapse prevention in weight-restored patients.


Miltefosine in the treatment of leishmaniasis: Clinical evidence for informed clinical risk management.
Visceral leishmaniasis (VL) is a life-threatening disease. Traditional treatment with pentavalent antimony injections has become ineffective in the area with the world's highest prevalence of disease (North Bihar, India) and is becoming less effective elsewhere as well. A replacement is needed, best if it can be given to more patients outside the hospital. Miltefosine is the first oral drug registered for VL. Given daily under medical supervision for 4 weeks, it cures 94% of patients (both children and adults) and is reasonably safe. Miltefosine has great potential for improving access to treatment and overall control of VL and will be critical in the VL elimination campaign in the Indian subcontinent, but must be safeguarded or will be lost if misused. Its main limitations are adherence (and hence potential for selection of drug resistant parasites) and teratogenicity (pregnancy must be avoided during treatment and the following two months). This calls for responsible deployment, setting in place mechanisms to protect female patients in child-bearing age, monitoring effects and optimizing adherence in real-life conditions through directly observed therapy. One option to protect the useful life-span of miltefosine consists in shortening treatment duration by combining it with another drug.


A systematic review of antimicrobial resistance in Salmonella enterica serovar Typhi, the etiological agent of typhoid.
The temporal and spatial change in trends of antimicrobial resistance (AMR) in typhoid have not been systematically studied, and such information will be critical for defining intervention, as well as planning sustainable prevention strategies. To identify the phenotypic trends in AMR, 13,833 individual S. Typhi isolates, reported from 1973 to 2018 in 62 publications, were analysed to determine the AMR preponderance over time. Separate analyses of molecular resistance determinants present in over 4,000 isolates reported in 61 publications were also conducted. Multi-drug resistant (MDR) typhoid is in decline in Asia in a setting of high fluoroquinolone resistance while it is on the increase in Africa. Mutations in QRDRs in gyrA (S83F, D87N) and parC (S80I) are the most common mechanisms responsible for fluoroquinolone resistance. Cephalosporin resistant S. Typhi, dubbed extensively drug-resistant (XDR) is a real threat and underscores the urgency in deploying the Vi-conjugate vaccines. From these observations, it appears that AMR in S. Typhi will continue to emerge leading to treatment failure, changes in antimicrobial policy and further resistance developing in S. Typhi isolates and other Gram-negative bacteria in endemic regions. The deployment of typhoid conjugate vaccines to control the disease in endemic regions may be the best defence.


Resection of an Optic Canal Meningioma through a Contralateral Subfrontal Approach with Endoscopic Assistance: A 2D Operative Video.
Objective To review the use of the contralateral subfrontal approach for the resection of an optic canal meningioma. Design Operative video. Results A meningioma, located in the inferomedial side of the optic canal ( Fig. 1 ), was found to cause significant visual deterioration. The subfrontal route was preferred to expose the tumor without mobilization of the optic nerve. Drilling of the anterior limb of the chiasmatic sulcus (limbus sphenoidale) provided adequate exposure of the medial aspect of the optic canal. Gross total resection (Simpson II) of the tumor was accomplished, and endoscopic assistance allowed identification and coagulation of an infiltrated dura mater in the chiasmatic sulcus and tuberculum sellae. Conclusion The subfrontal approach grants an optimal surgical trajectory to the contralateral chiasmatic sulcus and optic nerve. When the medial side of the optic canal is drilled, tumors extending into the optic canal can be safely resected, under direct visualization of the inferomedial side of the optic nerve. Breach into the sphenoid sinus can occur during drilling of the anterior limb of the chiasmatic sulcus. Endoscopic assistance can provide a better view on blind areas of the surgical field, including the depth of the optic canal ( Fig. 2 ). The link to the video can be found at: https://youtu.be/fS2udUCPH1g .


Characterization of [3H]MK-801 binding to N-methyl-D-aspartate receptors in cultured rat cerebellar granule neurons and involvement in glutamate-mediated toxicity.
The conditions required for growth and survival of cerebellar granule neurons in vitro are known to alter the developmental regulation of NMDA receptor subunit mRNA. In the present report, we have examined the functional and pharmacological characteristics of NMDA receptors on cerebellar granule neurons at 12 days in culture (12 DIC). Under open-channel conditions in extensively washed membranes, [3H]MK-801 labeled a uniform population of sites (Kd = 3.2 +/- 0.3 nM) in a saturable manner (Bmax = 416 +/- 18 fmol/mg); however, biexponential association and dissociation kinetics indicated the possible existence of at least two NMDA receptor populations that differ in pharmacological properties. The kinetically derived equilibrium dissociation constants for the high- and low-affinity binding components were 0.56 and 771 nM, respectively. The equilibrium competition analysis of MK-801 and other channel-blocking compounds as displacers of [3H]MK-801 revealed the presence of high- and low-affinity binding sites with relative apportionments of 70% and 30%, respectively. The rank-order potency profile of competitor binding at the high-affinity site was (+)-MK-801 > TCP > dextrorphan > dextromethorphan > (+)-ketamine. When tested for the ability to protect 12 DIC cerebellar granule neurons from acute glutamate-induced toxicity, the neuroprotective rank-order potency of these compounds was MK-801 > TCP > dextrorphan > (+)-ketamine > dextromethorphan, which correlated significantly with the high-affinity competition binding profile and thus established the role of NMDA receptors in glutamate toxicity. The findings of these experiments indicate that NMDA receptors on 12 DIC cerebellar granule neurons are a heterogenous population that functionally mediate glutamate-induced neurotoxicity. The heterogenous [3H]MK-801 binding sites may represent NMDA receptor channels composed of different subunits.


Association between subclinical thyroid dysfunction and change in bone mineral density in prospective cohorts.
Subclinical hyperthyroidism (SHyper) has been associated with increased risk of hip and other fractures, but the linking mechanisms remain unclear. To investigate the association between subclinical thyroid dysfunction and bone loss. Individual participant data analysis was performed after a systematic literature search in MEDLINE/EMBASE (1946-2016). Two reviewers independently screened and selected prospective cohorts providing baseline thyroid status and serial bone mineral density (BMD) measurements. We classified thyroid status as euthyroidism (thyroid-stimulating hormone [TSH] 0.45-4.49 mIU/L), SHyper (TSH < 0.45 mIU/L) and subclinical hypothyroidism (SHypo, TSH ≥ 4.50-19.99 mIU/L) both with normal free thyroxine levels. Our primary outcome was annualized percentage BMD change (%ΔBMD) from serial dual X-ray absorptiometry scans of the femoral neck, total hip and lumbar spine, obtained from multivariable regression in a random-effects two-step approach. Amongst 5458 individuals (median age 72 years, 49.1% women) from six prospective cohorts, 451 (8.3%) had SHypo and 284 (5.2%) had SHyper. During 36 569 person-years of follow-up, those with SHyper had a greater annual bone loss at the femoral neck versus euthyroidism: %ΔBMD = -0.18 (95% CI: -0.34, -0.02; I2 = 0%), with a nonstatistically significant pattern at the total hip: %ΔBMD = -0.14 (95% CI: -0.38, 0.10; I2 = 53%), but not at the lumbar spine: %ΔBMD = 0.03 (95% CI: -0.30, 0.36; I2 = 25%); especially participants with TSH < 0.10 mIU/L showed an increased bone loss in the femoral neck (%Δ BMD = -0.59; [95% CI: -0.99, -0.19]) and total hip region (%ΔBMD = -0.46 [95% CI: -1.05, -0.13]). In contrast, SHypo was not associated with bone loss at any site. Amongst adults, SHyper was associated with increased femoral neck bone loss, potentially contributing to the increased fracture risk.


Cultural competence revisited: nursing students with disabilities.
The demographic profile of students in nursing schools is changing in relation to many different cultural backgrounds. Despite the potential for students with disabilities to enrich the nursing profession, nurse educators may be perpetuating historical attitudes, values, and practices that exclude students with disabilities from gaining admission or identifying themselves as people with disabilities. Educators in nursing schools continue to ask whether people with disabilities have a place in the nursing profession, while the more salient question is, "When will people with disabilities have a place in the nursing profession?" More important, as we create environments that welcome students with disabilities into the nursing profession, how does the quality of nursing care improve and become more appropriate for people with different cultural experiences? The purpose of this article is to present the value of recruiting students with disabilities into nursing schools in, order to enhance culturally competent nursing care.


Effects of short-chain nitrocompounds against Campylobacter jejuni and Campylobacter coli in vitro.
Effects of 2-nitro-1-propanol, 2-nitroethanol, nitroethane, and 2-nitro-methyl-propionate (0, 10, and 20 mM) on growth of Campylobacter jejuni were tested during culture in Bolton broth adjusted to pH 5.6, 7.0, or 8.2. The nitrocompounds were similarly tested against C. coli but at pH 8.2 only. Viable cell counts measured during incubation revealed main effects (P < 0.05) of all nitrocompounds on the survivability of C. jejuni. An effect of pH (P < 0.05) on the survivability of C. Jejuni during incubation with nitrocompounds was observed, with greater inhibition observed at pH 8.2 than at pH 5.6 or 7.0 for nitroethane, 2-nitro-l-propanol, and 2-nitroethanol, but not for 2-nitro-methyl-propionate, which showed greatest inhibition at pH 5.6. Except for 2-nitro-methyl-propionate, which was ineffective, all nitrocompounds elicited similar effects on C. coli. The effect of nitroethane and 2-nitro-l-propanol (10 mM) on naturally occurring Campylobacter was investigated during incubation of porcine fecal suspensions, where Campylobacter concentrations decreased more rapidly (P < 0.05) in suspensions with added 2-nitro-l-propanol than in unsupplemented or nitroethane-supplemented suspensions, thus reiterating the superior inhibitory effect of 2-nitro-l-propanol.


Results in Adult Cochlear Implant Recipients With Varied Asymmetric Hearing: A Prospective Longitudinal Study of Speech Recognition, Localization, and Participant Report.
Asymmetric hearing with severe to profound hearing loss (SPHL) in one ear and better hearing in the other requires increased listening effort and is detrimental for understanding speech in noise and sound localization. Although a cochlear implant (CI) is the only treatment that can restore hearing to an ear with SPHL, current candidacy criteria often disallows this option for patients with asymmetric hearing. The present study aimed to evaluate longitudinal performance outcomes in a relatively large group of adults with asymmetric hearing who received a CI in the poor ear. Forty-seven adults with postlingual hearing loss participated. Test materials included objective and subjective measures meant to elucidate communication challenges encountered by those with asymmetric hearing. Test intervals included preimplant and 6 and 12 months postimplant. Preimplant testing was completed in participants' everyday listening condition: bilateral hearing aids (HAs) n = 9, better ear HA n = 29, and no HA n = 9; postimplant, each ear was tested separately and in the bimodal condition. Group mean longitudinal results in the bimodal condition postimplant compared with the preimplant everyday listening condition indicated significantly improved sentence scores at soft levels and in noise, improved localization, and higher ratings of communication function by 6 months postimplant. Group mean, 6-month postimplant results were significantly better in the bimodal condition compared with either ear alone. Audibility and speech recognition for the poor ear alone improved significantly with a CI compared with preimplant. Most participants had clinically meaningful benefit on most measures. Contributory factors reported for traditional CI candidates also impacted results for this population. In general, older participants had poorer bimodal speech recognition in noise and localization abilities than younger participants. Participants with early SPHL onset had better bimodal localization than those with later SPHL onset, and participants with longer SPHL duration had poorer CI alone speech understanding in noise but not in quiet. Better ear pure-tone average (PTA) correlated with all speech recognition measures in the bimodal condition. To understand the impact of better ear hearing on bimodal performance, participants were grouped by better ear PTA: group 1 PTA ≤40 dB HL (n = 19), group 2 PTA = 41 to 55 dB HL (n = 14), and group 3 PTA = 56 to 70 dB HL (n = 14). All groups showed bimodal benefit on speech recognition measures in quiet and in noise; however, only group 3 obtained benefit when noise was toward the CI ear. All groups showed improved localization and ratings of perceived communication. Receiving a CI for the poor ear was an effective treatment for this population. Improved audibility and speech recognition were evident by 6 months postimplant. Improvements in sound localization and self-reports of communication benefit were significant and not related to better ear hearing. Participants with more hearing in the better ear (group 1) showed less bimodal benefit but greater bimodal performance for speech recognition than groups 2 and 3. Test batteries for this population should include quality of life measures, sound localization, and adaptive speech recognition measures with spatially separated noise to capture the hearing loss deficits and treatment benefits reported by this patient population.


Knocking out the glial glutamate transporter GLT-1 reduces glutamate uptake but does not affect hippocampal glutamate dynamics in early simulated ischaemia.
Glutamate release in ischaemia triggers neuronal death. The major glial glutamate transporter, GLT-1, might protect against glutamate-evoked death by removing extracellular glutamate, or contribute to death by reversing and releasing glutamate. Previous studies of the role of GLT-1 in ischaemia have often used the GLT-1 blocker dihydrokainate at concentrations that affect transporters other than GLT-1 and which affect kainate, N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors. In hippocampal slices from postnatal day 14 mice lacking GLT-1, the current response of area CA1 pyramidal cells to superfused AMPA and NMDA (which are not taken up) was unaffected, whereas the response to 100 microm glutamate was more than doubled relative to that in wild-type littermates, a finding consistent with a decrease in glutamate uptake. In response to a few minutes of simulated ischaemia, pyramidal cells in wild-type mice showed a large and sudden inward glutamate-evoked current [the anoxic depolarization (AD) current], which declined to a less inward plateau. In mice lacking GLT-1, the time to the occurrence of the AD current, its amplitude, the size of the subsequent plateau current and the block of the plateau current by glutamate receptor blockers were all indistinguishable from those in wild-type mice. We conclude that GLT-1 does not contribute significantly to glutamate release or glutamate removal from the extracellular space in early simulated ischaemia. These data are consistent with glutamate release being by reversal of neuronal transporters, and with uptake into glia being compromised by the ischaemia-evoked fall in the level of ATP needed to convert glutamate into glutamine.


Local modulation of intracellular calcium levels near a single-cell wound in human endothelial monolayers.
An endothelial cell monolayer with a single mechanically lysed cell was used as a model to examine the extent, kinetics, and nature of local calcium mobilization in the neighborhood of a wound. Individual endothelial cells from confluent monolayers were mechanically lysed with a minutien needle coupled to a micromanipulator while producing no observable mechanical trauma to the neighboring cells. Changes in calcium levels in individual cells surrounding the wound site were monitored by epifluorescence microphotometry with the calcium-sensitive fluorophore indo-1. Individual cells adjacent to the wound site showed a substantial increase in their intracellular calcium levels, almost as high as the calcium levels attained by ionophore controls. The magnitude of intracellular calcium mobilization in confluent monolayers decreased with distance from the wound site, and those cells located at a radius greater than seven cells from the wound site showed no change in their calcium levels. Thus, lysis of a single cell resulted in calcium mobilization in approximately 200 neighboring cells. The time necessary for intracellular calcium to reach maximum levels also increased with distance from the wound site. Calcium mobilization was partly intracellular and was inhibited by disrupting cell-cell coupling or by increasing gap junction resistance by heptanol. This mobilization was greatly attenuated in subconfluent endothelial monolayers, and it was not observed in fibroblasts or smooth muscle cells; furthermore, the effect was defective in monolayers intentionally contaminated with smooth muscle cells. This study examines the extent and possible mechanisms of local endothelial activation near a microscopic endothelial wound.


Excitable waves and direction-sensing in Dictyostelium discoideum: steps towards a chemotaxis model.
In recent years, there have been significant advances in our understanding of the mechanisms underlying chemically directed motility by eukaryotic cells such as Dictyostelium. In particular, the local excitation and global inhibition (LEGI) model has proven capable of providing a framework for quantitatively explaining many experiments that present Dictyostelium cells with tailored chemical stimuli and monitor their subsequent polarization. In their natural setting, cells generate their own directional signals via the detection and secretion of cyclic adenosine monophosphate (cAMP). Here, we couple the LEGI approach to an excitable medium model of the cAMP wave-field that is propagated by the cells and investigate the possibility for this class of models to enable accurate chemotaxis to the cAMP waveforms expected in vivo. Our results indicate that the ultra-sensitive version of the model does an excellent job in providing natural wave rectification, thereby providing a compelling solution to the 'back-of-the-wave paradox' during cellular aggregation.


The effects of platelet count on clot retraction and tissue plasminogen activator-induced fibrinolysis on thrombelastography.
Clot retraction and fibrinolysis may present as a decrease in amplitude on thrombelastography (TEG). The former represents normal or hyperactive platelet function, and the latter represents a fibrinolytic state. It is important to distinguish clot retraction from fibrinolysis because the treatment of each condition is different. To distinguish between these phenomena, we performed TEG with platelet-poor plasma (PPP) and platelet-rich plasma (PRP) with an increasing platelet count (range, 50-1200 x 10(9)/L) with or without abciximab. Maximum amplitude (MA) and the percentage decrease of amplitude at 30 and 60 min after MA were examined for each sample. Blood samples to which tissue plasminogen activator (tPA) was added served as positive controls for fibrinolysis. Morphological changes of clots and D-dimer levels were also examined. With higher platelet counts, the percentage decrease of amplitude after MA increased significantly at 30 and 60 min, but not in the abciximab samples. Morphological changes of clots have shown clot retraction in PRP, but not in PPP or PRP pretreated with abciximab. D-dimer levels increased only in samples to which tPA was added, but not in native PPP or PRP samples. In conclusion, we have shown that the decrease in amplitude at 30 and 60 min can be due to platelet-mediated clot retraction and can be attenuated by sample pretreatment with abciximab, which interrupts platelet-fibrin(ogen) binding.


BRAT: bisulfite-treated reads analysis tool.
We present a new, accurate and efficient tool for mapping short reads obtained from the Illumina Genome Analyzer following sodium bisulfite conversion. Our tool, BRAT, supports single and paired-end reads and handles input files containing reads and mates of different lengths. BRAT is faster, maps more unique paired-end reads and has higher accuracy than existing programs. The software package includes tools to end-trim low-quality bases of the reads and to report nucleotide counts for mapped reads on the reference genome.


Improving the economics of NASH/NAFLD treatment through the use of systems biology.
Nonalcoholic steatohepatitis (NASH) is a severe form of nonalcoholic fatty liver disease (NAFLD). We surveyed NASH therapies currently in development, and found a significant variety of targets and approaches. Evaluation and clinical testing of these targets is an expensive and time-consuming process. Systems biology approaches could enable the quantitative evaluation of the likely efficacy and safety of different targets. This motivated our review of recent systems biology studies that focus on the identification of targets and development of effective treatments for NASH. We discuss the potential broader use of genome-scale metabolic models and integrated networks in the validation of drug targets, which could facilitate more productive and efficient drug development decisions for the treatment of NASH.


Allogeneic bone marrow transplantation using unrelated donors: a pilot study of the Canadian Bone Marrow Transplant Group.
Between February 1988 and January 1990, 35 patients underwent allogeneic bone marrow transplantation (BMT) from unrelated donors using measures routinely employed for matched related donors. Median patient age was 34 years (range 2-49). Thirty-two patients had hematologic malignancies, including chronic myelogenous leukemia (CML) in 16; three patients had severe aplastic anemia. Donor-patient pairs were matched at the HLA loci tested serologically (HLA-A, -B, -DR) in 29 cases; mixed leukocyte culture results were variable but often reactive. Five patients died prior to day +28 without evidence of myeloid engraftment, and one patient developed fatal graft failure several months after initial engraftment. Acute graft-versus-host disease (GVHD) occurred in 77% (95% confidence interval [CI] 60-90%) of all patients, and GVHD contributed to the death of 10 patients. Fatal regimen-related toxicity occurred in four patients and another died due to neurologic complications of a process that resembled the hemolytic-uremic syndrome. Two acute leukemia patients relapsed, and a CML patient was found to have a localized non-Hodgkin's lymphoma at necropsy. As of 1 June 1991, 14 patients are alive and in remission at a median follow-up of 1.9 years (range 1.5-3.3); all except one have normal performance scores. The 2-year actuarial event-free survival for all patients is 40% (95% CI 24-56%). Proportional hazards analysis revealed favorable significance for female patient sex, less advanced disease status and shorter interval from diagnosis to BMT. While unrelated-donor transplants need not necessarily duplicate the results of related-donor transplants to be of benefit, the event-free survival in this series was roughly similar to that expected in the related-donor situation, with the high transplant-related mortality somewhat offset by a low recurrence rate. Further studies using unrelated donors, employing new methods of preventing transplant-related complications, are indicated.


Sequence analysis of the thyrotropin (TSH) receptor gene in congenital primary hypothyroidism associated with TSH unresponsiveness.
Congenital primary hypothyroidism due to thyrotropin (TSH) unresponsiveness is a very rare disorder and only a few cases have been documented previously. To elucidate whether structural abnormalities in the TSH receptor (TSHR) could be a primary underlying mechanism of this disorder, we analyzed nucleotide sequence of the entire coding region of the TSHR gene in three patients diagnosed with congenital primary hypothyroidism associated with TSH unresponsiveness. Diagnosis of TSH unresponsiveness was largely made based on the following criteria: (a) congenital primary hypothyroidism with autosomal recessive inheritance, (b) a nongoitrous thyroid gland in a normal position with low thyroidal radioactive iodine uptake, (c) normal in vitro TSH bioactivity or absent in vivo response to exogenous TSH, and (d) absence of thyroid autoantibodies. The TSHR cDNA was successfully obtained from RNA of peripheral mononuclear leukocytes with reverse transcription and polymerase chain reaction, and was sequenced directly. Comparison of these nucleotide sequences with the normal TSHR sequence revealed no difference in the predicted amino acid sequence with a heterozygous polymorphism in codon 601 in one patient, indicating absence of TSHR structural abnormalities in these patients. Our results indicate that congenital primary hypothyroidism associated with TSH unresponsiveness is unlikely to be due to mutations in the TSHR-structure gene.


Development and evaluation of feeding-period average dry matter intake prediction equations from a commercial feedlot database.
Intake prediction equations were developed from a commercial feedlot database consisting of 3,363 pen means collected from 3 feedlots over a 4-yr period. The objective was to predict feeding-period average DMI from variables known at the start of the feeding period, including initial BW, sex, and average DMI from d 8 to 28 of the feeding period. Equations were evaluated within the same database using bootstrapping and cross-validation techniques. Bootstrapping evaluations of equations that included initial BW and sex explained approximately 57.0 to 73.2% of the variation in observed DMI with 90.5 to 96.6% accuracy, but DMI was overpredicted by 0.21 kg/d. Accuracy and precision improved with addition to the models of average DMI from d 8 to 28 of the feeding period; these models accounted for 68.0 to 83.0% of variation in observed DMI with model accuracy between 95.2 and 99.5% and overprediction of DMI by 0.05 kg/d. Cross-validation of developed equations confirmed the robust nature of chosen variables and the decrease in prediction error as sex and average DMI from d 8 to 28 were added to models based on initial shrunk BW (SBW). Bootstrapping and cross-validation evaluations were also conducted on the NRC (1996) NE(m)-based intake prediction equations with and without the NRC-recommended 4% decrease in predicted DMI and a 12% increase in dietary NE(m) concentration associated with the feeding of monensin. The average metabolic SBW used in these equations was computed from initial SBW and a predicted final SBW developed from the pen database. When these evaluations were performed, model precision decreased compared with simpler equations that included initial BW and sex, but using both adjustments for use of monensin improved the accuracy of the NRC equation. Nonetheless, the systematic bias proportion of the mean square error of prediction for bootstrapping analyses increased from 4.6 to 11.1% in the model with no adjustments to 16.3 to 38.9% in the model with both monensin adjustments, demonstrating these adjustments are less than optimal. Overall, variables that are typically available to feedlot managers when cattle are started on feed (e.g., initial BW, sex, and an early indication of DMI) were important predictors of feeding-period average DMI by pens of cattle.


Evaluation of reference genes for RT qPCR analyses of structure-specific and hormone regulated gene expression in Physcomitrella patens gametophytes.
The use of the moss Physcomitrella patens as a model system to study plant development and physiology is rapidly expanding. The strategic position of P. patens within the green lineage between algae and vascular plants, the high efficiency with which transgenes are incorporated by homologous recombination, advantages associated with the haploid gametophyte representing the dominant phase of the P. patens life cycle, the simple structure of protonemata, leafy shoots and rhizoids that constitute the haploid gametophyte, as well as a readily accessible high-quality genome sequence make this moss a very attractive experimental system. The investigation of the genetic and hormonal control of P. patens development heavily depends on the analysis of gene expression patterns by real time quantitative PCR (RT qPCR). This technique requires well characterized sets of reference genes, which display minimal expression level variations under all analyzed conditions, for data normalization. Sets of suitable reference genes have been described for most widely used model systems including e.g. Arabidopsis thaliana, but not for P. patens. Here, we present a RT qPCR based comparison of transcript levels of 12 selected candidate reference genes in a range of gametophytic P. patens structures at different developmental stages, and in P. patens protonemata treated with hormones or hormone transport inhibitors. Analysis of these RT qPCR data using GeNorm and NormFinder software resulted in the identification of sets of P. patens reference genes suitable for gene expression analysis under all tested conditions, and suggested that the two best reference genes are sufficient for effective data normalization under each of these conditions.


Delinking CARD9 and IL-17: CARD9 Protects against Candida tropicalis Infection through a TNF-α-Dependent, IL-17-Independent Mechanism.
Candida is the third most common cause of bloodstream infections in hospitalized patients. Immunity to C. albicans, the most frequent species to be isolated in candidiasis, involves a well-characterized Dectin-1/caspase-associated recruitment domain adaptor 9 (CARD9)/IL-17 signaling axis. Infections caused by non-albicans Candida species are on the rise, but surprisingly little is known about immunity to these pathogens. In this study, we evaluated a systemic infection model of C. tropicalis, a clinically relevant, but poorly understood, non-albicans Candida. Mice lacking CARD9 were profoundly susceptible to C. tropicalis, displaying elevated fungal burdens in visceral organs and increased mortality compared with wild-type (WT) controls. Unlike C. albicans, IL-17 responses were induced normally in CARD9(-/-) mice following C. tropicalis infection. Moreover, there was no difference in susceptibility to C. tropicalis infection between WT and IL-23p19(-/-), IL-17RA(-/-), or Act1(-/-) mice. However, TNF-α expression was markedly impaired in CARD9(-/-) mice. Consistently, WT mice depleted of TNF-α were more susceptible to C. tropicalis, and CARD9-deficient neutrophils and monocytes failed to produce TNF-α following stimulation with C. tropicalis Ags. Both neutrophils and monocytes were necessary for defense against C. tropicalis, because their depletion in WT mice enhanced susceptibility to C. tropicalis. Disease in CARD9(-/-) mice was not due to defective neutrophil or monocyte recruitment to infected kidneys. However, TNF-α treatment of neutrophils in vitro enhanced their ability to kill C. tropicalis. Thus, protection against systemic C. tropicalis infection requires CARD9 and TNF-α, but not IL-17, signaling. Moreover, CARD9-dependent production of TNF-α enhances the candidacidal capacity of neutrophils, limiting fungal disease during disseminated C. tropicalis infection.


Concurrent protective and destructive signaling of JNK2 in neuroblastoma cells.
Investigation of the c-Jun N-terminal kinases (JNKs) has mainly focused on their response to stress and their pro-apoptotic effects. In this regard, JNKs are crucial mediators of chemotherapy-induced killing of tumor cells. Importantly, however, JNKs also have physiological functions in cancer involving cell cycle regulation or oncogenesis. Hypothetically, the composition of JNK signalosomes determines the signaling outcome which,in turn, implies a multitude of different, sometimes opposing and interfering functions. In the present study,the well-characterized human neuroblastoma cell line SH-SY5Y served as a model system to separate physiological and pro-apoptotic JNK actions in the response to the cytoskeleton-interfering substances colchicine, cytochalasin D and taxol. Basically, JNKs mediated both cell death and proliferation. Using the chemical JNK inhibitor SP600125 as well as compartment-specific JNK-inhibiting constructs and dominant negative isoform mutants, we show that the nuclear subgroup of JNK2 is the dominant effector in colchicine and taxol-induced apoptosis, while cell cycle promotion is mediated by both cytoplasmic and nuclear JNK2.In contrast, cytochalasin D-triggered apoptosis is independent of JNK signaling. Interestingly, the data of the present study demonstrate for the first time that both cell protective (cell cycle progression) and destructive mechanisms (apoptosis) are simultaneously controlled by a single JNK isoform in the same cell system even under the influence of one stimulus. This has implications for the therapeutic application of JNK inhibitors and cytoskeleton-interfering substances in oncologic disorders.


Anticipatory governance for social-ecological resilience.
Anticipation is increasingly central to urgent contemporary debates, from climate change to the global economic crisis. Anticipatory practices are coming to the forefront of political, organizational, and citizens' society. Research into anticipation, however, has not kept pace with public demand for insights into anticipatory practices, their risks and uses. Where research exists, it is deeply fragmented. This paper seeks to identify how anticipation is defined and understood in the literature and to explore the role of anticipatory practice to address individual, social, and global challenges. We use a resilience lens to examine these questions. We illustrate how varying forms of anticipatory governance are enhanced by multi-scale regional networks and technologies and by the agency of individuals, drawing from an empirical case study on regional water governance of Mälaren, Sweden. Finally, we discuss how an anticipatory approach can inform adaptive institutions, decision making, strategy formation, and societal resilience.


Effects of prolonged strenuous exercise (marathon running) on biochemical and haematological markers used in the investigation of patients in the emergency department.
To investigate the effects of strenuous exercise on commonly used biochemical and haematological variables in subjects running the 2002 London marathon. 34 healthy volunteers (7 female, 27 male) were recruited for the study. Blood was taken before the start (at registration) and immediately after completion of the marathon. Samples were analysed for urea and electrolytes, liver function tests, creatine kinase (CK), CK-MB isoenzyme, myoglobin, troponin I, full blood count, a clotting screen, and D-dimers. The results before and after exercise were compared. Pearson's correlation coefficients were calculated for all variables. Significant increases were found in CK, CK-MB, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and myoglobin following the marathon. However, there was no significant change in the level of troponin I. There was also evidence of activation of the coagulation and fibrinolytic cascades following the marathon, with a reduction in activated partial thromboplastin time, a reduction in fibrinogen, and an increase in D-dimers. The results confirm previous individual studies on marathon running and the biochemical and haematological tests routinely carried out in hospital. These are affected by prolonged exercise, and "abnormal" results in these tests may be normal after prolonged exercise and therefore not diagnostic of a disease process. The results of investigations in patients who have been exercising should be interpreted with caution.


Comparative genomic hybridizations reveal genetic regions within the Mycobacterium avium complex that are divergent from Mycobacterium avium subsp. paratuberculosis isolates.
Mycobacterium avium subsp. paratuberculosis is genetically similar to other members of the Mycobacterium avium complex (MAC), some of which are nonpathogenic and widespread in the environment. We have utilized an M. avium subsp. paratuberculosis whole-genome microarray representing over 95% of the predicted coding sequences to examine the genetic conservation among 10 M. avium subsp. paratuberculosis isolates, two isolates each of Mycobacterium avium subsp. silvaticum and Mycobacterium avium subsp. avium, and a single isolate each of both Mycobacterium intracellulare and Mycobacterium smegmatis. Genomic DNA from each isolate was competitively hybridized with DNA from M. avium subsp. paratuberculosis K10, and open reading frames (ORFs) were classified as present, divergent, or intermediate. None of the M. avium subsp. paratuberculosis isolates had ORFs classified as divergent. The two M. avium subsp. avium isolates had 210 and 135 divergent ORFs, while the two M. avium subsp. silvaticum isolates examined had 77 and 103 divergent ORFs. Similarly, 130 divergent ORFs were identified in M. intracellulare. A set of 97 ORFs were classified as divergent or intermediate in all of the nonparatuberculosis MAC isolates tested. Many of these ORFs are clustered together on the genome in regions with relatively low average GC content compared with the entire genome and contain mobile genetic elements. One of these regions of sequence divergence contained genes homologous to a mammalian cell entry (mce) operon. Our results indicate that closely related MAC mycobacteria can be distinguished from M. avium subsp. paratuberculosis by multiple clusters of divergent ORFs.


Service components of case management which reduce inpatient care use for persons with serious mental illness.
This is a study of two types of case management: case management (CM) which provided the service coordination functions, and Intensive Case Management (ICM) which consisted of both the coordination function and the provision of direct support to the client. Using secondary data on public clients, characteristics of mental health service use were analyzed for 80 ICM and 84 CM clients. The ICM clients had significantly fewer episodes per patient and less inpatient days per year than the CM clients. These findings suggest that direct support services make a significant difference in reducing annual hospital care.


Proton transfer in ba(3) cytochrome c oxidase from Thermus thermophilus.
The respiratory heme-copper oxidases catalyze reduction of O(2) to H(2)O, linking this process to transmembrane proton pumping. These oxidases have been classified according to the architecture, location and number of proton pathways. Most structural and functional studies to date have been performed on the A-class oxidases, which includes those that are found in the inner mitochondrial membrane and bacteria such as Rhodobacter sphaeroides and Paracoccus denitrificans (aa(3)-type oxidases in these bacteria). These oxidases pump protons with a stoichiometry of one proton per electron transferred to the catalytic site. The bacterial A-class oxidases use two proton pathways (denoted by letters D and K, respectively), for the transfer of protons to the catalytic site, and protons that are pumped across the membrane. The B-type oxidases such as, for example, the ba(3) oxidase from Thermus thermophilus, pump protons with a lower stoichiometry of 0.5 H(+)/electron and use only one proton pathway for the transfer of all protons. This pathway overlaps in space with the K pathway in the A class oxidases without showing any sequence homology though. Here, we review the functional properties of the A- and the B-class ba(3) oxidases with a focus on mechanisms of proton transfer and pumping.


Reproductive epidemiology of glial tumors may reveal novel treatments: high-dose progestins or progesterone antagonists as endocrino-immune modifiers against glioma.
Female gender, contraceptives, and menopausal hormone replacement treatments containing progesterone analogues associate with higher risk of meningiomas yet with lower risk of gliomas. Progesterone receptor (PR) expression and mifepristone treatment was highly discussed for meningiomas. However, much less is known in regard to progesterone actions in gliomas despite PR expression strongly correlates with their grade. Meningiomas and gliomas may grow faster during gestation; but paradoxically, parousity reduces lifetime risk of gliomas which can be explained with dichotomous cell growth-stimulating and inhibitory actions of progesterone at low versus high levels. Progesterone levels gradually increase in gestation up to 200-fold and the incidence of highly angiogenic brain tumors decreases in the last trimester. Indeed, progesterone stimulates glial tumor cell growth at low doses (10 nM) while induces cell kill at higher doses. During gestation, some immune pathways are activated to protect the mother and the fetus against microbial pathogens. In parallel, high-dose medroxyprogesterone acetate (MPA) used in treatment of endometrial carcinoma decreases tumoral expression of PR-B and increases infiltration of cytotoxic T lymphocytes and natural killer cells. MPA also synergies with IL-2 in clinical treatment of renal cancer. In both glioma and meningioma, the dominant cytosolic PR is PR-B which increases cell growth, while PR-A limits cell growth. This seems also paradoxical at the first glance due to opposite behavior of these tumors in diverse endocrine conditions. High-dose progestins may inhibit brain tumor growth by downregulating PR-B, yet the dosage thresholds may differ between glial and meningeal tumors due to higher total PR expression in meningiomas. Supporting this proposal, certain progestins were reported to stimulate meningioma growth in anecdotal reports, but same agents at much higher doses reduced meningioma cell proliferation in pilot clinical studies. PR antagonist mifepristone reduced meningioma growth in some clinical studies, but lacked efficacy in others. In fact, mifepristone also has partial PR agonist efficacy and acts in synergy with MPA to block EC growth. Hence, a similar mechanism of receptor downregulation may also account for mifepristone. Both MPA and mifepristone also harbor myeloprotective features against chemotherapy. Ulipristal is another contraceptive PR antagonist and exerts promising anticancer activity on drug-resistant ovarian cancer and BRCA1-mutant breast cancer cells, which can be tested in animal glioblastoma models. We propose that progestins strongly deserve to be investigated in experimental models of glioblastoma alone and in combination with immunostimulating agents.


The Donders model of the circulation in normo- and pathophysiology.
The solution of some recent as well as of long standing problems, unanswerable due to experimental inaccessibility or moral objections are addressed. In this report, a model of the closed human cardiovascular loop is developed. This model, using one set of 88 equations, allows variations from normal resting conditions to exercise, as well as to the ultimate condition of a circulation following cardiac arrest. The principal purpose of the model is to evaluate the continuum of physiological conditions to cardiopulmonary resuscitation (CPR) effects within the circulation.Within the model, Harvey's view of the circulation has been broadened to include impedance-defined flow as a unifying concept, and as a mechanism in CPR. The model shows that depth of respiration, sympathetic stimulation of cardiac contractile properties and baroreceptor activity can exert powerful influences on the increase in cardiac output, while heart and respiratory rate increases tend to exert an inhibiting influence, with the pressure and flow curves compatible with accepted references. Impedance-defined flow encompasses both positive and negative effects.The model also demonstrates the limitations to cardiopulmonary resuscitation caused by external force applied to intrathoracic structures, with effective cardiac output being limited by collapse and sloshing. Stroke volumes from 6 to 51 ml are demonstrated. It shows that the clinical inclination to apply high pressures to intrathoracic structures may not be rewarded with improved net flow.


Cancer Stem Cells and Radioresistance: Rho/ROCK Pathway Plea Attention.
Radiation is the most potent mode of cancer therapy; however, resistance to radiation therapy results in tumor relapse and subsequent fatality. The cancer stem cell (CSC), which has better DNA repair capability, has been shown to contribute to tumor resistance and is an important target for treatment. Signaling molecules such as Notch, Wnt, and DNA repair pathways regulate molecular mechanisms in CSCs; however, none of them have been translated into therapeutic targets. The RhoGTPases and their effector ROCK-signaling pathway, though important for tumor progression, have not been well studied in the context of radioresistance. There are reports that implicate RhoA in radioresistance. ROCK2 has also been shown to interact with BRCA2 in the regulation of cell division. Incidentally, statins (drug for cardiovascular ailment) are functional inhibitors of RhoGTPases. Studies suggest that patients on statins have a better prognosis in cancers. Data from our lab suggest that ROCK signaling regulates radioresistance in cervical cancer cells. Collectively, these findings suggest that Rho/ROCK signaling may be important for radiation resistance. In this review, we enumerate the role of Rho/ROCK signaling in stemness and radioresistance and highlight the need to explore these molecules for a better understanding of radioresistance and development of therapeutics.


Complete Genome Sequence of Spiroplasma kunkelii Strain CR2-3x, Causal Agent of Corn Stunt Disease in Zea mays L.
Spiroplasma kunkelii causes corn stunt disease of Zea mays L. in the Americas. Here, we report the nucleotide sequence of the 1,463,926-bp circular chromosome and four plasmids of strain CR2-3x. This information will facilitate studies of Spiroplasma pathogenicity and evolutionary adaptations to transkingdom parasitism in plants and insect vectors.


Frequent expression of MDR1 and MDR3 genes in acute myelocytic leukemia cells with t(8;21) (q22;q22).
Multidrug resistance (MDR) 1 and MDR3 gene expression were examined in acute myelocytic leukemia (AML) cells from 126 Japanese patients. In 119 AML patients at diagnosis 52 were revealed to have P-gp/MDR1 (43.7%). AML cases with t(8;21) had a higher incidence of P-gp expression (13/17; 76.5%) than cases with other karyotypes (33/89; 37.1%) (p=0.0062). CD19(+) cases expressed P-gp frequently (17/25) as did CD7(+) cases (24/35). In 17 CD19(+) AML with P-gp expression, 12 had t(8;21) abnormality. In 68 AML samples examined, MDR3 mRNA was detected in 11 cases, 9 of which had the t(8;21) abnormality. The MDR3(+) t(8;21) AML samples were also positive for CD19. We analyzed P-gp expression at both diagnosis and relapse in 18 AML patients. All 11 P-gp(+) cases at relapse, in which 5 patients were P-gp negative at diagnosis, showed either t(8;21) or CD7 positivity. Our data demonstrated that expression of MDR1 and MDR3 in AML is closely associated with chromosome abnormality t(8;21) and expression of immature lymphoid antigen CD19 as well as CD7. Kasumi-1, a t(8;21) AML cell line, was demonstrated to lose P-gp by the treatment of 1,25(OH)(2)D-3, a monocyte/macropharge differentiation inducer, suggesting the possible contribution to the therapy for AML.


Effect of intravitreal methotrexate and aqueous humor cytokine levels in refractory retinal vasculitis in Behcet disease.
The purpose of this study was to demonstrate the effect of intravitreal methotrexate (MTX) in the treatment of refractory retinal vasculitis due to Behcet disease and to evaluate the changes in aqueous humor cytokines after intravitreal MTX. Seven eyes of seven patients with Behcet disease were assessed. Patients had unilaterally active retinal vasculitis that failed to respond to conventional treatment, together with a history of increased intraocular pressure or intolerance to corticosteroid administration. Intravitreal injection of 400 μg MTX was given monthly until visual acuity and intraocular inflammation were stable. Aqueous humor samples were obtained from the patients before MTX injection and from nine eyes during cataract surgery as a control group. Changes in visual acuity with the Early Treatment Diabetic Retinopathy Study Chart, fluorescein angiographic leakage, and ocular complications associated with intravitreal injection were analyzed. Aqueous humor cytokine levels were measured by proteomic analysis using a multianalyte biochip array system. Visual acuity improved significantly during a mean follow-up of 24.9 ± 8.2 weeks with a mean improvement from baseline visual acuity of 3.3 ± 2.6 lines after a mean of 4.3 ± 1.0 injections. Six patients (85.7%) showed an increase in visual acuity by 3 or more lines, and 4 patients exhibited a decrease in fluorescein leakage. Change in intraocular pressure after intravitreal MTX was not significant. The aqueous humor levels of interleukin (IL) -6, IL-8, vascular endothelial growth factor, and monocyte chemotactic protein 1 were significantly higher in patients with Behcet disease compared with controls. The levels of IL-6 and IL-8 were significantly reduced at 4 weeks after intravitreal MTX, whereas those of vascular endothelial growth factor and monocyte chemotactic protein 1 were not reduced. Increased intraocular levels of IL-6, IL-8, vascular endothelial growth factor, and monocyte chemotactic protein 1 may be responsible for refractory retinal vasculitis in Behcet disease. Intravitreal MTX was associated with a significant reduction of IL-6 and IL-8 levels and was effective and well tolerated even in steroid responders with refractory retinal vasculitis due to Behcet disease.


Children with reported histories of sexual abuse: utilizing multiple perspectives to understand clinical and psychosocial profiles.
The current study examines multiple empirically based perspectives (i.e., child, caregiver, and clinician) of behavior and functioning as they contribute to the clinical and psychosocial profile of children (aged 5 to 17.5 years) with reported histories of sexual abuse. A large, multi-site data set of children referred into Comprehensive Community Mental Health Services both with and without reported histories of sexual abuse, was examined. Seven hundred and fifty-nine children with a reported history of sexual abuse were compared to 2722 without such a history on caregiver and child reported behavior, clinician rated functioning, diagnosis, demographic variables, and life challenges. The multiple perspectives contributed unique and specific information to regression models: caregiver-reported behavior contributed information about externalizing behavior while child-reported behavior added information about internalizing behavior and clinician ratings about self-harmful behavior. Children with reported histories of sexual abuse were also more likely to be female, Caucasian, and have reported histories of life challenges (e.g., physical abuse, substance use, running away). Child sexual abuse was associated with higher rates of depression and anxiety diagnoses, and lower rates of substance abuse, conduct, and attention deficit disorder diagnoses. The findings indicate that the profile of children entering into Comprehensive Community Mental Health Services with reported histories of sexual abuse, as compared to those without such histories, is complex and best understood via multiple perspectives. Caregiver, child and clinician rated information, when taken together, provide a comprehensive clinical and psychosocial profile around which to plan and implement individualized service plans.


Cortical Correlates of Binaural Temporal Processing Deficits in Older Adults.
This study was designed to evaluate binaural temporal processing in young and older adults using a binaural masking level difference (BMLD) paradigm. Using behavioral and electrophysiological measures within the same listeners, a series of stimulus manipulations was used to evaluate the relative contribution of binaural temporal fine-structure and temporal envelope cues. We evaluated the hypotheses that age-related declines in the BMLD task would be more strongly associated with temporal fine-structure than envelope cues and that age-related declines in behavioral measures would be correlated with cortical auditory evoked potential (CAEP) measures. Thirty adults participated in the study, including 10 young normal-hearing, 10 older normal-hearing, and 10 older hearing-impaired adults with bilaterally symmetric, mild-to-moderate sensorineural hearing loss. Behavioral and CAEP thresholds were measured for diotic (So) and dichotic (Sπ) tonal signals presented in continuous diotic (No) narrowband noise (50-Hz wide) maskers. Temporal envelope cues were manipulated by using two different narrowband maskers; Gaussian noise (GN) with robust envelope fluctuations and low-noise noise (LNN) with minimal envelope fluctuations. The potential to use temporal fine-structure cues was controlled by varying the signal frequency (500 or 4000 Hz), thereby relying on the natural decline in phase-locking with increasing frequency. Behavioral and CAEP thresholds were similar across groups for diotic conditions, while the masking release in dichotic conditions was larger for younger than for older participants. Across all participants, BMLDs were larger for GN than LNN and for 500-Hz than for 4000-Hz conditions, where envelope and fine-structure cues were most salient, respectively. Specific age-related differences were demonstrated for 500-Hz dichotic conditions in GN and LNN, reflecting reduced binaural temporal fine-structure coding. No significant age effects were observed for 4000-Hz dichotic conditions, consistent with similar use of binaural temporal envelope cues across age in these conditions. For all groups, thresholds and derived BMLD values obtained using the behavioral and CAEP methods were strongly correlated, supporting the notion that CAEP measures may be useful as an objective index of age-related changes in binaural temporal processing. These results demonstrate an age-related decline in the processing of binaural temporal fine-structure cues with preserved temporal envelope coding that was similar with and without mild-to-moderate peripheral hearing loss. Such age-related changes can be reliably indexed by both behavioral and CAEP measures in young and older adults.


Reasons behind the rising rate of involuntary admissions under the Mental Health Act (1983): Service use and cost impact.
There has been a significant rise in the use of the Mental Health Act (1983) in England over the last 10 years. This includes both health-based Place of Safety detentions and involuntary admissions to NHS mental health facilities. Although these trends should clearly inform the implementation of mental health care and legislation, there is currently little understanding of what caused these increased rates. We therefore sought to explore potential underlying reasons for the increase in involuntary admissions and Place of Safety detentions and to ascertain the associated service costs. We extracted publicly available data to ascertain the observed number of involuntary admissions (Section 2 or 3) and health-based Place of Safety detentions in England between 1999/2000 and 2015/2016. A simple regression analysis then enabled us to compare observed admission rates with predicted rates, between 2008/2009 and 2015/2016. This prediction model was based on observed figures before 2008. We then generated a costing model for these rates and compared admission costs to alternative interventions. Finally, we added relevant covariates to the prediction model, to explore potential relationships with observed rates. Since 2008/2009, there has been a marked increase in the number of involuntary admissions (38%) and Place of Safety detentions (617%). The analysis revealed that for involuntary admissions, the period of greatest increase occurred after 2012, two years after austerity measures were implemented. For Place of Safety detentions, substantial rises were seen from 2008/2009 to 2015/2016, coinciding with the economic recession. The rise in Place of Safety detentions may have been worsened by a reduction in mental health bed availability. During the study period, involuntary admissions are estimated to have cost the English NHS £6.8 billion; with a further £120 million spent on Place of Safety detentions. This is approximately £597 million greater than predicted, had involuntary admissions continued to change at pre-2008 rates. We conclude that the rise in involuntary admissions, and to a lesser extent Place of Safety detentions, were associated with three specific impactful events: the economic recession, legislative changes and the impact of austerity measures on health and social care services. In addition to the extensive arguments presented elsewhere, there is also an urgent economic case for addressing this trend.


Molecular analysis of Plasmodium falciparum recrudescent malaria infections in children treated with chloroquine in Nigeria.
Parasite genotyping by a polymerase chain reaction was used to distinguish recrudescent from newly acquired Plasmodium falciparum infections in 50 of 160 Nigerian children taking part in a chloroquine efficacy study in Ibadan, Nigeria. A finger prick blood sample was taken from each child before and after treatment to identify recrudescent parasites. By investigating allelic variation in three polymorphic antigen loci, merozoite surface protein-1 (MSP-1), MSP-2, and glutamate-rich protein (GLURP), we determined parasite diversity in the population and in the infected host. DNA from pretreatment and post-treatment samples from 47 of the 50 patients who failed therapy was successfully amplified by the PCR. The MSP-1, MSP-2, and GLURP genotypes in all samples showed extensive diversity, indicating polyclonal infections. The average number of clones per infection in pre-treatment sample was 2.5 with MSP-1, 4.9 with MSP-2, and 2 with GLURP. The extent of multiplicity decreased significantly (P = 0.016) in posttreatment samples. Multiplicity of infection and initial parasite density were not age dependent. Comparison of the variant alleles in pretreatment and post-treatment samples of each patient indicates that 26 of the 47 children had genuinely recrudescent disease. Conversely, post-treatment samples from five children showed completely new genotypes, indicating either a previously sequestered population of parasites or a newly acquired infection. Overall, this study has shown the diversity and complexity of P. falciparum population in Ibadan, Nigeria. The study has also shown the dynamics of P. falciparum infections in this population before and after chloroquine treatment in an area of high malaria transmission.


Scalp topography of the spontaneous K-complex and of delta-waves in human sleep.
Together with spindles, K-complexes are well known hallmarks of stage 2 sleep (S2). However, little is known about their topographical distribution in comparison to delta-waves and to K-complexes superimposed by spindles. In this study, the topographical distribution of spontaneous K-complexes and delta-waves in S2 and delta-waves in stage 4 sleep (S4) in 10 healthy young adults (aged 20 to 35 years, 7 female) was investigated. K-complexes with and without spindles in S2, delta-waves with and without spindles in S2, and delta-waves in S4 distributed all over the night were visually selected. EEG power maps and statistical parametric maps were calculated. Absolute delta power of S2 K-complexes appeared to be significantly higher than of S2 delta-waves and delta power of S4 delta-waves was higher than of S2 delta-waves. In K-complexes and delta-waves, power was found to be highest over medio-frontal regions in the delta frequency band (0.5-4.0 Hz) with a second maximum occipitally in delta-waves, no matter whether superimposed by a spindle or not. K-complexes and delta-waves in S2 differ in topographical distribution. Even though in S2 delta-waves have less power, they have a similar topographical distribution in S2 and S4, supporting the hypothesis that delta-waves in S2 further develop towards delta-waves in slow wave sleep. The delta frequency components of K-complexes and delta-waves are unaffected by spindles.


The expression of the growth associated protein B50/GAP43 in the olfactory system of neonatal and adult rats.
B50/GAP43 is a neuron-specific phosphoprotein whose expression is associated with neural development and synaptic plasticity. Its postnatal ontogeny was investigated in the primary olfactory pathway of the rat using immunohistochemical methods. The unique ability of the olfactory neuroepithelium to generate new neurons from a population of precursor cells present in the basal cell layer of this tissue makes it a valuable model in the study of neural development. In newborn rats B50/GAP43 is present throughout the entire population of olfactory receptor neurons. These cells are stained throughout, from the ciliated dendritic knob to their axon terminals in the bulb. This appears to be the first example of unambiguous B50/GAP43 expression in dendritic processes. With increasing age the distribution of this protein becomes progressively restricted to a subpopulation of olfactory neurons. Comparison of the expression of B50/GAP43 and the olfactory marker protein (OMP), a polypeptide only present in mature olfactory neurons, revealed that during postnatal development of the olfactory system these 2 proteins are expressed in a nearly reciprocal fashion. In adult animals (3.5 months-6 months of age), B50/GAP43-positive cells are exclusively present adjacent to the basal cell layer of the neuroepithelium. Basal cells appear to be unstained. The region of the epithelium containing the B50/GAP43-positive cells is virtually devoid of OMP-positive neurons. A significant fraction of these B50/GAP43-containing cells bear dendritic and neuritic processes. However, these cells do not express olfactory cilia. It is probable that the olfactory neurons expressing the growth-associated B50/GAP43 protein may correspond to a particular subset of olfactory neurons at an intermediate state of maturation.


Progression in substance use initiation: A multilevel discordant monozygotic twin design.
Considerable attention has been paid to the "gateway" pattern of drug use initiation in which individuals progress from tobacco and alcohol use to cannabis and other illicit drugs. The extent to which this sequence reflects a causal impact of licit substance use on illicit substance involvement remains unclear. Clarifying the mechanisms underlying substance use initiation may help inform our understanding of risk for psychopathology, as increasing research is demonstrating associations between initiation patterns and heavier involvement. This study examined patterns of substance use initiation using a discordant twin design. Participants were 3,476 monozygotic twins (37% male) from the Australian Twin Registry who reported on their ages of tobacco, alcohol, and cannabis initiation. Multilevel proportional hazard regression models were used to (a) estimate within-twin-pair and between-twin-pair contributions to associations between the ages of onset of different drugs; and (b) examine whether the magnitude of effects differed as a function of the order of substance use initiation. Finding significant effects within twin pairs would support the hypothesis that the age of initiation of a substance causally influences the age of initiation of a subsequent substance. Finding significant effects between twin pairs would support the operation of familial influences that explain variation in the ages of initiation of multiple drugs. Within-twin-pair effects for typical patterns were modest. When initiation was atypical, however, larger within-twin-pair effects were observed and causal influences were more strongly implicated. Results support the utility of examining the timing and ordering of substance use initiation within sophisticated, genetically informative designs.


Effects of word form on brain processing of written Chinese.
Both logographic characters and alphabetic pinyins can be used to write words in Chinese. Here we use fMRI to address the question of whether the written form affects brain processing of a word. Fifteen healthy, right-handed, native Chinese-reading volunteers participated in our study and were asked to read silently either Chinese characters (8 subjects) or pinyins (7 subjects). The stimulus presentation rate was varied for both tasks to allow us to identify brain regions with word-load-dependent activation. Rate effects (fast minus slow presentations) for Chinese character reading were observed in striate and extrastriate visual cortex, superior parietal lobule, left posterior middle temporal gyrus, bilateral inferior temporal gyri, and bilateral superior frontal gyri. Rate effects for pinyin reading were observed in bilateral fusiform, lingual, and middle occipital gyri, bilateral superior parietal lobule/precuneus, left inferior parietal lobule, bilateral inferior temporal gyrus, left middle temporal gyrus, and left superior temporal gyrus. These results demonstrate that common regions of the brain are involved in reading both Chinese characters and pinyins, activated apparently independently of the surface form of the word. There also appear to be brain regions in which activation is dependent on word form. However, it is unlikely that these are entirely specific for a given word form; their activation more likely reflects relative functional specializations within broader networks for processing written language.


Invasion genetics of the chestnut blight fungus Cryphonectria parasitica in Switzerland.
Cryphonectria parasitica is the best-known example of an invasive forest pathogen in Europe. In southern Switzerland, chestnut blight was first reported in 1948 whereas, north of the Alps, it did not appear until the 1980s. Between 1995 and 2008, we sampled 640 C. parasitica isolates from nine populations south of the Alps and nine north of the Alps. Twelve historical isolates, collected between 1950 and 1972 in the south, were obtained from our collection. All 652 isolates were screened at 10 microsatellite loci to test for the existence of divergent genetic pools and to infer possible origins of haplotypes. In total, 52 haplotypes were identified. Structure software analysis indicated that 43 haplotypes (including all historical haplotypes) belonged to a main cluster, 6 haplotypes belonged to a different cluster, and 3 haplotypes had an intermediate allele pattern. All newly founded populations in northern Switzerland were initiated by one or just a few haplotypes from the main cluster, which probably came directly from the populations south of the Alps. Subsequently, genetic diversity increased through mutations, sexual reproduction, or new migrations. The highest increase in diversity was observed in populations where haplotypes from different genetic pools were encountered.


Topographical organization of the pedunculopontine nucleus.
Neurons in the pedunculopontine nucleus (PPN) exhibit a wide heterogeneity in terms of their neurochemical nature, their discharge properties, and their connectivity. Such characteristics are reflected in their functional properties and the behaviors in which they are involved, ranging from motor to cognitive functions, and the regulation of brain states. A clue to understand this functional versatility arises from the internal organization of the PPN. Thus, two main areas of the PPN have been described, the rostral and the caudal, which display remarkable differences in terms of the distribution of neurons with similar phenotype and the projections that originate from them. Here we review these differences with the premise that in order to understand the function of the PPN it is necessary to understand its intricate connectivity. We support the case that the PPN should not be considered as a homogeneous structure and conclude that the differences between rostral and caudal PPN, along with their intrinsic connectivity, may underlie the basis of its complexity.